Journal of Microbiology

Review

Trans-acting regulators of ribonuclease activity: Jaejin Lee , Minho Lee , Kangseok Lee; J. Microbiol. 2021;59(4):341-359. Published online March 29, 2021; DOI: https://doi.org/10.1007/s12275-021-0650-6

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RNA metabolism needs to be tightly regulated in response to changes in cellular physiology. Ribonucleases (RNases) play an essential role in almost all aspects of RNA metabolism, including processing, degradation, and recycling of RNA molecules. Thus, living systems have evolved to regulate RNase activity at multiple levels, including transcription, post-transcription, post-translation, and cellular localization. In addition, various trans-acting regulators of RNase activity have been discovered in recent years. This review focuses on the physiological roles and underlying mechanisms of trans-acting regulators of RNase activity.

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Comparative Transcriptomic Analysis of Flagellar-Associated Genes in Salmonella Typhimurium and Its rnc Mutant
Seungmok Han, Ji-Won Byun, Minho Lee
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Relaxed Cleavage Specificity of Hyperactive Variants of Escherichia coli RNase E on RNA I
Dayeong Bae, Hana Hyeon, Eunkyoung Shin, Ji-Hyun Yeom, Kangseok Lee
Journal of Microbiology.2023; 61(2): 211. CrossRef
Regulator of RNase E activity modulates the pathogenicity of Salmonella Typhimurium
Jaejin Lee, Eunkyoung Shin, Ji-Hyun Yeom, Jaeyoung Park, Sunwoo Kim, Minho Lee, Kangseok Lee
Microbial Pathogenesis.2022; 165: 105460. CrossRef

Journal Articles

GABA-producing Lactobacillus plantarum inhibits metastatic properties and induces apoptosis of 5-FU-resistant colorectal cancer cells via GABAB receptor signaling: JaeJin An , Heon Seok , Eun-Mi Ha; J. Microbiol. 2021;59(2):202-216. Published online February 1, 2021; DOI: https://doi.org/10.1007/s12275-021-0562-5

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Abstract

5-Fluorouracil (5-FU) is an essential drug in systemic chemotherapy treatments for colorectal cancer (CRC). Despite the development of several treatment strategies over the past decades, the patient benefits of 5-FU-based therapies have been compromised by the development of chemoresistance. Differences in treatment responses among CRC patients may be due to genetic and epigenetic factors unique to individuals. Therefore, important factors for realizing personalized medicine are to accurately understand the causes and mechanisms of drug resistance to 5-FU-based therapies and to identify and validate prognostic biomarkers. Gut microbes that interact directly with the host contribute to human health and cancer control. Lactobacillus plantarum, in particular, has the potential to be a therapeutic agent by producing bioactive compounds that may benefit the host. Here, we investigated the gamma-aminobutyric acid (GABA) and GABAB receptor (GABABR)-dependent signaling pathway as a treatment option for 5-FU-resistant HT-29 cells. GABA-producing L. plantarum activates anti-proliferative, anti-migration, and anti-invasion effects against 5-FU-resistant HT-29 cells. The inhibitory effects of GABA-producing L. plantarum are mediated via GABABR. Activated GABABR induces apoptosis through the inhibition of cAMP-dependent signaling pathways and cellular inhibitor of apoptosis protein 2 (cIAP2) expression. Thus, the GABAergic system has potential in 5- FU-resistant HT-29 cells as a predictive biomarker. In addition, GABA-producing L. plantarum is promising as an adjuvant treatment for 5-FU-resistant CRC, and its intervention in neurobiological signaling imply new possibilities for chemoprevention and the treatment of colon cancer-related diseases.

