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- Incomplete autophagy promotes the replication of Mycoplasma hyopneumoniae
- Zhaodi Wang† , Yukang Wen† , Bingqian Zhou , Yaqin Tian , Yaru Ning , Honglei Ding
- J. Microbiol. 2021;59(8):782-792. Published online July 5, 2021
- DOI: https://doi.org/10.1007/s12275-021-1232-3
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Abstract
- Autophagy is an important cellular homeostatic mechanism for recycling of degradative proteins and damaged organelles. Autophagy has been shown to play an important role in cellular responses to bacteria and bacterial replication. However, the role of autophagy in Mycoplasma hyopneumoniae infection and the pathogenic mechanism is not well characterized. In this study, we showed that M. hyopneumoniae infection significantly increases the number of autophagic vacuoles in host cells. Further, we found significantly enhanced expressions of autophagy marker proteins (LC3-II, ATG5, and Beclin 1) in M. hyopneumoniae-infected cells. Moreover, immunofluorescence analysis showed colocalization of P97 protein with LC3 during M. hyopneumoniae infection. Interestingly, autophagic flux marker, p62, accumulated with the induction of infection. Conversely, the levels of p62 and LC3-II were decreased after treatment with 3-MA, inhibiting the formation of autophagosomes, during infection. In addition, accumulation of autophagosomes promoted the expression of P97 protein and the survival of M. hyopneumoniae in PK- 15 cells, as the replication of M. hyopneumoniae was downregulated by adding 3-MA. Collectively, these findings provide strong evidence that M. hyopneumoniae induces incomplete autophagy, which in turn enhances its reproduction in host cells. These findings provide novel insights into the interaction of M. hyopneumoniae and host.
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Citations
Citations to this article as recorded by
- Research Progress on Immune Evasion of Mycoplasma hyopneumoniae
Bin Jiang, Ying Zhang, Gaojian Li, Yanping Quan, Jianhong Shu, Huapeng Feng, Yulong He
Microorganisms.2024; 12(7): 1439. CrossRef - The Role of Pyroptosis and Autophagy in Ischemia Reperfusion Injury
Huijie Zhao, Yihan Yang, Xinya Si, Huiyang Liu, Honggang Wang
Biomolecules.2022; 12(7): 1010. CrossRef - Mycoplasma bovis inhibits autophagy in bovine mammary epithelial cells via a PTEN/PI3K-Akt-mTOR-dependent pathway
Maolin Xu, Yang Liu, Tuerdi Mayinuer, Yushan Lin, Yue Wang, Jian Gao, Dong Wang, John P. Kastelic, Bo Han
Frontiers in Microbiology.2022;[Epub] CrossRef - Incomplete autophagy promotes the proliferation of Mycoplasma hyopneumoniae through the JNK and Akt pathways in porcine alveolar macrophages
Yukang Wen, Zhengkun Chen, Yaqin Tian, Mei Yang, Qingshuang Dong, Yujiao Yang, Honglei Ding
Veterinary Research.2022;[Epub] CrossRef
- Research Progress on Immune Evasion of Mycoplasma hyopneumoniae
- Type 2 human papillomavirus E7 attenuates E-cadherin expression in human keratinocytes
- Ji Young Song , Young Min Park , Soon Yong Choi
- J. Microbiol. 2021;59(6):616-625. Published online March 29, 2021
- DOI: https://doi.org/10.1007/s12275-021-0690-y
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Abstract
- Human papillomaviruses (HPVs) are known to utilize the down-regulation of epithelial (E)-cadherin, a major component of adherens junctions of keratinocytes, to evade host immune surveillance in high-risk group. However, the effects of HPV on the function of E-cadherin in low-risk groups remain unknown. We investigated whether type 2 HPV (HPV- 2) E7 could induce alterations in E-cadherin expression in transiently transfected keratinocytes and cell lines expressing HPV-2 E7. To examine the expression pattern of E-cadherin in cutaneous warts and normal skin samples, immunohistochemical analysis was performed. Quantitative real-time polymerase chain reactions, luciferase assays, western blot, immunocytochemistry, and electron microscopy were used to evaluate the mRNA and protein expression levels of Ecadherin in normal human epidermal keratinocytes transfected with HPV-2 E7 plasmid DNA or E7-specific siRNA and in E7-expressing cell lines. E-cadherin expression levels in HPV-2 positive cutaneous warts were significantly decreased compared to those in normal skin (p < 0.05). Similarly, the mRNA and protein expression levels of E-cadherin in E7 transiently transfected cells were significantly decreased compared to those in empty vector-transfected cells. The decreases were restored by transfection with E7-specific siRNA (p < 0.05). Likewise, cell lines expressing E7 showed a decreased expression of E-cadherin. When the cells were cultured in low attachment plates, cell-to-cell aggregation was inhibited. Taken together, our data suggest that HPV-2 E7, the causative agent of cutaneous warts, could mediate the transcriptional repression of E-cadherin.
