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Medium Chain Length Polyhydroxyalkanoate Production by Engineered Pseudomonas gessardii Using Acetate-formate as Carbon Sources: Woo Young Kim, Seung-Jin Kim, Hye-Rin Seo, Yoonyong Yang, Jong Seok Lee, Moonsuk Hur, Byoung-Hee Lee, Jong-Geol Kim, Min-Kyu Oh; J. Microbiol. 2024;62(7):569-579. Published online May 3, 2024; DOI: https://doi.org/10.1007/s12275-024-00136-x

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Production of medium chain length polyhydroxyalkanoate (mcl-PHA) was attempted using Pseudomonas gessardii NIBRBAC000509957, which was isolated from Sunchang, Jeollabuk-do, Republic of Korea (35°24'27.7"N, 127°09'13.0"E) and effectively utilized acetate and formate as carbon sources. We first evaluated the utilization of acetate as a carbon source, revealing optimal growth at 5 g/L acetate. Then, formate was supplied to the acetate minimal medium as a carbon source to enhance cell growth. After overexpressing the acetate and formate assimilation pathway enzymes, this strain grew at a significantly higher rate in the medium. As this strain naturally produces PHA, it was further engineered metabolically to enhance mcl-PHA production. The engineered strain produced 0.40 g/L of mcl-PHA with a biomass content of 30.43% in fed-batch fermentation. Overall, this strain can be further developed to convert acetate and formate into valuable products.

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Formate-driven photoautotrophic growth and biopolymer storage in anaerobic purple bacteria
Mohammad Adib Ghazali Abdul Rahman, Bronwyn Laycock, Steven Pratt, Damien J. Batstone
Bioresource Technology.2025; 434: 132753. CrossRef
Sulphide and oleic acid synergism in accelerating mcl-PHA biopolymer production in Pseudomonas aeruginosa MCC 5300 by modulating electron transport system
Raghavendra Paduvari, Divyashree Somashekara
Biochemistry and Biophysics Reports.2025; 44: 102286. CrossRef
Unlocking efficient polyhydroxyalkanoate production by Gram-positive Priestia megaterium using waste-derived feedstocks
Xinyi Bai, Libo Xu, Kang Li, Guangbao Zhang, Mengjun Zhang, Yi Huang
Microbial Cell Factories.2025;[Epub] CrossRef
Selective utilization of formic acid and acetic acid in succinic acid fermentation broth to produce single-cell protein using Rhodotorula glutinis
Fuqiang Liu, Pengfei Wu, Lin Yu, Zitu Lü, Xinying Sun, Jiaxin Li, Lei Liu, Jing Wu, Jianan Zhang
Bioprocess and Biosystems Engineering.2025;[Epub] CrossRef

Biosynthesis of Chryseno[2,1,c]oxepin‑12‑Carboxylic Acid from Glycyrrhizic Acid in Aspergillus terreus TMZ05‑2, and Analysis of Its Anti‑inflammatory Activity: Liangliang Chen , Lin Zhao , Ju Han , Ping Xiao , Mingzhe Zhao , Sen Zhang , Jinao Duan; J. Microbiol. 2024;62(2):113-124. Published online February 27, 2024; DOI: https://doi.org/10.1007/s12275-024-00105-4

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Glycyrrhizic acid, glycyrrhetinic acid, and their oxo, ester, lactone, and other derivatives, are known for their anti-inflammatory, anti-oxidant, and hypoglycemic pharmacological activities. In this study, chryseno[2,1-c]oxepin-12-carboxylic acid (MG) was first biosynthesized from glycyrrhizic acid through sequential hydrolysis, oxidation, and esterification using Aspergillus terreus TMZ05-2, providing a novel in vitro biosynthetic pathway for glycyrrhizic acid derivatives. Assessing the influence of fermentation conditions and variation of strains during culture under stress-induction strategies enhanced the final molar yield to 88.3% (5 g/L glycyrrhizic acid). CCK8 assays showed no cytotoxicity and good cell proliferation, and anti-inflammatory experiments demonstrated strong inhibition of NO release (36.3%, low-dose MG vs. model), transcriptional downregulation of classical effective cellular factors tumor necrosis factor-α (TNF-α; 72.2%, low-dose MG vs. model), interleukin-6 (IL-6; 58.3%, low-dose MG vs. model) and interleukin-1β (IL-1β; 76.4%, low-dose MG vs. model), and decreased abundance of P-IKK-α, P-IKB-α, and P-P65 proteins, thereby alleviating inflammatory responses through the NF-κB pathway in LPS-induced RAW264.7 cells. The findings provide a reference for the biosynthesis of lactone compounds from medicinal plants.

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Effect of different crosslinking agents on carboxymethyl chitosan-glycyrrhizic acid hydrogel: Characterization and biological activities comparison
Yinbing Wu, Zimin Gu, Tingting Chen, Duntao Zu, Yuhui Gan, Honglin Chen, Jianni Yang, Xin Yu, Huaihong Cai, Pinghua Sun, Jianying Ning, Haibo Zhou, Junxia Zheng
International Journal of Biological Macromolecules.2025; 298: 139977. CrossRef
New oxepin and dihydrobenzofuran derivatives from Bauhinia saccocalyx roots and their anti-inflammatory, cytotoxic, and antioxidant activities
Lueacha Tabtimmai, Thanyathon Phonchan, Natrinee Thongprik, Sutin Kaennakam, Nuttapon Yodsin, Kiattawee Choowongkomon, Chanikan Sonklin, Supachai Jadsadajerm, Awat Wisetsai
Journal of Natural Medicines.2025; 79(3): 543. CrossRef
Efficient directional biosynthesis of isoquercitrin from quercetin by Bacillus subtilis CD-2 and its anti-inflammatory activity
Ju Han, Jingru Ma, Ruiqi He, Fan Yang, Jingyi Meng, Jiaqi Liu, Fanxing Shi, Jinao Duan, Liangliang Chen, Sen Zhang
Natural Product Research.2024; : 1. CrossRef

Silver Nanoparticles Modified with Polygonatum sibiricum Polysaccharide Improve Biocompatibility and Infected Wound Bacteriostasis: Ruonan Wang , Rongyu Li , Peng Zheng , Zicheng Yang , Cheng Qian , Zhou Wang , Senhe Qian; J. Microbiol. 2023;61(5):543-558. Published online April 13, 2023; DOI: https://doi.org/10.1007/s12275-023-00042-8

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Silver nanoparticles (AgNPs) exhibit strong antibacterial activity and do not easily induce drug resistance; however, the poor stability and biocompatibility in solution limit their widespread application. In this study, AgNPs were modified with Polygonatum sibiricum Polysaccharide (PSP) to synthesize PSP@AgNPs with good stability, biocompatibility, and antibacterial activity. When PSP@AgNP synthesis was performed under a reaction time of 70 min, a reaction temperature of 35 °C, and an AgNO3- to-PSP volume ratio of 1:1, the synthesized PSP@AgNPs were more regular and uniform than AgNPs, and their particle size was around 10 nm. PSP@AgNPs exhibited lower cytotoxicity and hemolysis, and stronger bacteriostatic activity. PSP@AgNPs damage the integrity and internal structure of cells, resulting in the leakage of intracellular nucleic acids and proteins. The rate of cell membrane damage in Escherichia coli and Staphylococcus aureus treated with PSP@ AgNPs increased by 38.52% and 43.75%, respectively, compared with that of AgNPs. PSP@AgNPs inhibit the activities of key enzymes related to antioxidant, energy and substance metabolism in cells. The inhibitory effects on the activities of superoxide dismutase (SOD), catalase (CAT), adenosine triphosphate enzyme (ATPase), malate dehydrogenase (MDH), and succinate dehydrogenase (SDH) in E. coli and S. aureus cells were significantly higher than those of AgNPs. In addition, compared with AgNPs, PSP@AgNPs promote faster healing of infected wounds. Therefore, PSP@AgNPs represent potential antibacterial agents against wound infections.

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Fabrication of Highly Stable Polyurushiol-Decorated Silver Nanoparticles and Evaluation of Their Antibacterial and Anti-Microalgae Activities
Lu Zheng, Jide Zhu, Jipeng Chen, Yanlian Xu, Lilong Jiang
Journal of Inorganic and Organometallic Polymers and Materials.2025; 35(1): 570. CrossRef
Preparation, Structure, Function, and Application of Dietary Polysaccharides from Polygonatum sibiricum in the Food Industry: A Review
Peilin Li, Huimin Yao, Hao Yue, Jiali Huang, Qi Wang, Chuanbo Ding, Lina Ma, Xinglong Liu, Min Yang
Molecules.2025; 30(5): 1098. CrossRef
Transcriptomic and metabolomic analyses of the antimicrobial activity of phenoxyethanol against phylotype IA1 and II Cutibacterium acnes
Li Shao, Lizhuang Wang, Yan Li, Laiji Ma, Hong Jiang, Fei Liu, Shanshan Jiang, Tianming Bai, Suzhen Yang
Journal of Applied Microbiology.2025;[Epub] CrossRef
Extraction, purification, structural characterization, biological activity, structure-activity relationship, and applications of polysaccharides derived from Polygonatum sibiricum: A review
Jiasu Wu, Chang Tan, Hongye Li, Siya Wang, Xingkui Wang, Shan Wang, Chong Ning, Weixuan Li, Chongting Guo
Trends in Food Science & Technology.2025; 161: 105038. CrossRef
Study on the structural characterization and biological activities of polysaccharides from Dictyophora rubrovolvata and its silver nanoparticles
Xiaonan Du, Junhui Wang, Li Gao, Jishun Zheng, Lei Zhang
International Journal of Biological Macromolecules.2025; 328: 147632. CrossRef
Elucidating the effects of nine-steaming-nine-drying on the morphology, metabolomic profile, and active ingredients of Polygonatum sibiricum Redouté
Rui Wang, Ping Li, Siyao Chen, Jinchao Guo, Yongling Liu, Huizhen Shen, Siyu Wang, Haiyan Li
Frontiers in Food Science and Technology.2025;[Epub] CrossRef
Improving the biocompatibility and antibacterial efficacy of silver nanoparticles functionalized with (LLRR)3 antimicrobial peptide
Rongyu Li, Jiaqing Mao, Peng Zheng, Ruonan Wang, Zicheng Yang, Senhe Qian
World Journal of Microbiology and Biotechnology.2024;[Epub] CrossRef
Advancing engineered approaches for sustainable wound regeneration and repair: Harnessing the potential of green synthesized silver nanoparticles
J. Nandhini, E. Karthikeyan, E. Elizabeth Rani, V.S. Karthikha, D. Sakthi Sanjana, H. Jeevitha, S. Rajeshkumar, Vijayan Venugopal, A. Priyadharshan
Engineered Regeneration.2024; 5(3): 306. CrossRef
Effect of Polygonatum sibiricum on biological toxicity of zinc oxide nanoparticles during respiratory exposure
Jingjing Yao, Wanqing Yang, Liang Tang, Dicheng Yang, Yan Xu, Shenmin Zhu, Jun Zhu
RSC Advances.2024; 14(43): 31360. CrossRef
Enhancing Healing of Infected Wounds with Glycerin‐Modified Sodium Alginate/Silk Sericin Composite Film Functionalized with Polygonatum sibiricum Polysaccharide‐Capped Silver Nanoparticles
Zicheng Yang, Rongyu Li, Ruonan Wang, Senhe Qian
ChemistrySelect.2024;[Epub] CrossRef
Host Defense Peptides: Exploiting an Innate Immune Component Against Infectious Diseases and Cancer
Taiwo Scholes Adewole, Oladiran Boniface Oladokun, Adenike Kuku
International Journal of Peptide Research and Therapeutics.2024;[Epub] CrossRef
Research progress on medicinal components and pharmacological activities of polygonatum sibiricum
Ruilian Liu, Xili Zhang, Yuhan Cai, Shuang Xu, Qian Xu, Chengli Ling, Xin Li, Wenjiao Li, Pingan Liu, Wenlong Liu
Journal of Ethnopharmacology.2024; 328: 118024. CrossRef
A comprehensive review on the potential applications of medicine Polygonatum species in the food sector
Mi Li, Bingzong Xie, Lewen Li, Yunge Zhang, Qingmin Chen, Jian Ju, Yanli Ma
Food Bioscience.2024; 60: 104116. CrossRef
Metallic elements combine with herbal compounds upload in microneedles to promote wound healing: a review
Xiao Tang, Li Li, Gehang You, Xinyi Li, Jian Kang
Frontiers in Bioengineering and Biotechnology.2023;[Epub] CrossRef

Reviews

Envelope‑Stress Sensing Mechanism of Rcs and Cpx Signaling Pathways in Gram‑Negative Bacteria: Seung-Hyun Cho , Kilian Dekoninck , Jean-Francois Collet; J. Microbiol. 2023;61(3):317-329. Published online March 9, 2023; DOI: https://doi.org/10.1007/s12275-023-00030-y

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Abstract PDF

The global public health burden of bacterial antimicrobial resistance (AMR) is intensified by Gram-negative bacteria, which have an additional membrane, the outer membrane (OM), outside of the peptidoglycan (PG) cell wall. Bacterial twocomponent systems (TCSs) aid in maintaining envelope integrity through a phosphorylation cascade by controlling gene expression through sensor kinases and response regulators. In Escherichia coli, the major TCSs defending cells from envelope stress and adaptation are Rcs and Cpx, which are aided by OM lipoproteins RcsF and NlpE as sensors, respectively. In this review, we focus on these two OM sensors. β-Barrel assembly machinery (BAM) inserts transmembrane OM proteins (OMPs) into the OM. BAM co-assembles RcsF, the Rcs sensor, with OMPs, forming the RcsF-OMP complex. Researchers have presented two models for stress sensing in the Rcs pathway. The first model suggests that LPS perturbation stress disassembles the RcsF-OMP complex, freeing RcsF to activate Rcs. The second model proposes that BAM cannot assemble RcsF into OMPs when the OM or PG is under specific stresses, and thus, the unassembled RcsF activates Rcs. These two models may not be mutually exclusive. Here, we evaluate these two models critically in order to elucidate the stress sensing mechanism. NlpE, the Cpx sensor, has an N-terminal (NTD) and a C-terminal domain (CTD). A defect in lipoprotein trafficking
results
in NlpE retention in the inner membrane, provoking the Cpx response. Signaling requires the NlpE NTD, but not the NlpE CTD; however, OM-anchored NlpE senses adherence to a hydrophobic surface, with the NlpE CTD playing a key role in this function.