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Epiberberine induced p53/p21-dependent G2/M cell cycle arrest and cell apoptosis in gastric cancer cells by activating γ-aminobutyric acid receptor- β3
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Biochemical Genetics.2024; 62(5): 4174. CrossRef
Intervention in gut microbiota increases intestinal γ-aminobutyric acid and alleviates anxiety behavior: a possible mechanism via the action on intestinal epithelial cells
Mion Ikegami, Hikari Narabayashi, Kazuaki Nakata, Miyu Yamashita, Yutaka Sugi, Yushiro Fuji, Hiroshi Matsufuji, Gaku Harata, Kazutoyo Yoda, Kenji Miyazawa, Yusuke Nakanishi, Kyoko Takahashi
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Sona Ciernikova, Aneta Sevcikova, Michal Mego
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Yihui Yang, Liwen Ren, Wan Li, Yizhi Zhang, Sen Zhang, Binbin Ge, Hong Yang, Guanhua Du, Bo Tang, Hongquan Wang, Jinhua Wang
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Biosynthesis of Gamma-Aminobutyric Acid (GABA) by Lactiplantibacillus plantarum in Fermented Food Production
Massimo Iorizzo, Gianluca Paventi, Catello Di Martino
Current Issues in Molecular Biology.2023; 46(1): 200. CrossRef
Lactobacillus plantarum Metabolites Elicit Anticancer Effects by Inhibiting Autophagy-Related Responses
Sihyun Jeong, Yuju Kim, Soyeong Park, Doyeon Lee, Juho Lee, Shwe Phyu Hlaing, Jin-Wook Yoo, Sang Hoon Rhee, Eunok Im
Molecules.2023; 28(4): 1890. CrossRef
Various Effects of the GABAergic System on Cancer: The Conditions and Specificities of its use in the Treatment of Some Cancers
Hossein Tahmasebi Dehkordi, Sorayya Ghasemi, Masoumeh Eliyasi Dashtaki
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The orchestra of human bacteriome by hormones
Arif Luqman
Microbial Pathogenesis.2023; 180: 106125. CrossRef
Gamma‐aminobutyric acid (GABA) can affect physiological processes in preimplantation embryos via GABAA and GABAB receptors
Veronika Kovaříková, Alexandra Špirková, Zuzana Šefčíková, Jozef Pisko, Laura Kalatová, Juraj Koppel, Dušan Fabian, Štefan Čikoš
Reproductive Medicine and Biology.2023;[Epub] CrossRef
Lactobacillus plantarum-derived metabolites sensitize the tumor-suppressive 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
Journal of Microbiology.2022; 60(1): 100. CrossRef
Engineered Bacteria-Based Living Materials for Biotherapeutic Applications
Rabia Omer, Muhammad Zubair Mohsin, Ali Mohsin, Bilal Sajid Mushtaq, Xumeng Huang, Meijin Guo, Yingping Zhuang, Jiaofang Huang
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Extracellular vesicles derived from Lactobacillus plantarum restore chemosensitivity through the PDK2-mediated glucose metabolic pathway in 5-FU-resistant colorectal cancer cells
JaeJin An, Eun-Mi Ha
Journal of Microbiology.2022; 60(7): 735. CrossRef
Role of lactobacillus strains in the management of colorectal cancer: An overview of recent advances
Elnaz Ghorbani, Amir Avan, Mikhail Ryzhikov, Gordon Ferns, Majid Khazaei, Saman Soleimanpour
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Gut Microbiota Eubacterium callanderi Exerts Anti-Colorectal Cancer Activity
Seoung Woo Ryu, Ji-Sun Kim, Byeong Seob Oh, Won Jung Choi, Seung Yeob Yu, Jeong Eun Bak, Seung-Hwan Park, Se Won Kang, Jiyoung Lee, Won Yong Jung, Jung-Sook Lee, Ju Huck Lee, Zhenjiang Zech Xu
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The intestinal microbiota in colorectal cancer metastasis – Passive observer or key player?
Meera Patel, Milly McAllister, Raghavendar Nagaraju, Sara Samir Foad Al Badran, Joanne Edwards, Andrew J. McBain, Jorge Barriuso, Omer Aziz
Critical Reviews in Oncology/Hematology.2022; 180: 103856. CrossRef
A comprehensive analysis of the microbiota composition and host driver gene mutations in colorectal cancer
Danping Yuan, Yong Tao, Haoyi Wang, Jiawei Wang, Yuepeng Cao, Wen Cao, Shou Pan, Zhaonan Yu
Investigational New Drugs.2022; 40(5): 884. CrossRef
Neurotransmitter signaling: a new frontier in colorectal cancer biology and treatment
Francesca Battaglin, Priya Jayachandran, Carly Strelez, Annika Lenz, Sandra Algaze, Shivani Soni, Jae Ho Lo, Yan Yang, Joshua Millstein, Wu Zhang, Evanthia T. Roussos Torres, Jean C. Shih, Shannon M. Mumenthaler, Josh Neman, Heinz-Josef Lenz
Oncogene.2022; 41(43): 4769. CrossRef
Probiotics and live biotherapeutic products aiming at cancer mitigation and patient recover
Zelinda Schemczssen-Graeff, Marcos Pileggi
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Gut microbiota: Linking nutrition and perinatal depression
Jia Song, Bi Zhou, Juntao Kan, Guangya Liu, Sheng Zhang, Liang Si, Xianping Zhang, Xue Yang, Junhua Ma, Junrui Cheng, Xiaobo Liu, Yongde Yang
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Exosome-Mediated Activation of Neuronal Cells Triggered by γ-Aminobutyric Acid (GABA)
Ryo Inotsuka, Miyako Udono, Atsushi Yamatsu, Mujo Kim, Yoshinori Katakura
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Restoring HOXD10 Exhibits Therapeutic Potential for Ameliorating Malignant Progression and 5-Fluorouracil Resistance in Colorectal Cancer
Weijie Pan, Kaijing Wang, Jiayong Li, Hanhua Li, Yuchan Cai, Min Zhang, Aili Wang, Yazhou Wu, Wei Gao, Wenhao Weng
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In vitro disinfection efficacy and clinical protective effects of common disinfectants against acute hepatopancreatic necrosis disease (AHPND)-causing Vibrio isolates in Pacific white shrimp Penaeus vannamei: Peizhuo Zou , Qian Yang , Hailiang Wang , Guosi Xie , Zhi Cao , Xing Chen , Wen Gao , Jie Huang; J. Microbiol. 2020;58(8):675-686. Published online July 27, 2020; DOI: https://doi.org/10.1007/s12275-020-9537-1