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Citations
Citations to this article as recorded by- The NLRP3 inflammasome in viral infection (Review)
Qiaoli Zheng, Chunting Hua, Qichang Liang, Hao Cheng
Molecular Medicine Reports.2023;[Epub] CrossRef
- The NLRP3 inflammasome in viral infection (Review)
- Characterization of a novel dsRNA mycovirus of Trichoderma atroviride NFCF377 reveals a member of “Fusagraviridae” with changes in antifungal activity of the host fungus
- Jeesun Chun , Byeonghak Na , Dae-Hyuk Kim
- J. Microbiol. 2020;58(12):1046-1053. Published online October 23, 2020
- DOI: https://doi.org/10.1007/s12275-020-0380-1
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Abstract
- Trichoderma atroviride is a common fungus found in various ecosystems that shows mycoparasitic ability on other fungi. A novel dsRNA virus was isolated from T. atroviride NFCF377 strain and its molecular features were analyzed. The viral genome consists of a single segmented double-stranded RNA and is 9,584 bp in length, with two discontinuous open reading frames (ORF1 and ORF2). A mycoviral structural protein and an RNA-dependent RNA polymerase (RdRp) are encoded by ORF1 and ORF2, respectively, between which is found a canonical shifty heptameric signal motif (AAAAAAC) followed by an RNA pseudoknot. Analysis of sequence similarity and phylogeny showed that it is closely related to members of the proposed family “Fusagraviridae”, with a highest similarity to the Trichoderma atroviride mycovirus 1 (TaMV1). Although the sequence similarity of deduced amino acid to TaMV1 was evident, sequence deviations were distinctive at untranslated regions (UTRs) due to the extended size. Thus, we inferred this dsRNA to be a different strain of Trichoderma atroviride mycovirus 1 (TaMV1-NFCF377). Electron microscopy image exhibited an icosahedral viral particle of 40 nm diameter. Virus-cured isogenic isolates were generated and no differences in growth rate, colony morphology, or conidia production were observed between virus-infected and virus-cured strains. However, culture filtrates of TaMV1- NFCF377-infected strain showed enhanced antifungal activity against the plant pathogen Rhizoctonia solani but not to edible mushroom Pleurotus ostreatus. These results suggested that TaMV1-NFCF377 affected the metabolism of the fungal host to potentiate antifungal compounds against a plant pathogen, but this enhanced antifungal activity appeared to be species-specific.
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Citations
Citations to this article as recorded by- Co-infection with two novel mycoviruses affects the biocontrol activity of Trichoderma polysporum
Jeesun Chun, Hae-Ryeong Yoon, Sei-Jin Lee, Dae-Hyuk Kim
Biological Control.2024; 188: 105440. CrossRef -
An Outstandingly Rare Occurrence of Mycoviruses in Soil Strains of the Plant-Beneficial Fungi from the Genus
Trichoderma
and a Novel
Polymycoviridae
Isolate
Chenchen Liu, Xiliang Jiang, Zhaoyan Tan, Rongqun Wang, Qiaoxia Shang, Hongrui Li, Shujin Xu, Miguel A. Aranda, Beilei Wu, Lea Atanasova
Microbiology Spectrum.2023;[Epub] CrossRef - Sixteen Novel Mycoviruses Containing Positive Single-Stranded RNA, Double-Stranded RNA, and Negative Single-Stranded RNA Genomes Co-Infect a Single Strain of Rhizoctonia zeae
Siwei Li, Zhihao Ma, Xinyi Zhang, Yibo Cai, Chenggui Han, Xuehong Wu
Journal of Fungi.2023; 10(1): 30. CrossRef - Trichoderma – genomes and genomics as treasure troves for research towards biology, biotechnology and agriculture
Miriam Schalamun, Monika Schmoll
Frontiers in Fungal Biology.2022;[Epub] CrossRef - A Transfectable Fusagravirus from a Japanese Strain of Cryphonectria carpinicola with Spherical Particles
Subha Das, Sakae Hisano, Ana Eusebio-Cope, Hideki Kondo, Nobuhiro Suzuki
Viruses.2022; 14(8): 1722. CrossRef - Molecular characteristics of a novel hypovirus from Trichoderma harzianum
Jeesun Chun, Kum-Kang So, Yo-Han Ko, Dae-Hyuk Kim
Archives of Virology.2022; 167(1): 233. CrossRef - Sustainable Management of Medicago sativa for Future Climates: Insect Pests, Endophytes and Multitrophic Interactions in a Complex Environment
Mark R. McNeill, Xiongbing Tu, Eric Altermann, Wu Beilei, Shengjing Shi
Frontiers in Agronomy.2022;[Epub] CrossRef - A New Double-Stranded RNA Mycovirus in Cryphonectria naterciae Is Able to Cross the Species Barrier and Is Deleterious to a New Host
Carolina Cornejo, Sakae Hisano, Helena Bragança, Nobuhiro Suzuki, Daniel Rigling
Journal of Fungi.2021; 7(10): 861. CrossRef
- Co-infection with two novel mycoviruses affects the biocontrol activity of Trichoderma polysporum