Citations

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Metal-based antimicrobial agents in wound Dressings: Infection management and the challenge of antibiotic resistance
Haoyang Peng, Deqiao Dong, Shiquan Feng, Yueping Guo, Jiaqi Yu, Changran Gan, Xue Hu, Zhenmao Qin, Yan Liu, Yanan Gao
Chemical Engineering Journal.2025; 507: 160726. CrossRef
Bacterial cell wall synthesis and recycling: new antimicrobial targets and vaccine development
Jihyeon Min, Bitnara Kim, Yerim Park, Yongjun Son, Woojun Park
Critical Reviews in Microbiology.2025; : 1. CrossRef
Nitazoxanide inhibits pili assembly by targeting BamB to synergize with polymyxin B against drug-resistant Escherichia coli
Wenwen Li, Bingjie Ji, Boyu Li, Minghui Du, Linwei Wang, Jiale Tuo, Hongmei Zhou, Jian Gong, Yongshan Zhao
Biochimie.2025; 233: 47. CrossRef
Complex interplay between gene deletions and the environment uncovers cellular roles for genes of unknown function in Escherichia coli
Kaat Sondervorst, Kristina Nesporova, Matthew Herdman, Bart Steemans, Joëlle Rosseels, Sander K. Govers, Julia Willett
mSystems.2025;[Epub] CrossRef
Genome-wide characterization of hypothiocyanite stress response in Escherichia coli
Julia D. Meredith, Michael J. Gray, Melissa M. Kendall
Journal of Bacteriology.2025;[Epub] CrossRef
Terminal complement complexes with or without C9 potentiate antimicrobial activity against Neisseria gonorrhoeae
Evan R. Lamb, Alison K. Criss, Mariagrazia Pizza
mBio.2025;[Epub] CrossRef
RcsB and H-NS Both Contribute to the Repression the Expression of the csgDEFG Operon
Hiroshi Ogasawara, Azusa Tomioka, Yuki Kato
Microorganisms.2025; 13(8): 1829. CrossRef
Transcriptome reveals the role of the htpG gene in mediating antibiotic resistance through cell envelope modulation in Vibrio mimicus SCCF01
Zhenyang Qin, Kun Peng, Yang Feng, Yilin Wang, Bowen Huang, Ziqi Tian, Ping Ouyang, Xiaoli Huang, Defang Chen, Weimin Lai, Yi Geng
Frontiers in Microbiology.2024;[Epub] CrossRef
Rcs signal transduction system in Escherichia coli: Composition, related functions, regulatory mechanism, and applications
Zeyu Li, Yingying Zhu, Wenli Zhang, Wanmeng Mu
Microbiological Research.2024; 285: 127783. CrossRef
Identification of genes used by Escherichia coli to mitigate climatic stress conditions
Styliani Roufou, Sholeem Griffin, Lydia Katsini, Monika Polańska, Jan F.M. Van Impe, Panagiotis Alexiou, Vasilis P. Valdramidis
Gene Reports.2024; 36: 101998. CrossRef
The Role of Propionate-Induced Rearrangement of Membrane Proteins in the Formation of the Virulent Phenotype of Crohn’s Disease-Associated Adherent-Invasive Escherichia coli
Olga V. Pobeguts, Maria A. Galyamina, Elena V. Mikhalchik, Sergey I. Kovalchuk, Igor P. Smirnov, Alena V. Lee, Lyubov Yu. Filatova, Kirill V. Sikamov, Oleg M. Panasenko, Alexey Yu. Gorbachev
International Journal of Molecular Sciences.2024; 25(18): 10118. CrossRef
CpxAR two-component system contributes to virulence properties of Cronobacter sakazakii
Tong Jin, Xiangjun Zhan, Liuxin Pang, Bo Peng, Xinpeng Zhang, Wenxiu Zhu, Baowei Yang, Xiaodong Xia
Food Microbiology.2024; 117: 104393. CrossRef
Breaking Barriers: Exploiting Envelope Biogenesis and Stress Responses to Develop Novel Antimicrobial Strategies in Gram-Negative Bacteria
Renu Bisht, Pierre D. Charlesworth, Paola Sperandeo, Alessandra Polissi
Pathogens.2024; 13(10): 889. CrossRef
The protective role of potassium in the adaptation of Pseudomonas protegens SN15-2 to hyperosmotic stress
Jian Wang, Yaping Wang, Shouquan Lu, Haibo Lou, XiaoBing Wang, Wei Wang
Microbiological Research.2024; 289: 127887. CrossRef
Bacterial Regulatory Mechanisms for the Control of Cellular Processes: Simple Organisms’ Complex Regulation
Jin-Won Lee
Journal of Microbiology.2023; 61(3): 273. CrossRef
Physiological and Transcriptomic Analyses of Escherichia coli Serotype O157:H7 in Response to Rhamnolipid Treatment
Shuo Yang, Lan Ma, Xiaoqing Xu, Qing Peng, Huiying Zhong, Yuxin Gong, Linbo Shi, Mengxin He, Bo Shi, Yu Qiao
Microorganisms.2023; 11(8): 2112. CrossRef

Membrane Proteins as a Regulator for Antibiotic Persistence in Gram‑Negative Bacteria: Jia Xin Yee , Juhyun Kim , Jinki Yeom; J. Microbiol. 2023;61(3):331-341. Published online February 17, 2023; DOI: https://doi.org/10.1007/s12275-023-00024-w

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Abstract PDF

Antibiotic treatment failure threatens our ability to control bacterial infections that can cause chronic diseases. Persister bacteria are a subpopulation of physiological variants that becomes highly tolerant to antibiotics. Membrane proteins play crucial roles in all living organisms to regulate cellular physiology. Although a diverse membrane component involved in persistence can result in antibiotic treatment failure, the regulations of antibiotic persistence by membrane proteins has not been fully understood. In this review, we summarize the recent advances in our understanding with regards to membrane proteins in Gram-negative bacteria as a regulator for antibiotic persistence, highlighting various physiological mechanisms in bacteria.

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Cardamom essential oil-loaded zinc oxide nanoparticles: A sustainable antimicrobial strategy against multidrug-resistant foodborne pathogens
Mabrouk Sobhy, Tamer Elsamahy, Esraa A. Abdelkarim, Ebtihal Khojah, Haiying Cui, Lin Lin
Microbial Pathogenesis.2025; 205: 107661. CrossRef
Amino Acid and Au(III) Self-Assembled Supramolecular Nanozymes for Antimicrobial Applications
Yunzhu Xu, Dahai Hou, Min Zhao, Tong Zhao, Yong Ma, Yafeng Zhang, Yang Guo, Weiwei Tao, Hui Wang
ACS Applied Nano Materials.2024; 7(19): 22505. CrossRef
PhoPQ-mediated lipopolysaccharide modification governs intrinsic resistance to tetracycline and glycylcycline antibiotics in Escherichia coli
Byoung Jun Choi, Umji Choi, Dae-Beom Ryu, Chang-Ro Lee, Mehrad Hamidian, You-Hee Cho
mSystems.2024;[Epub] CrossRef
Bacterial Regulatory Mechanisms for the Control of Cellular Processes: Simple Organisms’ Complex Regulation
Jin-Won Lee
Journal of Microbiology.2023; 61(3): 273. CrossRef

Journal Articles

Construction of high-density transposon mutant library of Staphylococcus aureus using bacteriophage ϕ11: Wonsik Lee; J. Microbiol. 2022;60(12):1123-1129. Published online November 24, 2022; DOI: https://doi.org/10.1007/s12275-022-2476-2

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Transposon mutant libraries are an important resource to study bacterial metabolism and pathogenesis. The fitness analysis of mutants in the libraries under various growth conditions provides important clues to study the physiology and biogenesis of structural components of a bacterial cell. A transposon library in conjunction with next-generation sequencing techniques, collectively named transposon sequencing (Tnseq), enables high-throughput genome profiling and synthetic lethality analysis. Tn-seq has also been used to identify essential genes and to study the mode of action of antibacterials. To construct a high-density transposon mutant library, an efficient delivery system for transposition in a model bacterium is essential. Here, I describe a detailed protocol for generating a high-density phage-based transposon mutant library in a Staphylococcus aureus strain, and this protocol is readily applicable to other S. aureus strains including USA300 and MW2.

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Optimizing phage-based mutant recovery and minimizing heat effect in the construction of transposon libraries in Staphylococcus aureus
Sally W. Yousief, Nader Abdelmalek, Bianca Paglietti
Scientific Reports.2024;[Epub] CrossRef

Mutational analysis on stable expression and LasB inhibition of LasB propeptide in Pseudomonas aeruginosa: Youngsun Shin , Xi-Hui Li , Cheol Seung Lee , Joon-Hee Lee; J. Microbiol. 2022;60(7):727-734. Published online May 25, 2022; DOI: https://doi.org/10.1007/s12275-022-1671-5

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Three major proteases, elastase B (LasB), protease IV (PIV), and elastase A (LasA) expressed in Pseudomonas aeruginosa play important roles in infections and pathogeneses. These are activated by a proteolytic cascade initiated by the activation of LasB. In this study, we investigated whether LasB could be inhibited using its propeptide (LasBpp). Although LasA and PIV were inhibited by their propeptides, LasB was not inhibited by purified LasBpp because LasB degraded LasBpp. To address this problem, mutant LasBpp variants were constructed to obtain a mutant LasBpp resistant to LasB degradation. A C-terminal deletion series of LasBpp was tested in vivo, and two positive candidates, T2 and T2-1, were selected. However, both caused growth retardation and were unstably expressed in vivo. Since deleting the C-terminal end of LasBpp significantly affected its stable expression, substitution mutations were introduced at the two amino acids near the truncation site of T2-1. The resulting mutants, LasBppE172D, LasBppG173A, and LasBppE172DG173A, significantly diminished LasB activity when overexpressed in vivo and were stably expressed in MW1, a quorum sensing mutant that does not produce LasB. In vitro analysis showed that purified LasBppE172DG173A inhibited LasB activity to a small extent. Summarizing, Cterminal modification of LasBpp profoundly affected the stable expression of LasBpp, and little enhanced the ability of LasBpp to resist degradation by LasB.

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LasB activation in Pseudomonas aeruginosa: Quorum sensing-mediated release of an auto-activation inhibitor
Cheol Seung Lee, Xi-Hui Li, Chae-Ran Jeon, Joon-Hee Lee
Journal of Microbiology.2025; 63(2): e2411005. CrossRef

Meta-Analysis

Proposal of a health gut microbiome index based on a meta-analysis of Korean and global population datasets: Hyun-Seok Oh , Uigi Min , Hyejin Jang , Namil Kim , Jeongmin Lim , Mauricio Chalita , Jongsik Chun; J. Microbiol. 2022;60(5):533-549. Published online March 31, 2022; DOI: https://doi.org/10.1007/s12275-022-1526-0

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The disruption of the human gut microbiota has been linked to host health conditions, including various diseases. However, no reliable index for measuring and predicting a healthy microbiome is currently available. Here, the sequencing data of 1,663 Koreans were obtained from three independent studies. Furthermore, we pooled 3,490 samples from public databases and analyzed a total of 5,153 fecal samples. First, we analyzed Korean gut microbiome covariates to determine the influence of lifestyle on variation in the gut microbiota. Next, patterns of microbiota variations across geographical locations and disease statuses were confirmed using a global cohort and disease data. Based on comprehensive comparative analysis, we were able to define three enterotypes among Korean cohorts, namely, Prevotella type, Bacteroides type, and outlier type. By a thorough categorization of dysbiosis and the evaluation of microbial characteristics using multiple datasets, we identified a wide spectrum of accuracy levels in classifying health and disease states. Using the observed microbiome patterns, we devised an index named the gut microbiome index (GMI) that could consistently predict health conditions from human gut microbiome data. Compared to ecological metrics, the microbial marker index, and machine learning approaches, GMI distinguished between healthy and non-healthy individuals with a higher accuracy across various datasets. Thus, this study proposes a potential index to measure health status of gut microbiome that is verified from multiethnic data of various diseases, and we expect this model to facilitate further clinical application of gut microbiota data in future.