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Abstract

Acute hepatopancreatic necrosis disease (AHPND) is one of the most significant bacterial diseases in global shrimp culture, causing severe economic losses. In the present study, we carried out in vitro antimicrobial tests to investigate the disinfection efficacy of 14 common disinfectants toward different AHPND-causing Vibrio spp., including eight isolates of V. parahaemolyticus, four isolates of V. campbellii, and one isolate of V. owensii. Polyhexamethylene biguanidine hydrochloride (PHMB) was revealed to possess the strongest inhibitory activity. Through analyzing and evaluating the results of antimicrobial tests and acute toxicity test, we selected PHMB and hydrogen peroxide (H2O2) for further clinical protection test. Clinical manifestations indicated that both PHMB (2 mg/L and 4 mg/L) and H2O2 (12 mg/L) could effectively protect juvenile Penaeus vannamei from the infection of V. parahaemolyticus isolate Vp362 at 106 CFU/ml, and the survival rate was over 80%. When the bacterial concentration was reduced to 105 CFU/ml, 104 CFU/ml, and 103 CFU/ml, the survival rate after treated by 1 mg/L PHMB was 64.44%, 93.33%, and 100%, respectively. According to the results, PHMB and H2O2 showed a lower toxicity while a better protection activity, particularly against a lower concentration of the pathogens. Therefore, these two disinfectants are proved to be promising disinfectants that can be applied to prevent and control AHPND in shrimp culture. Moreover, the methods of this study also provided valuable information for the prevention of other important bacterial diseases and suggested a reliable means for screening potential drugs in aquaculture.