- Limiting the pathogenesis of Salmonella Typhimurium with berry phenolic extracts and linoleic acid overproducing Lactobacillus casei
- Zajeba Tabashsum , Mengfei Peng , Cassendra Bernhardt , Puja Patel , Michael Carrion , Shaik O. Rahaman , Debabrata Biswas
- J. Microbiol. 2020;58(6):489-498. Published online April 22, 2020
- DOI: https://doi.org/10.1007/s12275-020-9545-1
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Abstract
- The growing threat of emergent multidrug-resistant enteric bacterial pathogens, and their adopted virulence properties are directing to find alternative antimicrobials and/or development of dietaries that can improve host gut health and/or defense. Recently, we found that modified Lactobacillus casei (Lc + CLA) with increased production of conjugated linoleic acid has antimicrobial and other beneficial properties. Further, prebiotic alike products such as berry pomace extracts (BPEs), increase the growth of probiotics and inhibit the growth of certain bacterial pathogens. In this study, we evaluated the antibacterial effect of genetically modified Lc + CLA along with BPEs against major enteric pathogen Salmonella enterica serovar Typhimurium (ST). In mixed culture condition, the growth of ST was significantly reduced in the presence of Lc + CLA and/or BPEs. Bacterial cell-free cultural supernatant (CFCS) collected from wild-type Lc or modified Lc + CLA strains also inhibited the growth and survival of ST, and those inhibitory effects were enhanced in the presence of BPEs. We also found that the interaction of the pathogen with cultured host (HD-11 and INT-407) cells were also altered in the presence of either Lc or Lc + CLA strain or their CFCSs significantly. Furthermore, the relative expression of genes related to ST virulence and physicochemical properties of ST was altered by the effect of CFCSs of either Lc or Lc + CLA. These findings indicate that a diet containing synbiotic, specifically linoleic acid, over-produced Lc + CLA and prebiotic product BPEs, might have the potential to be effective in controlling ST growth and pathogenesis.
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Citations
Citations to this article as recorded by-
Natural anti-adhesive components against pathogenic bacterial adhesion and infection in gastrointestinal tract: case studies of
Helicobacter pylori
,
Salmonella enterica
,
Clostridiu
Xiaoyu Bao, Jianping Wu
Critical Reviews in Food Science and Nutrition.2024; : 1. CrossRef - Combined effect of metabolites produced by a modified Lactobacillus casei and berry phenolic extract on Campylobacter and microbiome in chicken cecum contents
Zajeba Tabashsum, Zabdiel Alvarado‐Martinez, Matthew J. Wall, Arpita Aditya, Debabrata Biswas
Journal of Food Science.2023; 88(6): 2583. CrossRef - Intracellular autolytic whole cell Salmonella vaccine prevents colonization of pathogenic Salmonella Typhimurium in chicken
Mengfei Peng, Jungsoo Joo, Zabdiel Alvarado-Martinez, Zajeba Tabashsum, Arpita Aditya, Debabrata Biswas
Vaccine.2022; 40(47): 6880. CrossRef - Lactobacilli, a Weapon to Counteract Pathogens through the Inhibition of Their Virulence Factors
Andrea Colautti, Elisabetta Orecchia, Giuseppe Comi, Lucilla Iacumin, Laurie E. Comstock
Journal of Bacteriology.2022;[Epub] CrossRef -
Florfenicol Enhances Colonization of a Salmonella enterica Serovar Enteritidis
floR
Mutant with Major Alterations to the Intestinal Microbiota and Metabolome in Neonatal Chickens
Xueran Mei, Boheng Ma, Xiwen Zhai, Anyun Zhang, Changwei Lei, Lei Zuo, Xin Yang, Changyu Zhou, Hongning Wang, Johanna Björkroth
Applied and Environmental Microbiology.2021;[Epub] CrossRef
-
Natural anti-adhesive components against pathogenic bacterial adhesion and infection in gastrointestinal tract: case studies of
Helicobacter pylori
,
Salmonella enterica
,
Clostridiu
- Construction of a genetically modified T7Select phage system to express the antimicrobial peptide 1018
- David J. Lemon , Matthew K. Kay , James K. Titus , April A. Ford , Wen Chen , LCDR Nicholas J. Hamlin , Yoon Y. Hwang
- J. Microbiol. 2019;57(6):532-538. Published online May 27, 2019
- DOI: https://doi.org/10.1007/s12275-019-8686-6
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- 23 Web of Science
- 23 Crossref
-
Abstract
-
Bacteriophage therapy was an ascendant technology for combating
bacterial infections before the golden age of antibiotics,
but the therapeutic potential of phages was largely ignored
after the discovery of penicillin. Recently, with antibioticresistant
infections on the rise, these phages are receiving renewed
attention to combat problematic bacterial infections.