Citations

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Methodology for Assessing Patient Centricity and Data Integrity in Clinical Trials With Decentralized Elements: A Pilot Trial on Mastic Gum
Jiyeon Park, Ki Young Huh, Jae‐Yong Chung, Kyung‐Sang Yu
Clinical and Translational Science.2025;[Epub] CrossRef
A comparison of the prevalence of respiratory pathogens and opportunistic respiratory pathogenic profile of ‘clean’ and ‘unclean’ removable dental prostheses
Tong Wah Lim, Shi Huang, Yufeng Zhang, Michael Francis Burrow, Colman McGrath
Journal of Dentistry.2024; 145: 104968. CrossRef
Characterization of pathogenic microbiome on removable prostheses with different levels of cleanliness using 2bRAD-M metagenomic sequencing
Tong Wah Lim, Shi Huang, Yuesong Jiang, Yufeng Zhang, Michael Francis Burrow, Colman McGrath
Journal of Oral Microbiology.2024;[Epub] CrossRef
Gut microbial signatures in clinically stable ulcerative colitis according to the mucosal state and associated symptoms
Soyoung Kim, Yeonjae Jung, Seung Bum Lee, Hyun‐Seok Oh, Sung Noh Hong
Journal of Gastroenterology and Hepatology.2024; 39(2): 319. CrossRef
Difference in gut microbial dysbiotic patterns between body-first and brain-first Parkinson's disease
Don Gueu Park, Woorim Kang, In-Ja Shin, Mauricio Chalita, Hyun-Seok Oh, Dong-Wook Hyun, Hyun Kim, Jongsik Chun, Young-Sil An, Eun Jeong Lee, Jung Han Yoon
Neurobiology of Disease.2024; 201: 106655. CrossRef
Should Routine Diagnostics Implement Gut Microbiota Analysis?
Giuseppe Guido Maria Scarlata, Ludovico Abenavoli
The International Journal of Gastroenterology and Hepatology Diseases.2024;[Epub] CrossRef
Feasibility study for a fully decentralized clinical trial in participants with functional constipation symptoms
Ki Young Huh, Woo Kyung Chung, Jiyeon Park, SeungHwan Lee, Min‐Gul Kim, Jaeseong Oh, Kyung‐Sang Yu
Clinical and Translational Science.2023; 16(11): 2177. CrossRef
Predicting Personalized Responses to Dietary Fiber Interventions: Opportunities for Modulation of the Gut Microbiome to Improve Health
Car Reen Kok, Devin Rose, Robert Hutkins
Annual Review of Food Science and Technology.2023; 14(1): 157. CrossRef
Effects of the multidomain intervention with nutritional supplements on cognition and gut microbiome in early symptomatic Alzheimer’s disease: a randomized controlled trial
Eun Hye Lee, Geon Ha Kim, Hee Kyung Park, Hae Jin Kang, Yoo Kyoung Park, Hye Ah Lee, Chang Hyung Hong, So Young Moon, Woorim Kang, Hyun-Seok Oh, Hai-Jeon Yoon, Seong Hye Choi, Jee Hyang Jeong
Frontiers in Aging Neuroscience.2023;[Epub] CrossRef
Fecal microbial signatures of healthy Han individuals from three bio-geographical zones in Guangdong
Litao Huang, Liting Deng, Changhui Liu, Enping Huang, Xiaolong Han, Cheng Xiao, Xiaomin Liang, Huilin Sun, Chao Liu, Ling Chen
Frontiers in Microbiology.2022;[Epub] CrossRef

Journal Articles

Lactobacillus plantarum-derived metabolites sensitize the tumorsuppressive effects of butyrate by regulating the functional expression of SMCT1 in 5-FU-resistant colorectal cancer cells: Hye-Ju Kim , JaeJin An , Eun-Mi Ha; J. Microbiol. 2022;60(1):100-117. Published online December 29, 2021; DOI: https://doi.org/10.1007/s12275-022-1533-1

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A critical obstacle to the successful treatment of colorectal cancer (CRC) is chemoresistance. Chemoresistant CRC cells contribute to treatment failure by providing a mechanism of drug lethargy and modifying chemoresistance-associated molecules. The gut microbiota provide prophylactic and therapeutic effects by targeting CRC through anticancer mechanisms. Among them, Lactobacillus plantarum contributes to the health of the host and is clinically effective in treating CRC. This study confirmed that 5-fluorouracil (5-FU)-resistant CRC HCT116 (HCT116/5FUR) cells acquired butyrateinsensitive properties. To date, the relationship between 5- FU-resistant CRC and butyrate resistance has not been elucidated. Here, we demonstrated that the acquisition of butyrate resistance in HCT116/5FUR cells was strongly correlated with the inhibition of the expression and function of SMCT1, a major transporter of butyrate in colonocytes. L. plantarum-cultured cell-free supernatant (LP) restored the functional expression of SMCT1 in HCT116/5FUR cells, leading to butyrate-induced antiproliferative effect and apoptosis. These results suggest that LP has a synergistic effect on the SMCT1/butyrate-mediated tumor suppressor function and is a potential chemosensitizer to overcome dual 5-FU and butyrate resistance in HCT116 cells.

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Advancements in the Pathogenesis, Diagnosis, and Therapeutic Implications of Intestinal Bacteria
Duofei Lu, Xianxiong Ma, Kaixiong Tao, Hongwei Lei
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Short-chain fatty acids inhibit the biofilm formation of Streptococcus gordonii through negative regulation of competence-stimulating peptide signaling pathway: Taehwan Park , Jintaek Im , A Reum Kim , Dongwook Lee , Sungho Jeong , Cheol-Heui Yun , Seung Hyun Han; J. Microbiol. 2021;59(12):1142-1149. Published online December 4, 2021; DOI: https://doi.org/10.1007/s12275-021-1576-8

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Streptococcus gordonii, a Gram-positive commensal bacterium, is an opportunistic pathogen closely related to initiation and progression of various oral diseases, such as periodontitis and dental caries. Its biofilm formation is linked with the development of such diseases by enhanced resistance against antimicrobial treatment or host immunity. In the present study, we investigated the effect of short-chain fatty acids (SCFAs) on the biofilm formation of S. gordonii. SCFAs, including sodium acetate (NaA), sodium propionate (NaP), and sodium butyrate (NaB), showed an effective inhibitory activity on the biofilm formation of S. gordonii without reduction in bacterial growth. SCFAs suppressed S. gordonii biofilm formation at early time points whereas SCFAs did not affect its preformed biofilm. A quorum-sensing system mediated by competence-stimulating peptide (CSP) is known to regulate biofilm formation of streptococci. Interestingly, SCFAs substantially decreased mRNA expression of comD and comE, which are CSP-sensor and its response regulator responsible for CSP pathway, respectively. Although S. gordonii biofilm formation was enhanced by exogenous synthetic CSP treatment, such effect was not observed in the presence of SCFAs. Collectively, these results suggest that SCFAs have an anti-biofilm activity on S. gordonii through inhibiting comD and comE expression which results in negative regulation of CSP quorum-sensing system. SCFAs could be an effective anti-biofilm agent against S. gordonii for the prevention of oral diseases.

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[PROTOCOL] Flow cytometric monitoring of the bacterial phenotypic diversity in aquatic ecosystems: Jin-Kyung Hong , Soo Bin Kim , Seok Hyun Ahn , Yongjoo Choi , Tae Kwon Lee; J. Microbiol. 2021;59(10):879-885. Published online September 23, 2021; DOI: https://doi.org/10.1007/s12275-021-1443-7

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Flow cytometry is a promising tool used to identify the phenotypic features of bacterial communities in aquatic ecosystems by measuring the physical and chemical properties of cells based on their light scattering behavior and fluorescence. Compared to molecular or culture-based approaches, flow cytometry is suitable for the online monitoring of microbial water quality because of its relatively simple sample preparation process, rapid analysis time, and high-resolution phenotypic data. Advanced statistical techniques (e.g., denoising and binning) can be utilized to successfully calculate phenotypic diversity by processing the scatter data obtained from flow cytometry. These phenotypic diversities were well correlated with taxonomic-based diversity computed using nextgeneration 16S RNA gene sequencing. The protocol provided in this paper should be a useful guide for a fast and reliable flow cytometric monitoring of bacterial phenotypic diversity in aquatic ecosystems.

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Enhancing Bacterial Phenotype Classification Through the Integration of Autogating and Automated Machine Learning in Flow Cytometric Analysis
In Jae Jeong, Jin‐Kyung Hong, Young Jun Bae, Tea Kwon Lee
Cytometry Part A.2025; 107(3): 203. CrossRef
Assessing long-term ecological impacts of PCE contamination in groundwater using a flow cytometric fingerprint approach
Jin-Kyung Hong, Soo Bin Kim, Gui Nam Wee, Bo Ram Kang, Jee Hyun No, Susmita Das Nishu, Joonhong Park, Tae Kwon Lee
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Gui Nam Wee, Eun Sun Lyou, Susmita Das Nishu, Tae Kwon Lee
Frontiers in Microbiology.2023;[Epub] CrossRef

Full-repertoire comparison of the microscopic objects composing the human gut microbiome with sequenced and cultured communities: Edmond Kuete Yimagou , Jean-Pierre Baudoin , Rita Abou Abdallah , Fabrizio Di Pinto , Jacques Yaacoub Bou Khalil , Didier Raoult; J. Microbiol. 2020;58(5):377-386. Published online April 11, 2020; DOI: https://doi.org/10.1007/s12275-020-9365-3

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The study of the human gut microbiome is essential in microbiology and infectious diseases as specific alterations in the gut microbiome might be associated with various pathologies, such as chronic inflammatory disease, intestinal infection and colorectal cancer. To identify such dysregulations, several strategies are being used to create a repertoire of the microorganisms composing the human gut microbiome. In this study, we used the “microscomics” approach, which consists of creating an ultrastructural repertoire of all the cell-like objects composing stool samples from healthy donors using transmission electron microscopy (TEM). We used TEM to screen ultrathin sections of 8 resin-embedded stool samples. After exploring hundreds of micrographs, we managed to elaborate ultrastructural categories based on morphological criteria or features. This approach explained many inconsistencies observed with other techniques, such as metagenomics and culturomics. We highlighted the value of our cultureindependent approach by comparing our microscopic images to those of cultured bacteria and those reported in the literature. This study helped to detect “minimicrobes” Candidate Phyla Radiation (CPR) for the first time in human stool samples. This “microscomics” approach is non-exhaustive but complements already existing approaches and adds important data to the puzzle of the microbiota.

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Tips and tricks for gut microbiota investigation using scanning electron microscopy (SEM): going from sample preparation to imaging and landscape analysis
Meriem Boukili, Omar Zmerli, Florence Fenollar, Sara Bellali, Jacques Bou Khalil
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Candidate Phyla Radiation, an Underappreciated Division of the Human Microbiome, and Its Impact on Health and Disease
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Comparative genomic analysis of selenium utilization traits in different marine environments: Muhammad Farukh; J. Microbiol. 2020;58(2):113-122. Published online January 29, 2020; DOI: https://doi.org/10.1007/s12275-020-9250-0

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Selenium (Se) is an essential trace element for many organisms, which is required in the biosynthesis of proteins with selenocysteine, tRNAs with selenouridine, and certain enzymes with Se as a cofactor. Recent large-scale metagenomics projects provide a unique opportunity for studying the global trends of Se utilization in marine environments. Here, we analyzed samples from different marine microbial communities, revealed by the Tara Oceans project, to characterize the Se utilization traits. We found that the selenophosphate synthetase gene, which defines the overall Se utilization, and Se utilization traits are present in all samples. Regions with samples rich and poor in Se utilization traits were categorized. From the analysis of environmental factors, the mesopelagic zone and high temperature (> 15°C) of water are favorable, while geographical location has little influence on Se utilization. All Se utilization traits showed a relatively independent occurrence. The taxonomic classification of Se traits shows that most of the sequences corresponding to Se utilization traits belong to the phylum Proteobacteria. Overall, our study provides useful insights into the general features of Se utilization in ocean samples and may help to understand the evolutionary dynamics of Se utilization in different marine environments.

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The selenophosphate synthetase family: A review
Bruno Manta, Nadezhda E Makarova, Marco Mariotti
Free Radical Biology and Medicine.2022; 192: 63. CrossRef
Selenium Metabolism and Selenoproteins in Prokaryotes: A Bioinformatics Perspective
Yan Zhang, Jiao Jin, Biyan Huang, Huimin Ying, Jie He, Liang Jiang
Biomolecules.2022; 12(7): 917. CrossRef
Uses of Selenium Nanoparticles in the Plant Production
Iqra Bano, Sylvie Skalickova, Hira Sajjad, Jiri Skladanka, Pavel Horky
Agronomy.2021; 11(11): 2229. CrossRef

Development of a neutralization assay based on the pseudotyped chikungunya virus of a Korean isolate: Woo-Chang Chung , Kwang Yeon Hwang , Suk-Jo Kang , Jae-Ouk Kim , Moon Jung Song; J. Microbiol. 2020;58(1):46-53. Published online November 25, 2019; DOI: https://doi.org/10.1007/s12275-020-9384-0

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The Chikungunya virus (CHIKV) belongs to the Alphavirus genus of Togaviridae family and contains a positive-sense single stranded RNA genome. Infection by this virus mainly causes sudden high fever, rashes, headache, and severe joint pain that can last for several months or years. CHIKV, a mosquito- borne arbovirus, is considered a re-emerging pathogen that has become one of the most pressing global health concerns due to a rapid increase in epidemics. Because handling of CHIKV is restricted to Biosafety Level 3 (BSL-3) facilities, the evaluation of prophylactic vaccines or antivirals has been substantially hampered. In this study, we first identified the whole structural polyprotein sequence of a CHIKV strain isolated in South Korea (KNIH/2009/77). Phylogenetic analysis showed that this sequence clustered within the East/ Central/South African CHIKV genotype. Using this sequence information, we constructed a CHIKV-pseudotyped lentivirus expressing the structural polyprotein of the Korean CHIKV isolate (CHIKVpseudo) and dual reporter genes of green fluorescence protein and luciferase. We then developed a pseudovirus-based neutralization assay (PBNA) using CHIKVpseudo. Results from this assay compared to those from the conventional plaque reduction neutralization test showed that our PBNA was a reliable and rapid method to evaluate the efficacy of neutralizing antibodies. More importantly, the neutralizing activities of human sera from CHIKVinfected individuals were quantitated by PBNA using CHIKVpseudo. Taken together, these results suggest that our PBNA for CHIKV may serve as a useful and safe method for testing the neutralizing activity of antibodies against CHIKV in BSL-2 facilities.