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Basigin binds bacteria and activates Dorsal signaling to promote antibacterial defense in Penaeus vannamei
Linwei Yang, Zi-ang Wang, Yushi Gan, Hongliang Zuo, Hengwei Deng, Shaoping Weng, Jianguo He, Xiaopeng Xu
Fish & Shellfish Immunology.2023; 142: 109123. CrossRef
Clinical protective effects of polyhexamethylene biguanide hydrochloride (PHMB) against Vibrio parahaemolyticus causing translucent post-larvae disease (VTPD) in Penaeus vannamei
Tianchang Jia, Tingting Xu, Jitao Xia, Shuang Liu, Wenqiang Li, Ruidong Xu, Jie Kong, Qingli Zhang
Journal of Invertebrate Pathology.2023; 201: 108002. CrossRef

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

Detection of Inhibitors of Phenotypically Drug-tolerant Mycobacterium tuberculosis Using an In Vitro Bactericidal Screen: Ian M. Bassett , Shichun Lun , William R. Bishai , Haidan Guo , Joanna R. Kirman , Mudassar Altaf , Ronan F. O’Toole; J. Microbiol. 2013;51(5):651-658. Published online June 25, 2013; DOI: https://doi.org/10.1007/s12275-013-3099-4

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Abstract

Many whole cell screens of chemical libraries currently in use are based on inhibition of bacterial growth. The goal of this study was to develop a chemical library screening model that enabled detection of compounds that are active against drug-tolerant non-growing cultures of Mycobacterium tuberculosis. An in vitro model of low metabolically active mycobacteria was established with 8 and 30 day old cultures of M. smegmatis and M. tuberculosis, respectively. Reduction of resazurin was used as a measure of viability and the assay was applied in screens of chemical libraries for bactericidal compounds. The model provided cells that were phenotypically-resilient to killing by first and second-line clinical drugs including rifampicin. Screening against chemical libraries identified proteasome inhibitors, NSC310551 and NSC321206, and a structurally-related series of thiosemicarbazones, as having potent killing activity towards aged cultures. The inhibitors were confirmed as active against virulent M. tuberculosis strains including multi- and extensively-drug resistant clinical isolates. Our library screen enabled detection of compounds with a potent level of bactericidal activity towards phenotypically drug-tolerant cultures of M. tuberculosis.

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Cytotoxicity and activity of thiosemicarbazones and semicarbazones in Mycobacterium tuberculosis: a systematic review
Ana Paula Desiree de Oliveira, Eloísa Gibin Sampiron, Jean Eduardo Meneguello, Andressa Lorena Ieque, Katiany Rizzieri Caleffi Ferracioli, Rosilene Fressatti Cardoso, Fábio Vandresen, Regiane Bertin de Lima Scodro
Cuadernos de Educación y Desarrollo.2024; 16(6): e4683. CrossRef
High-Throughput Screening of Natural Product and Synthetic Molecule Libraries for Antibacterial Drug Discovery
Navid J. Ayon
Metabolites.2023; 13(5): 625. CrossRef
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Summaya Perveen, Rashmi Sharma
Biochemical Pharmacology.2022; 197: 114906. CrossRef
Anti-tuberculosis treatment strategies and drug development: challenges and priorities
Véronique A. Dartois, Eric J. Rubin
Nature Reviews Microbiology.2022; 20(11): 685. CrossRef
Identification of small molecules targeting homoserine acetyl transferase from Mycobacterium tuberculosis and Staphylococcus aureus
Deepika Chaudhary, Avantika Singh, Mardiana Marzuki, Abhirupa Ghosh, Saqib Kidwai, Tannu Priya Gosain, Kiran Chawla, Sonu Kumar Gupta, Nisheeth Agarwal, Sudipto Saha, Yashwant Kumar, Krishan Gopal Thakur, Amit Singhal, Ramandeep Singh
Scientific Reports.2022;[Epub] CrossRef
In vitro drug discovery models for Mycobacterium tuberculosis relevant for host infection
Tanya Parish
Expert Opinion on Drug Discovery.2020; 15(3): 349. CrossRef
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Soma Mandal, Samuel Njikan, Anuradha Kumar, Julie V. Early, Tanya Parish
Microbiology .2019; 165(5): 492. CrossRef
Selective Killing of Dormant Mycobacterium tuberculosis by Marine Natural Products
Carolina Rodrigues Felix, Rashmi Gupta, Sandra Geden, Jill Roberts, Priscilla Winder, Shirley A. Pomponi, Maria Cristina Diaz, John K. Reed, Amy E. Wright, Kyle H. Rohde
Antimicrobial Agents and Chemotherapy.2017;[Epub] CrossRef
Targeting Phenotypically TolerantMycobacterium tuberculosis
Ben Gold, Carl Nathan, William R. Jacobs Jr., Helen McShane, Valerie Mizrahi, Ian M. Orme
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Optimization and Evaluation of 5-Styryl-Oxathiazol-2-oneMycobacterium tuberculosisProteasome Inhibitors as Potential Antitubercular Agents
Francesco Russo, Johan Gising, Linda Åkerbladh, Annette K. Roos, Agata Naworyta, Sherry L. Mowbray, Anders Sokolowski, Ian Henderson, Torey Alling, Mai A. Bailey, Megan Files, Tanya Parish, Anders Karlén, Mats Larhed
ChemistryOpen.2015; 4(3): 342. CrossRef