Our approach is to enhance bacteriophages with antimicrobial
peptides, short peptides with broad-spectrum antibiotic or
antibiofilm effects. We inserted coding sequences for 1018,
an antimicrobial peptide previously shown to be an effective
broad-spectrum antimicrobial and antibiofilm agent, or the
fluorescent marker mCherry, into the T7Select phage genome.
Transcription and production of 1018 or mCherry began
rapidly after E. coli cultures were infected with genetically modified
phages. mCherry fluorescence, which requires a 90 min
initial maturation period, was observed in infected cultures
after 2 h of infection. Finally, we tested phages expressing 1018
(1018 T7) against bacterial planktonic cultures and biofilms,
and found the 1018 T7 phage was more effective than the
unmodified T7Select phage at both killing planktonic cells and
eradicating established biofilms, validating our phage-driven
antimicrobial peptide expression system. The combination
of narrow-spectrum phages delivering relatively high local
doses of broad-spectrum antimicrobials could be a powerful
method
to combat resistant infections. The experiments we describe prove this combination is feasible in vitro, but further testing and optimization are required before genetically modified phages are ready for use in vivo. -
Citations
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Xianxun Sun, Tao Tian, Yindong Lian, Zongqiang Cui
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International Journal of Clinical Practice.2024; 2024: 1. CrossRef - Designing a simple and efficient phage biocontainment system using the amber suppressor initiator tRNA
Pamela R. Tsoumbris, Russel M. Vincent, Paul R. Jaschke
Archives of Virology.2024;[Epub] CrossRef - Applications of designer phage encoding recombinant gene payloads
Daniel S. Schmitt, Sara D. Siegel, Kurt Selle
Trends in Biotechnology.2024; 42(3): 326. CrossRef - Identification and characterization of TatD DNase in planarian Dugesia japonica and its antibiofilm effect
Tong Yu, Zhe Sun, Xiangyu Cao, Fengtang Yang, Qiuxiang Pang, Hongkuan Deng
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Materials Science and Engineering: R: Reports.2023; 153: 100715. CrossRef - Genetic Engineering and Biosynthesis Technology: Keys to Unlocking the Chains of Phage Therapy
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Viruses.2023; 15(8): 1736. CrossRef - Engineering therapeutic phages for enhanced antibacterial efficacy
Susanne Meile, Jiemin Du, Matthew Dunne, Samuel Kilcher, Martin J Loessner
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Journal of Microbiology.2022; 60(1): 128. CrossRef - How Good are Bacteriophages as an Alternative Therapy to Mitigate Biofilms of Nosocomial Infections
Aditi Singh, Sudhakar Padmesh, Manish Dwivedi, Irena Kostova
Infection and Drug Resistance.2022; Volume 15: 503. CrossRef - Comparative Analysis of NanoLuc Luciferase and Alkaline Phosphatase Luminescence Reporter Systems for Phage-Based Detection of Bacteria
Shalini Wijeratne, Arindam Bakshi, Joey Talbert
Bioengineering.2022; 9(9): 479. CrossRef - Construction and Characterization of T7 Bacteriophages Harboring Apidaecin-Derived Sequences
Tobias Ludwig, Ralf Hoffmann, Andor Krizsan
Current Issues in Molecular Biology.2022; 44(6): 2554. CrossRef - Genetic and Chemical Engineering of Phages for Controlling Multidrug-Resistant Bacteria
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Robert E. W. Hancock, Morgan A. Alford, Evan F. Haney
Nature Reviews Microbiology.2021; 19(12): 786. CrossRef - Bacteriophage manipulation of the microbiome associated with tumour microenvironments-can this improve cancer therapeutic response?