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Sofiia N. Rizatdinova, Alina E. Ershova, Irina V. Astrakhantseva
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Yao Yan, Fengyuan Zhang, Meng Zou, Hongyu Chen, Jingwen Xu, Shuaiyao Lu, Hongqi Liu
Acta Biochimica et Biophysica Sinica.2024; 56(10): 1425. CrossRef
Facile quantitative diagnostic testing for neutralizing antibodies against Chikungunya virus
Hui-Chung Lin, Shu-Fen Chang, Chien-Ling Su, Huai-Chin Hu, Der-Jiang Chiao, Yu-Lin Hsu, Hsuan-ying Lu, Chang-Chi Lin, Pei-Yun Shu, Szu-Cheng Kuo
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Hui-Chung Lin, Der-Jiang Chiao, Pei-Yun Shu, Hui-Tsu Lin, Chia-Chu Hsiung, Chang-Chi Lin, Szu-Cheng Kuo, Juan E. Ludert
Microbiology Spectrum.2023;[Epub] CrossRef
Preparation and application of chikungunya pseudovirus containing double reporter genes
Chunyan Su, Kaiyun Ding, Jingwen Xu, Jianchao Wu, Jiansheng Liu, Jiayuan Shen, Hongning Zhou, Hongqi Liu
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Hahyoungchilella caricis gen. nov., sp. nov., isolated from a rhizosphere mudflat of a halophyte (Carex scabrifolia), transfer of Thioclava arenosa Thongphrom et al. 2017 to Pseudothioclava as Pseudothioclava arenosa gen. nov., comb. nov. and proposal of Thioclava electrotropha Chang et al. 2018: Young-Ju Kim , Soon Dong Lee; J. Microbiol. 2019;57(12):1048-1055. Published online September 25, 2019; DOI: https://doi.org/10.1007/s12275-019-9260-y

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A Gram-stain-negative, strictly aerobic, marine bacterium, designated GH2-2T, was isolated from a rhizosphere mudflat of a halophyte (Carex scabrifolia) in Gangwha Island, the Republic of Korea. The cells of the organism were oxidase- positive, catalase-positive, flagellated, short rods that grew at 10–40°C, pH 4–10, and 0–13% (w/v) NaCl. The predominant ubiquinone was Q-10. The major polar lipids were phosphatidylcholine, phosphatidylethanolamine, and phosphatidylglycerol. The major fatty acid is C18:1. Phylogenetic analysis based on 16S rRNA gene sequences revealed that the novel isolate formed an independent lineage at the base of the radiation encompassing members of the genus Thioclava, except for Thioclava arenosa. The closest relatives were T. nitratireducens (96.03% sequence similarity) and T. dalianensis (95.97%). The genome size and DNA G+C content were 3.77 Mbp and 59.6 mol%, respectively. Phylogenomic analysis supported phylogenetic distinctness based on 16S rRNA gene sequences. Average nucleotide identity values were 73.6–74.0% between the novel strain and members of the genus Thioclava. On the basis of data obtained from a polyphasic approach, the strain GH2-2T (= KCTC 62124T = DSM 105743T) represents a novel species of a new genus for which the name Hahyoungchilella caricis gen. nov., sp. nov. is proposed. Moreover, the transfer of Thioclava arenosa Thongphrom et al. 2017 to Pseudothioclava gen. nov. as Pseudothioclava arenosa comb. nov. is also proposed. Finally, Thioclava electrotropha Chang et al. 2018 is proposed to be a later heterosynonym of Thioclava sediminum Liu et al. 2017.

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Staphylococcus aureus is the major pathogen leading to bovine mastitis globally while livestock-associated methicillin resistant S. aureus (LA-MRSA) has become a potential threat to public health. MRSA from bovine mastitis is not common but a methicillin susceptible S. aureus (MSSA) genotype, rpoB sequence type (RST)10-2 (RST10-2), is prevalent in Korea. To date, many genomic sequences from S. aureus have been elucidated, but the complete genome sequences of RST10-2 MSSA from bovine mastitis has never been reported. In this study, we determined the complete genome sequence of two RST10-2 MSSA that differ from each other in staphylococcal protein A and molecular prophage types [PMB64-1 (t2489/ mPPT0) and PMB81-4 (t127/mPPT1-2-3)] and conducted a comparative genomics study. The genomic sequences of PMB64-1 and PMB81-4 were more homologous to the representative human RST10-2 strains (MSSA476, MW2 etc.) compared to other RSTs. Most of them shared five common pseudogenes, along with high amino acid identity of four variable virulence genes that were identified in this study. However, PMB64-1 and PMB81-4 acquired different strainspecific pseudogenes and mobile genetic elements than the human strains. The unique pseudogene profile and high identity of the virulence genes were verified in RST10-2 field strains from bovine mastitis. Thus, bovine mastitic RST10-2 MSSA may have an evolutionary relationship with the human RST10- 2 community-associated (CA) MSSA and CA-MRSA strains but may have adapted to cows.

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Differential expression of the major catalase, KatA in the two wild type Pseudomonas aeruginosa strains, PAO1 and PA14: Bi-o Kim , In-Young Chung , You-Hee Cho; J. Microbiol. 2019;57(8):704-710. Published online June 11, 2019; DOI: https://doi.org/10.1007/s12275-019-9225-1

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KatA is the major catalase required for hydrogen peroxide (H2O2) resistance and acute virulence in Pseudomonas aeruginosa PA14, whose transcription is governed by its dual promoters (katAp1 and katAp2). Here, we observed that KatA was not required for acute virulence in another wild type P. aeruginosa strain, PAO1, but that PAO1 exhibited higher KatA expression than PA14 did. This was in a good agreement with the observation that PAO1 was more resistant than PA14 to H2O2 as well as to the antibiotic peptide, polymyxin B (PMB), supposed to involve reactive oxygen species (ROS) for its antibacterial activity. The higher KatA expression in PAO1 than in PA14 was attributed to both katAp1 and katAp2 transcripts, as assessed by S1 nuclease mapping. In addition, it was confirmed that the PMB resistance is attributed to both katAp1 and katAp2 in a complementary manner in PA14 and PAO1, by exploiting the promoter mutants for each -10 box (p1m, p2m, and p1p2m). These results provide an evidence that the two widely used P. aeruginosa strains display different virulence mechanisms associated with OxyR and Anr, which need to be further characterized for better understanding of the critical virulence pathways that may differ in various P. aeruginosa strains.

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Low-density lipoprotein as an opsonin promoting the phagocytosis of Pseudomonas aeruginosa by U937 cells: Yuxin Li , Zhi Liu , Jinli Yang , Ling Liu , Runlin Han; J. Microbiol. 2019;57(8):711-716. Published online May 11, 2019; DOI: https://doi.org/10.1007/s12275-019-8413-3

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Low-density lipoprotein (LDL) was recently reported to be an opsonin, enhancing the phagocytosis of group A Streptococcus (GAS) by human monocytic leukemia U937 cells due to the binding of LDL to some GAS strains. We postulated that LDL might also promote the opsonophagocytosis of Pseudomonas aeruginosa by U937 cells since this bacterium interacts with LDL. In this study, P. aeruginosa (CMCC10104), U937 cells, and human LDL were used in phagocytosis assays to test our hypothesis. Escherichia coli strain BL21, which does not interact with LDL, was used as a negative control. Colony counting and fluorescence microscopy were used to determine the bacterial quantity in the opsonophagocytosis assays. After incubation of U937 cells and P. aeruginosa with LDL (100 μg/ml) for 15 and 30 min, phagocytosis was observed to be increased by 22.71% and 32.90%, respectively, compared to that seen in the LDL-free group. However, LDL did not increase the phagocytosis of E. coli by U937 cells. In addition, we identified CD36 as a major opsonin receptor on U937 cells, since an anti-CD36 monoclonal antibody, but not an anti- CD4 monoclonal antibody, almost completely abolished the opsonophagocytosis of P. aeruginosa by U937 cells.

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Pseudomonas aeruginosa, an opportunistic human pathogen, causes many biofilm-mediated chronic infections. In this study, biofilm structures of various clinical strains of P. aeruginosa isolated from hospitalized patients were examined and their influence on the biofilm-dispersing effects of chemicals was investigated. The clinical isolates formed structurally distinct biofilms that could be classified into three different groups: 1) mushroom-like, 2) thin flat, and 3) thick flat structures. A dispersion of these differently structured biofilms was induced using two biofilm-dispersing agents, anthranilate and sodium nitroprusside (SNP). Although both SNP and anthranilate could disperse all types of biofilms, the thick flat biofilms were dispersed less efficiently than the biofilms of other structures. This suggests that biofilm-dispersing agents have higher potency on the biofilms of porous structures than on densely packed biofilms.

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Circular pellicles formed by Pseudomonas alkylphenolica KL28 are a sophisticated architecture principally designed by matrix substance: Myeong Mi Song , Yaligara Veeranagouda , Munkhtsatsral Ganzorig , Kyoung Lee; J. Microbiol. 2018;56(11):790-797. Published online October 24, 2018; DOI: https://doi.org/10.1007/s12275-018-8252-7

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The colonization of liquid surfaces as floating biofilms or pellicles is a bacterial adaptation to optimally occupy the airliquid (A-L) niche. In aerobic heterotrophs, pellicle formation is beneficial for the utilization of O2 and nonpolar organic compounds. Pseudomonas alkylphenolica KL28, an alkylphenol degrader, forms flat circular pellicles that are 0.3– 0.5 mm in diameter. In this study, we first monitored the pellicle developmental patterns of multicellular organization from the initial settlement stage. The pellicles developed by clonal growth and mutants for flagella and pilus formation established normal pellicles. In contrast, the mutants of an epm gene cluster for biosynthesis of alginate-like polymer were incompetent in cell alignment for initial two-dimensional (2D) pellicle growth, suggesting the role of the Epm polymer as a structural scaffold for pellicle biofilms. Microscopic observation revealed that the initial 2D growth transited to multilayers by an accumulated self-produced extracellular polymeric substance that may exert a constraint force. Electron microscopy and confocal laser scanning microscopy revealed that the fully matured pellicle structures were densly packed with matrix-encased cells displaying distinct arrangements. The cells on the surface of the pellicle were relatively flat, and those inside were longitudinally cross-packed. The extracellular polysaccharide stained by Congo red was denser on the pellicle rim and a thin film was observed in the open spaces, indicative of its role in pellicle flotation. Our results demonstrate that P. alkylphenolica KL28 coordinately dictates the cell arrangements of pellicle biofilms by the controlled growth of constituent cells that accumulate extracellular polymeric substances.

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Review

REVIEW] Antibiotic-resistant clones in Gram-negative pathogens: presence of global clones in Korea: Kwan Soo Ko; J. Microbiol. 2019;57(3):195-202. Published online October 2, 2018; DOI: https://doi.org/10.1007/s12275-019-8491-2

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Antibiotic resistance is a global concern in public health. Antibiotic-resistant clones can spread nationally, internationally, and globally. This review considers representative antibiotic-resistant Gram-negative bacterial clones–CTX-M- 15-producing ST131 in Escherichia coli, extended-spectrum β-lactamase-producing ST11 and KPC-producing ST258 in Klebsiella pneumoniae, IMP-6-producing, carbapenem-resistant ST235 in Pseudomonas aeruginosa, and OXA-23- producing global clone 2 in Acinetobacter baumannii–that have disseminated worldwide, including in Korea. The findings highlight the urgency for systematic monitoring and international cooperation to suppress the emergence and propagation of antibiotic resistance.

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Journal Article

[PROTOCOL] Drosophila melanogaster as a polymicrobial infection model for Pseudomonas aeruginosa and Staphylococcus aureus: Young-Joon Lee , Hye-Jeong Jang , In-Young Chung , You-Hee Cho; J. Microbiol. 2018;56(8):534-541. Published online July 25, 2018; DOI: https://doi.org/10.1007/s12275-018-8331-9

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Non-mammalian infection models have been developed over the last two decades, which is a historic milestone to understand the molecular basis of bacterial pathogenesis. They also provide small-scale research platforms for identification of virulence factors, screening for antibacterial hits, and evaluation of antibacterial efficacy. The fruit fly, Drosophila melanogaster is one of the model hosts for a variety of bacterial pathogens, in that the innate immunity pathways and tissue physiology are highly similar to those in mammals. We here present a relatively simple protocol to assess the key aspects of the polymicrobial interaction in vivo between the human opportunistic pathogens, Pseudomonas aeruginosa and Staphylococcus aureus, which is based on the systemic infection by needle pricking at the dorsal thorax of the flies. After infection, fly survival and bacteremia over time for both P. aeruginosa and S. aureus within the infected flies can be monitored as a measure of polymicrobial virulence potential. The infection takes ~24 h including bacterial cultivation. Fly survival and bacteremia are assessed using the infected flies that are monitored up to ~60 h post-infection. These methods can be used to identify presumable as well as unexpected phenotypes during polymicrobial interaction between P. aeruginosa and S. aureus mutants, regarding bacterial pathogenesis and host immunity.