Bacteria-Based In Vivo Peptide Library Screening Using Biopanning Approach: Ji-Hyeon Choi , Sang-Hyun Park; J. Microbiol. 2011;49(5):847-851. Published online November 9, 2011; DOI: https://doi.org/10.1007/s12275-011-1405-6

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Abstract: Traditionally, library screening has been performed to identify biologically active agents including small molecules or peptides that inhibit target proteins or molecules with therapeutic interests. Due to its chemical nature, library screening is usually performed under in vitro environments using purified proteins and molecules. However, active agents identified from in vitro screenings often fail to exhibit biological activities in cells. To overcome this inherent limitation, we have developed an in vivo peptide library screening system that allows for the identification of dissociative inhibitors of protein interactions of interest. The screening is based on the reconstitution of the cI repressor from bacteriophage lambda with high-density expression peptide library and is entirely performed in bacteria cells. Furthermore, to enhance the efficacy and sensitivity of the screening, a multiple-round biopanning approach was employed for amplification and enrichment of positive peptides. Overall, this in vivo screening should provide a fast and efficient tool for identification of biologically active peptide molecules against target protein assembly.

Identification of the Genes Involved in 1-Deoxynojirimycin Synthesis in Bacillus subtilis MORI 3K-85: Kyung-Don Kang , Yong Seok Cho , Ji Hye Song , Young Shik Park , Jae Yeon Lee , Kyo Yeol Hwang , Sang Ki Rhee , Ji Hyung Chung , Ohsuk Kwon , Su-Il Seong; J. Microbiol. 2011;49(3):431-440. Published online June 30, 2011; DOI: https://doi.org/10.1007/s12275-011-1238-3

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Abstract: 1-Deoxynojirimycin (DNJ), a D-glucose analogue with a nitrogen atom substituting for the ring oxygen, is a strong inhibitor of intestinal α-glucosidase. DNJ has several promising biological activities, including its antidiabetic, antitumor, and antiviral activities. Nevertheless, only limited amounts of DNJ are available because it can only be extracted from some higher plants, including the mulberry tree, or purified from the culture broth of several types of soil bacteria, such as Streptomyces sp. and Bacillus sp. In our previous study, a DNJ-producing bacterium, Bacillus subtilis MORI, was isolated from the traditional Korean fermented food Chungkookjang. In the present study, we report the identification of the DNJ biosynthetic genes in B. subtilis MORI 3K-85 strain, a DNJ-overproducing derivate of the B. subtilis MORI strain generated by γ-irradiation. The genomic DNA library of B. subtilis MORI 3K-85 was constructed in Escherichia coli, and clones showing α-glucosidase inhibition activity were selected. After DNA sequencing and a series of subcloning, we were able to identify a putative operon which consists of gabT1, yktc1, and gutB1 genes predicted to encode putative transaminase, phosphatase, and oxidoreductase, respectively. When a recombinant plasmid containing this operon sequence was transformed into an E. coli strain, the resulting transformant was able to produce DNJ into the culture medium. Our results indicate that the gabT1, yktc1, and gutB1 genes are involved in the DNJ biosynthetic pathway in B. subtilis MORI, suggesting the possibility of employing these genes to establish a large-scale microbial DNJ overproduction system through genetic engineering and process optimization.