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Review

[MINIREVIEW] Interdependence between iron acquisition and biofilm formation in Pseudomonas aeruginosa: Donghoon Kang , Natalia V. Kirienko; J. Microbiol. 2018;56(7):449-457. Published online June 14, 2018; DOI: https://doi.org/10.1007/s12275-018-8114-3

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Abstract PDF

Bacterial biofilms remain a persistent threat to human healthcare due to their role in the development of antimicrobial resistance. To combat multi-drug resistant pathogens, it is crucial to enhance our understanding of not only the regulation of biofilm formation, but also its contribution to bacterial virulence. Iron acquisition lies at the crux of these two subjects. In this review, we discuss the role of iron acquisition in biofilm formation and how hosts impede this mechanism to defend against pathogens. We also discuss recent findings that suggest that biofilm formation can also have the reciprocal effect, influencing siderophore production and iron sequestration.

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Journal Articles

Taxonomic description and draft genome of Pseudomonas sediminis sp. nov., isolated from the rhizospheric sediment of Phragmites karka: Pratiksha Behera , Madhusmita Mahapatra , Arman Seuylemezian , Parag Vaishampayan , V. Venkata Ramana , Neetha Joseph , Amaraja Joshi , Yogesh Shouche , Mrutyunjay Suar , Ajit K. Pattnaik , Gurdeep Rastogi; J. Microbiol. 2018;56(7):458-466. Published online June 14, 2018; DOI: https://doi.org/10.1007/s12275-018-7549-x

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The taxonomic position of a Gram-stain-negative, rod-shaped bacterial strain, designated PI11T , isolated from the rhizospheric sediment of Phragmites karka was characterized using a polyphasic approach. Strain PI11T could grow optimally at 1.0% NaCl concentration with pH 7.0 at 30°C and was positive for oxidase and catalase but negative for hydrolysis of starch, casein, and esculin ferric citrate. Phylogenetic analysis of 16S rRNA gene sequences indicated that the strain PI11T belonged to the genus Pseudomonas sharing the highest sequence similarities with Pseudomonas indoloxydans JCM 14246T (99.72%), followed by, Pseudomonas oleovorans subsp. oleovorans DSM 1045T (99.29%), Pseudomonas toyotomiensis JCM 15604T (99.15%), Pseudomonas chengduensis DSM 26382T (99.08%), Pseudomonas oleovorans subsp. lubricantis DSM 21016T (99.08%), and Pseudomonas alcaliphila JCM 10630T (99.01%). Experimental DNA-DNA relatedness between strain PI11T and P. indoloxydans JCM 14246T was 49.4%. The draft genome of strain PI11T consisted of 4,884,839 bp. Average nucleotide identity between the genome of strain PI11T and other closely related type strains ranged between 77.25–90.74%. The polar lipid pattern comprised of phosphatidylglycerol, diphosphatidylglycerol, and phosphatidylcholine. The major (> 10%) cellular fatty acids were C18:1 ω6c/ω7c, C16:1 ω6c/ω7c, and C16:0. The DNA G + C content of strain PI11T was 62.4 mol%. Based on the results of polyphasic analysis, strain PI11T was delineated from other closely related type strains. It is proposed that strain PI11T represents a novel species of the genus Pseudomonas, for which the name Pseudomonas sediminis sp. nov. is proposed. The type strain is PI11T (= KCTC 42576T = DSMZ 100245T).

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Yeon Bee Kim, Joon Yong Kim, Hye Seon Song, Se Hee Lee, Na-Ri Shin, Jin-Woo Bae, Jinjong Myoung, Ki-Eun Lee, In-Tae Cha, Jin-Kyu Rhee, Seong Woon Roh
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A common evolutionary pathway for maintaining quorum sensing in Pseudomonas aeruginosa: Bai-min Lai , Hui-cong Yan , Mei-zhen Wang , Na Li , Dong-sheng Shen; J. Microbiol. 2018;56(2):83-89. Published online February 2, 2018; DOI: https://doi.org/10.1007/s12275-018-7286-1

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In the bacterium Pseudomonas aeruginosa, the synthesis and secretion of extracellular protease is a typical cooperative behavior regulated by quorum sensing. However, this type of cooperative behavior is easily exploited by other individuals who do not synthesize public goods, which is known as the “tragedy of the commons”. Here P. aeruginosa was inoculated into casein media with different nitrogen salts added. In casein broth, protease (a type of public good) is necessary for bacterial growth. After 30 days of sequential transfer, some groups propagated stably and avoided “tragedy of the commons”. The evolved cooperators who continued to synthesize protease were isolated from these stable groups. By comparing the characteristics of quorum sensing in these cooperators, an identical evolutionary pattern was found. A variety of cooperative behaviors regulated by quorum sensing, such as the synthesis and secretion of protease and signals, were significantly reduced during the process of evolution. Such reductions improved the efficiency of cooperation, helping to prevent cheating. In addition, the production of pyocyanin, which is regulated by the RhlIR system, increased during the process of evolution, possibly due to its role in stabilizing the cooperation. This study contributes towards our understanding of the evolution of quorum sensing of P. aeruginosa.

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To cheat or not to cheat: cheatable and non-cheatable virulence factors in Pseudomonas aeruginosa
Katya Dafne Guadarrama-Orozco, Caleb Perez-Gonzalez, Kokila Kota, Miguel Cocotl-Yañez, Jesús Guillermo Jiménez-Cortés, Miguel Díaz-Guerrero, Mariel Hernández-Garnica, Julia Munson, Frederic Cadet, Luis Esaú López-Jácome, Ángel Yahir Estrada-Velasco, Ana M
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Exoprotease exploitation and social cheating in a Pseudomonas aeruginosa environmental lysogenic strain with a noncanonical quorum sensing system
Daniel Huelgas-Méndez, Daniel Cazares, Luis David Alcaraz, Corina Diana Ceapã, Miguel Cocotl-Yañez, Toya Shotaro, Toshinari Maeda, Ana María Fernández-Presas, Oswaldo Tostado-Islas, Ana Lorena González-Vadillo, Aldo Limones-Martínez, Carlos Eduardo Hernan
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3-Phenylpropan-1-Amine Enhanced Susceptibility of Serratia marcescens to Ofloxacin by Occluding Quorum Sensing
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Tobramycin Adaptation Enhances Policing of Social Cheaters in Pseudomonas aeruginosa
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Seeding Public Goods Is Essential for Maintaining Cooperation in Pseudomonas aeruginosa
Daniel Loarca, Dánae Díaz, Héctor Quezada, Ana Laura Guzmán-Ortiz, Abril Rebollar-Ruiz, Ana María Fernández Presas, Jimena Ramírez-Peris, Rafael Franco-Cendejas, Toshinari Maeda, Thomas K. Wood, Rodolfo García-Contreras
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Pyocyanin Restricts Social Cheating in Pseudomonas aeruginosa
Paulina Castañeda-Tamez, Jimena Ramírez-Peris, Judith Pérez-Velázquez, Christina Kuttler, Ammar Jalalimanesh, Miguel Á. Saucedo-Mora, J. Guillermo Jiménez-Cortés, Toshinari Maeda, Yael González, María Tomás, Thomas K. Wood, Rodolfo García-Contreras
Frontiers in Microbiology.2018;[Epub] CrossRef

Characterization of siderophore produced by Pseudomonas syringae BAF.1 and its inhibitory effects on spore germination and mycelium morphology of Fusarium oxysporum: Sumei Yu , Chunying Teng , Jinsong Liang , Tao Song , Liying Dong , Xin Bai , Yu Jin , Juanjuan Qu; J. Microbiol. 2017;55(11):877-884. Published online October 27, 2017; DOI: https://doi.org/10.1007/s12275-017-7191-z

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In this study, an antagonistic bacterium against Fusarium oxysporum was identified and designated as Pseudomonas syringae strain BAF.1 on the basis of 16S rDNA sequence analysis and physiological-biochemical characteristics. It produced catechol-species siderophore at a molecular weight of 488.59 Da and a maximum amount of 55.27 μg/ml with glucose as a carbon source and asparagine as a nitrogen source at a C/N ratio of 10:1, 30°C and pH 7. The siderophore exhibited prominent antagonistic activity against Fusarium oxysporum with a maximum inhibition rate of 95.24% and had also suppressive effects on other kinds of 11 phytopathogenic fungi in the absence of FeCl3·6H2O. Spore germination was completely inhibited by 50 μl of the siderophorecontaining solution, and the ultrastructures of mycelia and spores were also considerably suppressed by siderophore treatment as established by electron microscopy observation. These results indicate that the siderophore produced by Pseudomonas syringae BAF.1 could be potentially used for biocontrol of pathogenic Fusarium oxysporum.

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Pseudaminobacter granuli sp. nov., isolated from granules used in a wastewater treatment plant: Young Ki Hahn , Minseok S. Kim , Wan-Taek Im; J. Microbiol. 2017;55(8):607-611. Published online July 28, 2017; DOI: https://doi.org/10.1007/s12275-017-7257-y

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A Gram negative, aerobic, non-motile and rod-shaped bacterial strain designated as Gr-2T was isolated from granules used in a wastewater treatment plant in Korea, and its taxonomic position was investigated using a polyphasic approach. Strain Gr-2T grew at 18–37°C (optimum temperature, 30°C) and a pH of 6.0–8.0 (optimum pH, 7.0) on R2A agar medium. Based on 16S rRNA gene phylogeny, the novel strain showed a new branch within the genus Pseudaminobacter of the family Phyllobacteriaceae, and formed clusters with Pseudaminobacter defluvii THI 051T (98.9%) and Pseudaminobacter salicylatoxidans BN12T (98.7%). The G+C content of the genomic DNA was 63.6%. The predominant respiratory quinone was ubiquinone-10 (Q-10) and the major fatty acids were cyclo-C19:0 ω8c, C18:1 ω7c, and iso-C17:0. The overall polar lipid patterns of Gr-2T were similar to those determined for the other Pseudaminobacter species. DNA-DNA relatedness values between strain Gr-2T and its closest phylogenetically neighbors were below 18%. Strain Gr-2T could be differentiated genotypically and phenotypically from the recognized species of the genus Pseudaminobacter. The isolate therefore represents a novel species, for which the name Pseudaminobacter granuli sp. nov. is proposed with the type strain Gr-2T (=KACC 18877T =LMG 29567T).

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Imipenem-resistant Gram-negative bacterial isolates carried by persons upon medical examination in Korea: So Yeon Kim , Sang Yop Shin , Ji-Young Rhee , Kwan Soo Ko; J. Microbiol. 2017;55(8):612-618. Published online July 18, 2017; DOI: https://doi.org/10.1007/s12275-017-6555-8

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Carbapenem-resistant Gram-negative bacteria (CR-GNB) have emerged and disseminated worldwide, become a great concern worldwide including Korea. The prevalence of fecal carriage of imipenem-resistant Gram-negative bacteria (IRGNB) in persons in Korea was investigated. Stool samples were collected from 300 persons upon medical examination. Samples were screened for IR-GNB by using MacConkey agar with 2 μl/ml imipenem. Species were identified by 16S rRNA gene sequence analysis, and antimicrobial susceptibility was determined by the broth microdilution method. In total, 82 IR-GNB bacterial isolates were obtained from 79 (26.3%) out of 300 healthy persons. Multilocus sequence typing analysis showed very high diversity among IR P. aeruginosa, S. maltophilia, and E. cloacae isolates, and pulsedfield gel electrophoresis revealed five main pulsotypes of IR P. mirabilis. As for the presence of metallo-β-lactamases (MBLs), only one IMP-25-producing S. marcescens isolate was identified. Although only one carbapenemase-producing isolate was identified, the high colonization rates with IRGNB isolates in this study is notable because carriers may be a reservoir for the dissemination of resistant pathogens within the community as well as in health care institutions.

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ZntR positively regulates T6SS4 expression in Yersinia pseudotuberculosis: Tietao Wang , Keqi Chen , Fen Gao , Yiwen Kang , Muhammad Tausif Chaudhry , Zhuo Wang , Yao Wang , Xihui Shen; J. Microbiol. 2017;55(6):448-456. Published online March 10, 2017; DOI: https://doi.org/10.1007/s12275-017-6540-2

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The type VI secretion system (T6SS) is a widespread and versatile protein secretion system found in most Gram- negative bacteria. Studies of T6SS have mainly focused on its role in virulence toward host cells and inter-bacterial inter-actions, but studies have also shown that T6SS4 in Yersinia pseudotuberculosis participates in the acquisition of zinc ions to alleviate the accumulation of hydroxyl radicals induced by multiple stressors. Here, by comparing the gene expression patterns of wild-type and zntR mutant Y. pseudotubercu-losis cells using RNA-seq analysis, T6SS4 and 17 other bio-logical processes were found to be regulated by ZntR. T6SS4 was positively regulated by ZntR in Y. pseudotuberculosis, and further investigation demonstrated that ZntR regulates T6SS4 by directly binding to its promoter region. T6SS4 ex-pression is regulated by zinc via ZntR, which maintains in-tracellular zinc homeostasis and controls the concentration of reactive oxygen species to prevent bacterial death under oxidative stress. This study provides new insights into the regulation of T6SS4 by a zinc-dependent transcriptional regu-lator, and it provides a foundation for further investigation of the mechanism of zinc transport by T6SS.

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Antibacterial compound produced by Pseudomonas aeruginosa strain UICC B-40, an endophytic bacterium isolated from Neesia altissima: Rina Hidayati Pratiwi , Iman Hidayat , Muhammad Hanafi , Wibowo Mangunwardoyo; J. Microbiol. 2017;55(4):289-295. Published online January 26, 2017; DOI: https://doi.org/10.1007/s12275-017-6311-0

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This study’s aim was to determine the identity of antibacte-rial compounds produced by Pseudomonas aeruginosa strain UICC B-40 and describe the antibacterial compounds’ me-chanisms of action for damaging pathogenic bacteria cells. Isolation and identification of the compounds were carried out using thin layer chromatography (TLC), nuclear mag-netic resonance (NMR) spectroscopy and liquid chromato-graphy mass spectrometry (LC-MS) analyses. Antibacterial activity was assayed via minimum inhibitory concentration (MIC) and the antibacterial compound mechanism was ob-served morphologically through scanning electron micros-copy (SEM). This study successfully identified the (2E,5E)- phenyltetradeca-2,5-dienoate antibacterial compound (mole-cular weight 300 g/mol), composed of a phenolic ester, fatty acid and long chain of aliphatic group structures. MIC values for this compound were determined at 62.5 μg/ml against Staphylococcus aureus strain ATCC 25923. The mechanism of the compound involved breaking down the bacterial cell walls through the lysis process. The (2E,5E)-phenyltetradeca- 2,5-dienoate compound exhibited inhibitory activity on the growth of Gram-positive bacteria.

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Identification of essential genes of Pseudomonas aeruginosa for its growth in airway mucus: Mohammed Abd Alrahman , Sang Sun Yoon; J. Microbiol. 2017;55(1):68-74. Published online December 30, 2016; DOI: https://doi.org/10.1007/s12275-017-6515-3

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Pseudomonas aeruginosa has been identified as an important causative agent of airway infection, mainly in cystic fibrosis. This disease is characterized by defective mucociliary clearance induced in part by mucus hyper-production. Mucin is a major component of airway mucus and is heavily O-glycosylated, with a protein backbone. Airway infection is known to be established with bacterial adhesion to mucin. However, the genes involved in mucin degradation or utilization remain elusive. In this study, we sought to provide a genetic basis of P. aeruginosa airway growth by identifying those genes. First, using RNASeq analyses, we compared genome-wide expression profiles of PAO1, a prototype P. aeruginosa laboratory strain, grown in M9-mucin (M9M) and M9-glucose (M9G) media. Additionally, a PAO1 transposon (Tn) insertion mutants library was screened for mutants defective in growth in M9M medium. One mutant with a Tn insertion in the xcpU gene (PA3100) was determined to exhibit faulty growth in M9M medium. This gene contributes to the type II secretion system, suggesting that P. aeruginosa uses this secretion system to produce a number of proteins to break down and assimilate the mucin molecule. Furthermore, we screened the PAO1 genome for genes with protease activity. Of 13 mutants, one with mutation in PA3247 gene exhibited defective growth in M9M, suggesting that the PA3247-encoded protease plays a role in mucin utilization. Further mechanistic dissection of this particular process will reveal new drug targets, the inhibition of which could control recalcitrant P. aeruginosa infections.

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Citrobacter rodentium possesses a functional type II secretion system necessary for successful host infection
Z Krekhno, SE Woodward, A Serapio-Palacios, J Peña-Díaz, KM Moon, LJ Foster, BB Finlay
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Cross-talk between cancer and Pseudomonas aeruginosa mediates tumor suppression
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Pseudomonas aeruginosa type IV pili actively induce mucus contraction to form biofilms in tissue-engineered human airways
Tamara Rossy, Tania Distler, Lucas A. Meirelles, Joern Pezoldt, Jaemin Kim, Lorenzo Talà, Nikolaos Bouklas, Bart Deplancke, Alexandre Persat, Victor Sourjik
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Impact of diet and the bacterial microbiome on the mucous barrier and immune disorders
Charlotte A. Alemao, Kurtis F. Budden, Henry M. Gomez, Saima F. Rehman, Jacqueline E. Marshall, Shakti D. Shukla, Chantal Donovan, Samuel C. Forster, Ian A. Yang, Simon Keely, Elizabeth R. Mann, Emad M. El Omar, Gabrielle T. Belz, Philip M. Hansbro
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Daniel J. Hassett, Rhett A. Kovall, Michael J. Schurr, Nalinikanth Kotagiri, Harshita Kumari, Latha Satish
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Yuki Fukaya, Miho Takemura, Takashi Koyanagi, Takashi Maoka, Kazutoshi Shindo, Norihiko Misawa
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Evolutionary conservation of the antimicrobial function of mucus: a first defence against infection
Cassie R Bakshani, Ana L Morales-Garcia, Mike Althaus, Matthew D Wilcox, Jeffrey P Pearson, John C Bythell, J Grant Burgess
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Identification of D-amino acid dehydrogenase as an upstream regulator of the autoinduction of a putative acyltransferase in Corynebacterium glutamicum: Jung-Hoon Lee , Yong-Jae Kim , Hee-Sung Shin , Heung-Shick Lee , Shouguang Jin , Un-Hwan Ha; J. Microbiol. 2016;54(6):432-439. Published online May 27, 2016; DOI: https://doi.org/10.1007/s12275-016-6046-3

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Expression of a putative acyltransferase encoded by NCgl- 0350 of Corynebacterium glutamicum is induced by cell-free culture fluids obtained from stationary-phase growth of both C. glutamicum and Pseudomonas aeruginosa, providing evidence for interspecies communication. Here, we further confirmed that such communication occurs by showing that acyltransferase expression is induced by culture fluid obtained from diverse Gram-negative and -positive bacterial strains, including Escherichia coli, Salmonella Typhimurium, Bacillus subtilis, Staphylococcus aureus, Mycobacterium sp. strain JC1, and Mycobacterium smegmatis. A homologous acyltransferase encoded by PA5238 of P. aeruginosa was also induced by fluids obtained from P. aeruginosa as well as other bacterial strains, as observed for NCgl0350 of C. glutamicum. Because C. glutamicum is difficult to study using molecular approaches, the homologous gene PA5238 of P. aeruginosa was used to identify PA5309 as an upstream regulator of expression. A homologous D-amino acid dehydrogenase encoded by NCgl- 2909 of C. glutamicum was cloned based on amino acid similarity to PA5309, and its role in the regulation of NCgl0350 expression was confirmed. Moreover, NCgl2909 played positive roles in growth of C. glutamicum. Thus, we identified a D-amino acid dehydrogenase as an upstream regulator of the autoinduction of a putative acyltransferase in C. glutamicum.

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Enhanced Bacterial Growth and Gene Expression of D-Amino Acid Dehydrogenase With D-Glutamate as the Sole Carbon Source
Takeshi Naganuma, Yoshiakira Iinuma, Hitomi Nishiwaki, Ryota Murase, Kazuo Masaki, Ryosuke Nakai
Frontiers in Microbiology.2018;[Epub] CrossRef

Review

MINIREVIEW] Biofilm dispersion in Pseudomonas aeruginosa: Soo-Kyoung Kim , Joon-Hee Lee; J. Microbiol. 2016;54(2):71-85. Published online February 2, 2016; DOI: https://doi.org/10.1007/s12275-016-5528-7

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In recent decades, many researchers have written numerous articles about microbial biofilms. Biofilm is a complex community of microorganisms and an example of bacterial group behavior. Biofilm is usually considered a sessile mode of life derived from the attached growth of microbes to surfaces, and most biofilms are embedded in self-produced extracellular matrix composed of extracellular polymeric substances (EPSs), such as polysaccharides, extracellular DNAs (eDNA), and proteins. Dispersal, a mode of biofilm detachment indicates active mechanisms that cause individual cells to separate from the biofilm and return to planktonic life. Since biofilm cells are cemented and surrounded by EPSs, dispersal is not simple to do and many researchers are now paying more attention to this active detachment process. Unlike other modes of biofilm detachment such as erosion or sloughing, which are generally considered passive processes, dispersal occurs as a result of complex spatial differentiation and molecular events in biofilm cells in response to various environmental cues, and there are many biological reasons that force bacterial cells to disperse from the biofilms. In this review, we mainly focus on the spatial differentiation of biofilm that is a prerequisite for dispersal, as well as environmental cues and molecular events related to the biofilm dispersal. More specifically, we discuss the dispersal-related phenomena and mechanisms observed in Pseudomonas aeruginosa, an important opportunistic human pathogen and representative model organism for biofilm study.

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Research Support, Non-U.S. Gov'ts

The hrp pathogenicity island of Pseudomonas syringae pv. tomato DC3000 is induced by plant phenolic acids: Jun Seung Lee , Hye Ryun Ryu , Ji Young Cha , Hyung Suk Baik; J. Microbiol. 2015;53(10):725-731. Published online October 2, 2015; DOI: https://doi.org/10.1007/s12275-015-5256-4

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Plants produce a wide array of antimicrobial compounds, such as phenolic compounds, to combat microbial pathogens. The hrp PAI is one of the major virulence factors in the plant pathogen, Pseudomonas syringae. A major role of hrp PAI is to disable the plant defense system during bacterial invasion. We examined the influence of phenolic compounds on hrp PAI gene expression at low and high concentrations. There was approximately 2.5 times more hrpA and hrpZ mRNA in PtoDC3000 that was grown in minimal media (MM) supplemented with 10 μM of ortho-coumaric acid than in PtoDC3000 grown in MM alone. On the other hand, a significantly lower amount of hrpA mRNA was observed in bacteria grown in MM supplemented with a high concentration of phenolic compounds. To determine the regulation pathway for hrp PAI gene expression, we performed qRTPCR using gacS, gacA, and hrpS deletion mutants.

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Roles of RpoS in Yersinia pseudotuberculosis stress survival, motility, biofilm formation and type VI secretion system expression: Jingyuan Guan , Xiao Xiao , Shengjuan Xu , Fen Gao , Jianbo Wang , Tietao Wang , Yunhong Song , Junfeng Pan , Xihui Shen , Yao Wang; J. Microbiol. 2015;53(9):633-642. Published online August 27, 2015; DOI: https://doi.org/10.1007/s12275-015-0099-6

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Abstract PDF

RpoS (σS), the stationary phase/stress σ factor, controls the expression of a large number of genes involved in cellular responses to a variety of stresses. However, the role of RpoS appears to differ in different bacteria. While RpoS is an important regulator of flagellum biosynthesis, it is associated with biofilm development in Edwardsiella tarda. Biofilms are dense communities formed by bacteria and are important for microbe survival under unfavorable conditions. The type VI secretion system (T6SS) discovered recently is reportedly associated with several phenotypes, ranging from biofilm formation to stress sensing. For example, Vibrio anguillarum T6SS was proposed to serve as a sensor for extracytoplasmic signals and modulates RpoS expression and stress response. In this study, we investigated the physiological roles of RpoS in Yersinia pseudotuberculosis, including bacterial survival under stress conditions, flagella formation, biofilm development and T6SS expression. We found that RpoS is important in resistance to multiple stressors–including H2O2, acid, osmotic and heat shock–in Y. pseudotuberculosis. In addition, our study showed that RpoS not only modulates the expression of T6SS but also regulates flagellum formation by positively controlling the flagellar master regulatory gene flhDC, and affects the formation of biofilm on Caenorhabditis elegans by regulating the synthesis of exopolysaccharides. Taken together, these results show that RpoS plays a central role in cell fitness under several adverse conditions in Y. pseudotuberculosis.

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Transcription level analysis of intracellular Burkholderia pseudomallei illustrates the role of BPSL1502 during bacterial interaction with human lung epithelial cells: Teerasit Techawiwattanaboon , Tanachaporn Bartpho , Rasana Wongratanacheewin Sermswan , Sorujsiri Chareonsudjai; J. Microbiol. 2015;53(2):134-140. Published online January 28, 2015; DOI: https://doi.org/10.1007/s12275-015-4522-9

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Melioidosis caused by Burkholderia pseudomallei is a globally important disease of increasing concern according to high
case
-fatality rate and epidemic spreading. The ability of B. pseudomallei to attach and invade host cells and subsequently survive intracellularly has stimulated many questions concerning the comprehension of bacterial pathogenesis progression. Transcription levels of intracellular B. pseudomallei genes in human lung epithelial cells were therefore analyzed using bioinformatic tools, RT-PCR and real time RT-PCR. Here, it is reported that the identification of bpsl1502, encoding B. pseudomallei SurE (stationary phase survival protein E) located in a global transcriptional regulation operon was accomplished. The up-regulation of B. pseudomallei SurE was demonstrated during intracellular survival of A549 cells at 12, 18, and 24 h post-infection. To investigate the role of this protein, a B. pseudomallei SurE defective mutant was constructed. The invasion and initial survival of the SurE mutants within the A549 cells were impaired. There was no difference, however, between the growth of B. pseudomallei SurE mutant as compared to the wild type in Luria-Bertani culture. These data suggest that SurE may assist B. pseudomallei host cells invade and facilitate early intracellular infection but is not crucial during the stationary growth phase. The identification of B. pseudomallei SurE provides more information of bacterial strategy during an early step of the pathogenesis process of melioidosis.