Molecular Cloning, Purification, and Characterization of a Superoxide Dismutase from a Fast-Growing Mycobacterium sp. Strain JC1 DSM 3803: Ji-Sun Nam , Jee-Hyun Yoon , Hyun-Il Lee , Si Wouk Kim , Young-Tae Ro; J. Microbiol. 2011;49(3):399-406. Published online June 30, 2011; DOI: https://doi.org/10.1007/s12275-011-1046-9

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Abstract: A cytosolic superoxide dismutase (SOD) was purified and characterized from a fast-growing Mycobacterium sp. strain JC1 DSM 3803 grown on methanol. The native molecular weight of the purified SOD was estimated to be 48 kDa. SDS-PAGE revealed a subunit of 23 kDa, indicating that the enzyme is a homodimer. The enzyme activity was inhibited by H2O2 and azide. The purified SOD contained 1.12 and 0.56 g-atom of Mn and Fe per mol of enzyme, respectively, suggesting that it may be a Fe/Mn cambialistic SOD. The apo-SOD reconstitution study revealed that Mn salts were more specific than Fe salts in the SOD activity. The gene encoding the SOD was identified from the JC1 cosmid genomic library by PCR screening protocol. The cloned gene, sodA, had an open reading frame (ORF) of 624 nt, encoding a protein with a calculated molecular weight of 22,930 Da and pI of 5.33. The deduced SodA sequence exhibited 97.6% identity with that of Mycobacterium fortuitum Mn-SOD and clustered with other mycobacterial Mn-SODs. A webtool analysis on the basis of SOD sequence and structure homologies predicted the SOD as a tetrameric Mn-SOD, suggesting that the protein is a dimeric Mn-SOD having tetramer-specific sequence and structure characteristics.

NOTE] Development of a High-Throughput Screening Method for Recombinant Escherichia coli with Intracellular Dextransucrase Activity: So-Ra Lee , Ah-Rum Yi , Hong-Gyun Lee , Myoung-Uoon Jang , Jung-Mi Park , Nam Soo Han , Tae-Jip Kim; J. Microbiol. 2011;49(2):320-323. Published online May 3, 2011; DOI: https://doi.org/10.1007/s12275-011-1078-1

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Abstract: To efficiently engineer intracellular dextransucrase (DSase) expression in Escherichia coli, a high-throughput screening method was developed based on the polymer-forming activity of the enzyme. Recombinant E. coli containing the Leuconostoc citreum DSase (LcDS) gene was grown on Luria-Bertani agar plates, containing 2% sucrose, at 37°C for 8 h. The plates were then evenly overlaid with 0.6% soft agar, containing 1.2 mg/ml D-cycloserine, and incubated at 30°C to allow gradual cell disruption until a dextran polymer grew through the overlaid layer. A significant correlation between dextran size and enzyme activity was established and applied for screening truncated mutants with LcDS activity.

Bacterial Model System for Screening and Determining Optimal Concentration of Anti-caries Natural Extracts: Min Jung Kim , Chun Sung Kim , Jae-Yoon Park , Soon-Nang Park , So Young Yoo , Sook-Young Lee , Joong-Ki Kook; J. Microbiol. 2011;49(1):165-168. Published online March 3, 2011; DOI: https://doi.org/10.1007/s12275-011-1018-0

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Abstract: In general, an antimicrobial test for screening anti-caries natural extracts was performed by measuring the minimum bactericidal concentration (MBC) against the type strains of mutans streptococci. However, it is unclear if the antimicrobial efficiency of natural extracts on the type strains of mutans streptococci is the same on the clinical strains. In this study, we introduced a bacterial model system for the screening of anti-caries and determining the optimal concentration of them to develop oral hygiene products for Korean populations.

Isolation and Characterization of Marine Pigmented Bacteria from Norwegian Coastal Waters and Screening for Carotenoids with UVA-Blue Light Absorbing Properties: Marit H. Stafsnes , Kjell D Josefsen , Geir Kildahl-Andersen , Svein Valla , Trond E. Ellingsen , Per Bruheim; J. Microbiol. 2010;48(1):16-23. Published online March 11, 2010; DOI: https://doi.org/10.1007/s12275-009-0118-6

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Abstract: Microbial culture collections are important resources for isolation of natural compounds with novel properties. In this study, a culture collection of around 1,500 pigmented heterotrophic bacteria was established. The bacteria were isolated from the sea surface microlayer at different sampling sites along the mid-part of the Norwegian coast. The bacterial isolates produced pigments of various coloration (e.g. golden, yellow, red, pink and orange). Methanol extracts of sixteen isolates were characterized with LC-Diodearray-TOF mass spectrometry analysis. The number of pigments per isolate varied considerably, and a tentative identification of the pigments was performed based on UV-absorbance profile and molecular formula assignation based on the accurate mass determination. The LC-MS analyses evealed that most of the pigments probably were carotenoids. Furthermore, we developed a high throughput LC-MS method for characterization and screening of a larger sub-fraction (300 isolates) of the culture collection. The aim was to screen and identify bacterial isolates producing carotenoids that absorb light in the UVA-Blue light. Six of the bacterial strains were selected for detailed investigation, including 16s rRNA sequencing, preparative HPLC for purification of major carotenoids and subsequent structural elucidation with NMR. Among the identified carotenoids were zeaxanthin, nostoxanthin and sarcinaxanthin, some with novel glycosylation patterns.

Characterization of a Novel β-Glucosidase-Like Activity from a Soil Metagenome: Chengjian Jiang , Gefei Ma , Shuangxi Li , Tingting Hu , Zhiqun Che , Peihong Shen , Bing Yan , Bo Wu; J. Microbiol. 2009;47(5):542-548. Published online October 24, 2009; DOI: https://doi.org/10.1007/s12275-009-0024-y

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Abstract: We report the cloning of a novel β-glucosidase-like gene by function-based screening of a metagenomic library from uncultured soil microorganisms. The gene was named bgl1C and has an open reading frame of 1,443 base pairs. It encodes a 481 amino acid polypeptide with a predicted molecular mass of about 57.8 kDa. The deduced amino acid sequence did not show any homology with known β-glucosidases. The putative β-glucosidase gene was subcloned into the pETBlue-2 vector and overexpressed in E. coli Tuner (DE3) pLacІ; the recombinant protein was purified to homogeneity. Functional characterization with a high performance liquid chromatography method demonstrated that the recombinant Bgl1C protein hydrolyzed D-glucosyl-β-(1-4)-D-glucose to glucose. The maximum activity for Bgl1C protein occurred at pH 8.0 and 42°C using p-nitrophenyl-β-D-glucoside as the substrate. A CaCl2 concentration of 1 mM was required for optimal activity. The putative β-glucosidase had an apparent Km value of 0.19 mM, a Vmax value of 4.75 U/mg and a kcat value of 316.7/min under the optimal reaction conditions. The biochemical characterization of Bgl1C has enlarged our understanding of the novel enzymes that can be isolated from the soil metagenome.

Screening of Genes Related to Methylglyoxal Susceptibility: Insook Kim , Joonho Kim , Bumchan Min , Changhan Lee , Chankyu Park; J. Microbiol. 2007;45(4):339-343.; DOI: https://doi.org/2563 [pii]

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Abstract: Methylglyoxal (MG) is a reactive metabolite known to accumulate in certain physiological conditions. We attempted to isolate genes associated with this metabolite by genome-wide mutagenesis with TnphoA derivative. After screening on methylglyoxal-containing plate, we obtained insertions in three different genes, ydbD, yjjQ, and yqiI, which gave rise to reproducible MG-sensitive phenotypes in glyoxalase-deficient strain. In addition to its MG sensitivity, the insertion in yqiI exhibited an impaired motility resulting from a reduced flagellar expression.