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Metabolomics Analysis of Bacterial PathogenBurkholderia thailandensisand Mammalian Host Cells in Co-culture
Nicole Aiosa, Anupama Sinha, Olakunle A. Jaiyesimi, Ricardo R. da Silva, Steven S. Branda, Neha Garg
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In vitro passage alters virulence, immune activation and proteomic profiles of Burkholderia pseudomallei
Taksaon Duangurai, Onrapak Reamtong, Amporn Rungruengkitkun, Varintip Srinon, Usa Boonyuen, Direk Limmathurotsakul, Narisara Chantratita, Pornpan Pumirat
Scientific Reports.2020;[Epub] CrossRef
Burkholderia pseudomallei Biofilm Promotes Adhesion, Internalization and Stimulates Proinflammatory Cytokines in Human Epithelial A549 Cells
Chanikarn Kunyanee, Watcharaporn Kamjumphol, Suwimol Taweechaisupapong, Sakawrat Kanthawong, Suwin Wongwajana, Surasak Wongratanacheewin, Chariya Hahnvajanawong, Sorujsiri Chareonsudjai, R. Mark Wooten
PLOS ONE.2016; 11(8): e0160741. CrossRef
A predicted cation transporter protein, BPSS1228, is involved in intracellular behaviour ofBurkholderia pseudomalleiin a human lung epithelial cell line (A549)
Teerasit Techawiwattanaboon, Sorujsiri Chareonsudjai, Craig Winstanley
FEMS Microbiology Letters.2016; 363(23): fnw259. CrossRef
Environmental Free-Living Amoebae Isolated from Soil in Khon Kaen, Thailand, Antagonize Burkholderia pseudomallei
Parumon Noinarin, Pisit Chareonsudjai, Pinich Wangsomnuk, Surasak Wongratanacheewin, Sorujsiri Chareonsudjai, William C. Nierman
PLOS ONE.2016; 11(11): e0167355. CrossRef
Mechanisms of Disease: Host-Pathogen Interactions between Burkholderia Species and Lung Epithelial Cells
Jonathan David, Rachel E. Bell, Graeme C. Clark
Frontiers in Cellular and Infection Microbiology.2015;[Epub] CrossRef

Note] Inhibition of quorum sensing in Pseudomonas aeruginosa by two herbal essential oils from Apiaceae family: Ehsan Sepahi , Saeed Tarighi , Farajollah Shahriari Ahmadi , Abdolreza Bagheri; J. Microbiol. 2015;53(2):176-180. Published online January 5, 2015; DOI: https://doi.org/10.1007/s12275-015-4203-8

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Ferula (Ferula asafoetida L.) and Dorema (Dorema aucheri Bioss.) both from Apiaceae family were tested for their antiquorum sensing (QS) activity against Pseudomonas aeruginosa. Both essential oils exhibited anti-QS activity at 25 μg/ml of concenteration. At this concenteration Ferula fully abolished and Dorema reduced the violacein production by C. violaceum. Pyocyanin, pyoverdine, elastase and biofilm production were decreased in Ferula oil treatments. Dorema oil reduced pyoverdine and elastase production, while pyocyanin and biofilm production were not affacted. Expresion analysis of QS-dependent genes confirmed our phenotypic data. Our data introduced native Dorema and Ferula plants as novel QS and virulence inhibitors.

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Research Support, U.S. Gov't, Non-P.H.S.

Description of Pseudomonas asuensis sp. nov. from biological soil crusts in the Colorado plateau, United States of America: Gundlapally Sathyanarayana Reddy , Ferran Garcia-Pichel; J. Microbiol. 2015;53(1):6-13. Published online January 4, 2015; DOI: https://doi.org/10.1007/s12275-015-4462-4

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A Gram-negative, aerobic, non spore-forming, non-motile, rod-shaped, yellow pigmented bacterium CP155-2T was isolated from a biological soil crusts sample collected in the Colorado plateau, USA and subjected to polyphasic taxonomic characterization. Strain CP155-2T contained summed feature 3 (C16:1ω5c/C16:1ω7c) and C18:1ω7c as major fatty acids and diphosphatidylglycerol (DPG) along with phosphatidylethanolamine (PE) and phosphatidylglycerol (PG) as major polar lipids. Based on these characteristics CP155-2T was assigned to the genus Pseudomonas. Phylogenetic analysis based on 16S rRNA gene sequence further confirmed the affiliation of CP155-2T to the genus Pseudomonas and showed a 16S rRNA gene sequence similarity of less than 98.7% with already described species of the genus. Pseudomonas luteola, Pseudomonas zeshuii, and Pseudomonas duriflava were identified as the closest species of the genus Pseudomonas with 16S rRNA gene sequence similarities of 98.7%, 98.6%, and 96.9%, respectively. The values for DNA–DNA relatedness between CP155-2T and Pseudomonas luteola and Pseudomonas zeshuii were 23% and 14% respectively a value below the 70% threshold value, indicating that strain CP155-2T belongs to a novel taxon of the genus Pseudomonas lineage. The novel taxon status was strengthened by a number of phenotypic differences wherein CP155-2T was positive for oxidase, negative for gelatin hydrolysis, could utilize D-cellobiose, D-raffinose, L-rhamnose, D-sorbitol but not L-aspartic acid and L-glutamic acid. Based on the collective differences strain CP155-2T exhibited, it was identified as a novel species and the name Pseudomonas asuensis sp. nov. was proposed. The type strain of Pseudomonas asuensis sp. nov. is CP155- 2T (DSM 17866T =ATCC BAA-1264T =JCM13501T =KCTC 32484T).

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Research Support, Non-U.S. Gov'ts

Pseudomonas aeruginosa-induced IL-1β Production is Inhibited by Sophora flavescens via the NF-κB/inflammasome Pathways: Jung-Hoon Lee , Heesung Shin , Yong-Jae Kim , Se-Hwan Paek , Shouguang Jin , Un-Hwan Ha; J. Microbiol. 2014;52(12):1044-1049. Published online November 29, 2014; DOI: https://doi.org/10.1007/s12275-014-4512-3

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The proinflammatory cytokine interleukin-1β plays an important role in protecting the host against airway infection; however, it can also trigger a massive influx of neutrophils into the airways, causing tissue damage. Anti-inflammatory treatments are particularly in demand for patients suffering from chronic inflammatory diseases. Sophora flavescens is a traditional herbal medicine used to reduce inflammation, but no study has examined its ability to block IL-1β production. Here, we show that S. flavescens reduced the Pseudomonas aeruginosa-induced expression of IL-1β by lung epithelial cells and macrophages. S. flavescens was also effective at reducing IL-1β production induced by either Staphylococcus aureus or phorbol 12-myristate 13-acetate, indicating that the effect is generalizable to diverse inflammatory stimuli. In addition, S. flavescens blocked the phosphorylation of IKKα/β, key upstream kinases involved in the degradation of IκBα, and the cleavage of caspase-1, a key component of the inflammasome. Thus, this study demonstrates that S. flavescens exerts its anti-inflammatory effects by blocking P. aeruginosa-mediated NF-κB/inflammasome activation and the subsequent production of IL-1β.

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The Role of Wheat Germ Agglutinin in the Attachment of Pseudomonas sp. WS32 to Wheat Root: Jian Zhang , Liyuan Meng , Yuanyuan Cao , Huiping Chang , Zhongyou Ma , Leni Sun , Ming Zhang , Xinyun Tang; J. Microbiol. 2014;52(12):1020-1024. Published online November 29, 2014; DOI: https://doi.org/10.1007/s12275-014-4089-x

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Wheat germ agglutinin (WGA), which is secreted on the surface of wheat root, has been defined as a protein that reversibly and non-enzymatically binds to specific carbohydrates. However, little attention has been paid to the function of WGA in the attachment of bacteria to their host plants. The aim of this study was to investigate the role of WGA in the attachment of Pseudomonas sp. WS32 to wheat roots. Wheat roots were initially treated with double-distilled water, WGA-H (WGA solution that was heated at 100°C for 15 min) and WGA, independently. Subsequently, the roots were coincubated with cell solutions (109 cells/ml). A dilution plate
method
using a solid nutrient medium was employed to determine the adsorption of WS32 to wheat roots. WGA was labeled with fluorescein isothiocyanate and detected using the fluorescent in situ hybridization (FISH) technique. The number of adsorptive WS32 cells on wheat roots was significantly increased when the wheat roots were pretreated with WGA, compared with the control treatment (p = 0.01). However, WGA-H failed to increase the amount of bacterial cells that attached to the wheat roots because of the loss of its physiological activity. The FISH assay also revealed that more cells adhered to WGA-treated wheat roots than to control or WGA-H-treated roots. The results indicated that WGA can mediate Pseudomonas strain WS32’s adherence to wheat seedling roots. The findings of this study provide a better understanding of the processes involved in plant-microbe interactions.

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Collaborative Impact of Wheat Germ Agglutinin and Bacillus subtilis on Growth, Water Balance, Photosynthetic Traits, Lignification and Suberinization of Wheat Seedlings
A. R. Lubyanova
Russian Journal of Plant Physiology.2025;[Epub] CrossRef
Isolation and purification of wheat germ agglutinin and analysis of its properties
Han Wang
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Identification of Proteolytic Bacteria from the Arctic Chukchi Sea Expedition Cruise and Characterization of Cold-active Proteases: Ha Ju Park , Yung Mi Lee , Sunghui Kim , Ah Ram Wi , Se Jong Han , Han-Woo Kim , Il-Chan Kim , Joung Han Yim , Dockyu Kim; J. Microbiol. 2014;52(10):825-833. Published online August 27, 2014; DOI: https://doi.org/10.1007/s12275-014-4226-6

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Following collection of seawater samples during an Arctic Chukchi Sea expedition cruise of the Korean icebreaker Araon in 2012, a total of 15,696 bacteria were randomly isolated from Marine Broth 2216 agar plates. Of these, 2,526 (16%) showed proteolytic activity and were identified as mainly Alteromonas (31%), Staphylococcus (27%), and Pseudoalteromonas (14%). Among the proteolytic strains, seven were selected based on their significant ability to grow and produce a halo on skim milk plates at low temperatures (<5°C) owing to cold-active proteases. These strains were affiliated with the genus Pseudoalteromonas and were divided into three groups based on phylogenetic analysis of the 16S rRNA genes. Profiling cell membrane fatty acids confirmed the 16S rRNA-based differentiation and revealed the accordance between the two analyses. Seven genes for serine protease precursors were amplified from the corresponding strains, and based on sequence similarities, these genes were divided into three groups that were identical to those identified by the 16S rRNA phylogenetic analysis. Three protease genes from the representative strains of each group were composed of 2,127–2,130 bp, encoding 708–709 amino acids, and these genes yielded products with calculated molecular weights of approximately 72.3–72.8 kDa. Amino acid sequence analysis suggested that the precursors are members of the subtilase serine endo- and exo-peptidase clan and contain four domains (signal peptide, N-terminal prosequence, catalytic domain, and two pre-peptidase C-terminal domains). Upon expression in E. coli, each recombinant protease exhibited proteolytic activity on zymogram gels.

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Diversity in the utilization of different molecular classes of dissolved organic matter by heterotrophic marine bacteria
Shira Givati, Elena Forchielli, Dikla Aharonovich, Noga Barak, Osnat Weissberg, Natalia Belkin, Eyal Rahav, Daniel Segrè, Daniel Sher, Jennifer F. Biddle
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Benthic bacteria and archaea in the North American Arctic reflect food supply regimes and impacts of coastal and riverine inputs
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Cold-Adapted Proteases: An Efficient and Energy-Saving Biocatalyst
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Description of Polaribacter batillariae sp. nov., Polaribacter cellanae sp. nov., and Polaribacter pectinis sp. nov., novel bacteria isolated from the gut of three types of South Korean shellfish
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Qi Wen, Xian-jie Liu, Wei-cong Zhu, Lu Li, Min-yi Li, Xuna-xian Peng, Hui Li
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Shengwei Hou, Mario López-Pérez, Ulrike Pfreundt, Natalia Belkin, Kurt Stüber, Bruno Huettel, Richard Reinhardt, Ilana Berman-Frank, Francisco Rodriguez-Valera, Wolfgang R Hess
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Pseudomonas aeruginosa MdaB and WrbA are Water-soluble Two-electron Quinone Oxidoreductases with the Potential to Defend against Oxidative Stress: Laura K Green , Anne C La Flamme , David F Ackerley; J. Microbiol. 2014;52(9):771-777. Published online August 2, 2014; DOI: https://doi.org/10.1007/s12275-014-4208-8

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Water-soluble quinone oxidoreductases capable of reducing quinone substrates via a concerted two-electron mechanism have been implicated in bacterial antioxidant defence. Twoelectron transfer avoids formation of dangerously reactive semi-quinone intermediates, moreover previous work in Pseudomonas putida indicated a direct protective effect for the quinols generated by an over-expressed oxidoreductase. Here, the Pseudomonas aeruginosa orthologs of five quinone oxidoreductases – MdaB, ChrR, WrbA, NfsB, and NQO1 – were tested for their possible role in defending P. aeruginosa against H2O2 challenge. In in vitro assays, each enzyme was shown to reduce quinone substrates with only minimal semiquinone formation. However, when each was individually over-expressed in P. aeruginosa no overt H2O2-protective phenotype was observed. It was shown that this was due to a masking effect of the P. aeruginosa catalase, KatA; in a katA mutant, H2O2 challenged strains over-expressing the WrbA and MdaB orthologs grew significantly better than the empty plasmid control. A growth advantage was also observed for H2O2 challenged P. putida strains over-expressing P. aeruginosa wrbA, mdaB or katA. Despite not conferring a growth advantage to wild type P. aeruginosa, it is possible that these quinone oxidoreductases defend against H2O2 toxicity at lower concentrations.