Identification and characterization of pH-regulated genes in saccharomyces cerevisiae: Hong, Sung Ki , Choi, Eui Yul; J. Microbiol. 1996;34(4):327-333.

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Abstract: Yeast, like many other microbes, encounters large variations in ambient pH in their natural environments. Microorganisms capable of growing over a wide pH range require a versatile, efficient pH homeostatic mechanism protecting intracellular processes against extremes of pH. In several organisms, fusions to the bacterial lacZ gene have been extremely useful for the identification of genes expressed at different time during the life cycle or under different growth conditions. In this study, using the lacZ gene screening system, we surveyed a large number of yeast strains with lacZ insertion to identify genes regulated by pH. A yeast genomic library was constructed and inserted with lacZ by a shuttle mutagenesis procedure. The yeast transformants were individually picked up with a toothpick, replica-plated, and grown in alkaline pH medium. Among the 35,000 colonies screened, 10 candidate strains were identified initially by the β-gal assay. We finally confirmed two yeast strains carrying the genes whose expression are strictly dependent on pH of growth medium. One of the fusions showing a 10-fodl induction in expression level in response to alkali pH was selected and further characterized. The pH-regulated gene was cloned by inverse PCR and a partial sequence of the gene was determined. Identification and characterization of the gene is currently under investigation.

Genomic Organization of Penicillium chrysogenum chs4, a Class III Chitin Synthase Gene: Yoon-Dong Park , Myung-Sook Lee , Ji-Hoon Kim , Jun Namgung , Bum Chan Park , Kyung Sook Bae , Hee-Moon Park; J. Microbiol. 2000;38(4):230-238.

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Abstract: Class III chitin synthases in filamentous fungi are important for hyphal growth and differentiation of several filamentous fungi. A genomic clone containing the full gene encoding Chs4, a class III chitin synthase in Penicillium chrysogenum, was cloned by PCR screening and colony hybridization from the genomic library. Nucleotide sequence analysis and transcript mapping of chs4 revealed an open reading frame (ORF) that consisted of 5 exons and 4 introns and encoded a putative protein of 915 amino acids. Nucleotide sequence analysis of the 5?lanking region of the ORF revealed a potential TATA box and several binding sites for transcription activators. The putative transcription initiation site at ?6 position was identified by primer extension and the expression of the chs4 during the vegetative growth was confirmed by Northern blot analysis. Amino acid sequence analysis of the Chs4 revealed at least 5 transmembrane helices and several sites for post-translational modifications. Comparison of the amino acid sequence of Chs4 with those of other fungi showed a close relationship between P. chr ysogenum and genus Aspergillus.

Assessment of Baird-Parker Agar as Screening Test for Determination of Staphylococcus aureus in Poultry Meat: Rosa Capita , Carlos Alonso-Calleja , Benito Moreno , Maria del Camino Garcia-Fernandez; J. Microbiol. 2001;39(4):321-325.

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Abstract: Baird-Parker agar with egg yolk/tellurite emulsion (BPA) is widely accepted as a medium for the enumeration of Staphylococcus aureus in foods. However, it is not completely selective and colonies of other genera or species could be similar to those of Staphylococcus aureus. Moreover, the strains of Staphylococcus aureus that are lecithinase negative could go unnoticed. Both facts could affect the counts. The aim of this study was to determine whether the enumeration of the colonies with the typical morphology of Staphylococcus aureus on BPA is sufficient to quantify this species in poultry meat. Forty chicken carcasses were tested for Staphylococcus aureus by surface plating using BPA. Results indicate that the predictive value of the morphology of the colonies on BPA is 85.71% and 68.42% for typical and atypical colonies of Staphylococcus aureus, respectively. However, Staphylococcus aureus counts (after identification) and counts of typical colonies did not show any significant differences (P>0.05) and are significantly (P<0.001) correlated (r = 0.996). These results suggest that, for screening purposes, enumeration of Staphylococcus aureus from poultry meat does not require any identification of strains, resulting in a saving of time and money.

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