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Inhibitory Effects of 4-Hydroxy-2,5-Dimethyl-3(2H)-Furanone (HDMF) on Acyl-Homoserine Lactone-Mediated Virulence Factor Production and Biofilm Formation in Pseudomonas aeruginosa PAO1: Sung-Chan Choi , Can Zhang , Sooyoung Moon , Young-Sook Oh; J. Microbiol. 2014;52(9):734-742. Published online August 2, 2014; DOI: https://doi.org/10.1007/s12275-014-4060-x

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Abstract PDF

4-Hydroxy-2,5-dimethyl-3(2H)-furanone (HDMF), a nonhalogenated furanone found in a variety of fruits, has been shown to have antimicrobial activity. However, few studies have focused on its inhibitory effect on bacterial quorum sensing (QS) at levels below the non-inhibitory concentration. In this study, 0.1 μM HDMF decreased the production of QS signal molecules and inhibited QS-controlled biofilm formation by Pseudomonas aeruginosa PAO1 without causing growth inhibition. In the presence of 0.1 and 1.0 μM HDMF, biofilm production by PAO1 was reduced by 27.8 and 42.6%, respectively, compared to that by untreated control cells. HDMF (1.0 μM) also significantly affected virulence factor expression (regulated by the las, rhl, and pqs system), resulting in a significant reduction in the production of LasA protease (53.8%), rhamnolipid (40.9%), and pyocyanin (51.4%). This HDMF-dependent inhibition of virulence factor expression was overcome by increasing the levels of two QS signal molecules of P. aeruginosa, N-(3-oxo-dodecanoyl)-L-homoserine lactone and N-butyryl-L-homoserine lactone, suggesting reversible competitive inhibition between HDMF and these molecules. The results of this study indicate that HDMF has great potential as an inhibitor of QS, and that it may be of value as a therapeutic agent and in biofilm control, without increasing selective pressure for resistance development.

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Superinfection Exclusion Reveals Heteroimmunity between Pseudomonas aeruginosa Temperate Phages: In-Young Chung , Hee-Won Bae , Hye-Jung Jang , Bi-o Kim , You-Hee Cho; J. Microbiol. 2014;52(6):515-520. Published online May 29, 2014; DOI: https://doi.org/10.1007/s12275-014-4012-5

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Temperate siphophages (MP29, MP42, and MP48) were isolated from the culture supernatant of clinical Pseudomonas aeruginosa isolates. The complete nucleotide sequences and annotation of the phage genomes revealed the overall synteny to the known temperate P. aeruginosa phages such as MP22, D3112, and DMS3. Genome-level sequence analysis showed the conservation of both ends of the linear genome and the divergence at the previously identified dissimilarity regions (R1 to R9). Protein sequence alignment of the c repressor (ORF1) of each phage enabled us to divide the six phages into two groups: D3112 group (D3112, MP29, MP42, and MP48) and MP22 group (MP22 and DMS3). Superinfection exclusion was observed between the phages belonging to the same group, which was mediated by the specific interaction between the c repressor and the cognate operator. Based on these, we suggest that the temperate siphophages prevalent in the clinical strains of P. aeruginosa represent at least two distinct heteroimmunity groups.

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Validation Study

Comparison of JEV Neutralization Assay Using Pseudotyped JEV with the Conventional Plaque-Reduction Neutralization Test: Hee-Jung Lee , Kyung-Il Min , Ki Hoon Park , Hyo Jung Choi , Min-Kyoung Kim , Chi-Young Ahn , Young-Jin Hong , Young Bong Kim; J. Microbiol. 2014;52(5):435-440. Published online March 7, 2014; DOI: https://doi.org/10.1007/s12275-014-3529-y

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We previously reported the development of a neutralization assay system for evaluating Japanese Encephalitis Virus (JEV) neutralizing antibody (NAb) using pseudotyped-JEV (JEV- PV). JEV-PV-based neutralization assay offers several advan-tages compared with the current standard plaque-reduc-tion neutralization test (PRNT), including simplicity, safety, and speed. To evaluate the suitability of the JEV-PV assay as new replacement neutralization assay, we compared its repeatability, reproducibility, specificity, and correlated its results with those obtained using the PRNT. These analyses showed a close correlation between the results obtained with the JEV-PV assay and the PRNT, using the 50% plaque re-duction method as a standard for measuring NAb titers to JEV. The validation results met all analytical acceptance criteria. These results suggest that the JEV-PV assay could serve as a safe and simple method for measuring NAb titer against JEV and could be used as an alternative approach for assaying the potency of JEV neutralization.

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Reviews

REVIEW] Chronic Obstructive Pulmonary Disease (COPD): Evaluation From Clinical, Immunological and Bacterial Pathogenesis Perspectives: Daniel J. Hassett , Michael T. Borchers , Ralph J. Panos; J. Microbiol. 2014;52(3):211-226. Published online March 1, 2014; DOI: https://doi.org/10.1007/s12275-014-4068-2

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Chronic obstructive pulmonary disease (COPD), a disease manifested by significantly impaired airflow, afflicts ~14.2 million cases in the United States alone with an estimated 63 million people world-wide. Although there are a number of causes, the predominant cause is excessive tobacco smoke. In fact, in China, there have been estimates of 315,000,000 people that smoke. Other less frequent causes are associated with indirect cigarette smoke, air pollutants, biomass fuels, and genetic mutations. COPD is often associated with heart disease, lung cancer, osteoporosis and conditions can worsen in patients with sudden falls. COPD also affects both innate and adaptive immune processes. Cigarette smoke increases the expression of matrix metalloproteases and proinflammatory chemokines and increases lung titers of natural killer cells and neutrophils. Yet, neutrophil reactive oxygen species (ROS) mediated by the phagocytic respiratory burst and phagocytosis is impaired by nicotine. In contrast to innate immunity in COPD, dendritic cells represent leukocytes recruited to the lung that link the innate immune responses to adaptive immune responses by activating naïve T cells through antigen presentation. The autoimmune process that is also a significant part of inflammation associated with COPD. Moreover, coupled with restricted FEV1 values, are the prevalence of patients with single or multiple infections by bacteria, viruses and fungi. Finally, we focus on one of the more problematic infectious agents, the Gram-negative opportunistic pathogenic bacterium, Pseudomonas aeruginosa. Specifically, we delve into the development of highly problematic biofilm infections that are highly refractory to conventional antibiotic therapies in COPD. We offer a nonconventional, biocidal treatment that may be effective for COPD airway infections as well as with combinations of current antibiotic regimens for more effective treatment outcomes and relief for patients with COPD.

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REVIEW] Perturbation of Pulmonary Immune Functions by Carbon Nanotubes and Susceptibility to Microbial Infection: Brent E. Walling , Gee W. Lau; J. Microbiol. 2014;52(3):227-234. Published online March 1, 2014; DOI: https://doi.org/10.1007/s12275-014-3695-y

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Occupational and environmental pulmonary exposure to carbon nanotubes (CNT) is considered to be a health risk with a very low threshold of tolerance as determined by the United States Center for Disease Control. Immortalized airway epithelial cells exposed to CNTs show a diverse range of effects including reduced viability, impaired proliferation, and elevated reactive oxygen species generation. Additionally, CNTs inhibit internalization of targets in multiple macrophage cell lines. Mice and rats exposed to CNTs often develop pulmonary granulomas and fibrosis. Furthermore, CNTs have immunomodulatory properties in these animal models. CNTs themselves are proinflammatory and can exacerbate the allergic response. However, CNTs may also be immunosuppressive, both locally and systemically. Studies that examined the relationship of CNT exposure prior to pulmonary infection have reached different conclusions. In some cases, pre-exposure either had no effect or enhanced clearance of infections while other studies showed CNTs inhibited clearance. Interestingly, most studies exploring this relationship use pathogens which are not considered primary pulmonary pathogens. Moreover, harmony across studies is difficult as different types of CNTs have dissimilar biological effects. We used Pseudomonas aeruginosa as model pathogen to study how helical multi-walled carbon nanotubes (HCNTs) affected internalization and clearance of the pulmonary pathogen. The results showed that, although HCNTs can inhibit internalization through multiple processes, bacterial clearance was not altered, which was attributed to an enhanced inflammatory response caused by pre-exposure to HCNTs. We compare and contrast our findings in relation to other studies to gauge the modulation of pulmonary immune response by CNTs.

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Research Support, Non-U.S. Gov'ts

Molecular Characterization of the Alpha Subunit of Multicomponent Phenol Hydroxylase from 4-Chlorophenol-Degrading Pseudomonas sp. Strain PT3: Wael S. El-Sayed , Mohamed K. Ibrahim , Salama A. Ouf; J. Microbiol. 2014;52(1):13-19. Published online January 4, 2014; DOI: https://doi.org/10.1007/s12275-014-3250-x

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Multicomponent phenol hydroxylases (mPHs) are diiron enzymes that use molecular oxygen to hydroxylate a variety of phenolic compounds. The DNA sequence of the alpha subunit (large subunit) of mPH from 4-chlorophenol (4-CP)- degrading bacterial strain PT3 was determined. Strain PT3 was isolated from oil-contaminated soil samples adjacent to automobile workshops and oil stations after enrichment and establishment of a chlorophenol-degrading consortium. Strain PT3 was identified as a member of Pseudomonas sp. based on sequence analysis of the 16S rRNA gene fragment. The 4-CP catabolic pathway by strain PT3 was tentatively proposed to proceed via a meta-cleavage pathway after hydroxylation to the corresponding chlorocatechol. This hypothesis was supported by polymerase chain reaction (PCR) detection of the LmPH encoding sequence and UV/VIS spectrophotometric analysis of the culture filtrate showing accumulation of 5-chloro-2-hydroxymuconic semialdehyde (5-CHMS) with λmax 380. The detection of catabolic genes involved in 4-CP degradation by PCR showed the presence of both mPH and catechol 2,3-dioxygenase (C23DO). Nucleotide sequence analysis of the alpha subunit of mPH from strain PT3 revealed specific phylogenetic grouping to known mPH. The metal coordination encoding regions from strain PT3 were found to be conserved with those from the homologous dinuclear oxo-iron bacterial monooxygenases. Two DE(D)XRH motifs was detected in LmPH of strain PT3 within an approximate 100 amino acid interval, a typical arrangement characteristic of most known PHs.

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Trans-Membrane Transport of n-Octadecane by Pseudomonas sp. DG17: Fei Hua , Hong Qi Wang , Yi Li , Yi Cun Zhao; J. Microbiol. 2013;51(6):791-799. Published online December 19, 2013; DOI: https://doi.org/10.1007/s12275-013-3259-6

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The trans-membrane transport of hydrocarbons is an important and complex aspect of the process of biodegradation of hydrocarbons by microorganisms. The mechanism of transport of 14C n-octadecane by Pseudomonas sp. DG17, an alkane-degrading bacterium, was studied by the addition of ATP inhibitors and different substrate concentrations. When the concentration of n-octadecane was higher than 4.54 μmol/L, the transport of 14C n-octadecane was driven by a facilitated passive mechanism following the intra/extra substrate concentration gradient. However, when the cells were grown with a low concentration of the substrate, the cellular accumulation of n-octadecane, an energy-dependent process, was dramatically decreased by the presence of ATP inhibitors, and n-octadecane accumulation continually increased against its concentration gradient. Furthermore, the presence of non-labeled alkanes blocked 14C n-octadecane transport only in the induced cells, and the trans-membrane transport of n-octadecane was specific with an apparent dissociation constant Kt of 11.27 μmol/L and Vmax of 0.96 μmol/min/mg protein. The results indicated that the transmembrane transport of n-octadecane by Pseudomonas sp. DG17 was related to the substrate concentration and ATP.

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NOTE] Comparative Assessment of the Intracellular Survival of the Burkholderia pseudomallei bopC Mutant: Varintip Srinon , Sunsiree Muangman , Nithima Imyaem , Veerachat Muangsombut , Natalie R. Lazar Adler , Edouard E. Galyov , Sunee Korbsrisate; J. Microbiol. 2013;51(4):522-526. Published online August 30, 2013; DOI: https://doi.org/10.1007/s12275-013-2557-3

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Burkholderia pseudomallei, the causative agent of melioidosis, is a Gram-negative saprophytic bacterium capable of surviving within phagocytic cells. To assess the role of BopC (a type III secreted effector protein) in the pathogenesis of B. pseudomallei, a B. pseudomallei bopC mutant was used to infect J774A.1 macrophage-like cells. The bopC mutant showed significantly reduced intracellular survival in infected macrophages compared to wild-type B. pseudomallei. In addition, the bopC mutant displayed delayed escape from endocytic vesicles compared with the wild-type strain. This indicates that BopC is important, and at least in part, needed for intracellular survival of B. pseudomallei.

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pH Alkalinization by Chloroquine Suppresses Pathogenic Burkholderia Type 6 Secretion System 1 and Multinucleated Giant Cells
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Quantitative Proteomic Analysis of Burkholderia pseudomallei Bsa Type III Secretion System Effectors Using Hypersecreting Mutants
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