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宇波 向
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Celia Ferriol-González, Pilar Domingo-Calap
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Bionatura.2020; 5(1): 1078. CrossRef - The Principles, Mechanisms, and Benefits of Unconventional Agents in the Treatment of Biofilm Infection
Jasminka Talapko, Ivana Škrlec
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- The antimicrobial potential of a new derivative of cathelicidin from Bungarus fasciatus against methicillin-resistant Staphylococcus aureus
- Mercedeh Tajbakhsh , Abdollah Karimi , Abolghasem Tohidpour , Naser Abbasi , Fatemeh Fallah , Maziar Mohammad Akhavan
- J. Microbiol. 2018;56(2):128-137. Published online February 2, 2018
- DOI: https://doi.org/10.1007/s12275-018-7444-5
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-
Abstract
- Cathelicidins are a family of antimicrobial peptides which exhibit broad antimicrobial activities against antibiotic-resistant bacteria. Considering the progressive antibiotic resistance, cathelicidin is a candidate for use as an alternative approach to treat and overcome the challenge of antimicrobial resistance. Cathelicidin-BF (Cath-BF) is a short antimicrobial peptide, which was originally extracted from the venom of Bungarus fasciatus. Recent studies have reported that Cath-BF and some related derivatives exert strong antimicrobial and weak hemolytic properties. This study investigates the bactericidal and cytotoxic effects of Cath-BF and its analogs (Cath-A and Cath-B). Cath-A and Cath-B were designed to increase their net positive charge, to have more activity against methicillin resistant S. aureus (MRSA). The results of this study show that Cath-A, with a +17-net charge, has the most noteworthy antimicrobial activity against MRSA strains, with minimum inhibitory concentration (MIC) ranging between 32–128 μg/ml. The bacterial kinetic analysis by 1 × MIC concentration of each peptide shows that Cath-A neutralizes the clinical MRSA isolate for 60 min. The present data support the notion that increasing the positive net charge of antimicrobial peptides can increase their potential antimicrobial activity. Cath-A also displayed the weakest cytotoxicity effect against human umbilical vein endothelial and H9c2 rat cardiomyoblast cell lines. Analysis of the hemolytic activity reveals that all three peptides exhibit minor hemolytic activity against human erythrocytes at concentrations up to 250 μg/ml. Altogether, these results suggest that Cath-A and Cath-B are competent candidates as novel antimicrobial compounds against MRSA and possibly other multidrug resistant bacteria.
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Citations
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Biomolecules.2018; 8(4): 118. CrossRef
- Synthetic peptide (DP1) functionalized graphene oxide: A biocompatible nanoformulation with broad-spectrum antibacterial and antibiofilm activity
- Phenotypic and genotypic correlates of daptomycin-resistant methicillin-susceptible Staphylococcus aureus clinical isolates
- Kyoung-Mi Kang , Nagendra N. Mishra , Kun Taek Park , Gi-Yong Lee , Yong Ho Park , Arnold S. Bayer , Soo-Jin Yang
- J. Microbiol. 2017;55(2):153-159. Published online January 26, 2017
- DOI: https://doi.org/10.1007/s12275-017-6509-1
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Abstract
- Daptomycin (DAP) has potent activity in vitro and in vivo against both methicillin-susceptible Staphylococcus aureus (MSSA) and methicillin-resistant S. aureus (MRSA) strains. DAP-resistance (DAP-R) in S. aureus has been mainly observed in MRSA strains, and has been linked to single nucleotide polymorphisms (SNPs) within the mprF gene leading to altered cell membrane (CM) phospholipid (PL) profiles, enhanced positive surface charge, and changes in CM fluidity. The current study was designed to delineate whether these same genotypic and phenotypic perturbations are demonstrated in clinically-derived DAP-R MSSA strains. We used three isogenic DAP-susceptible (DAP-S)/DAP-R strainpairs and compared: (i) presence of mprF SNPs, (ii) temporal expression profiles of the two key determinants (mprF and dltABCD) of net positive surface charge, (iii) increased production of mprF-dependent lysinylated-phosphatidylglycerol (L-PG), (iv) positive surface charge assays, and (v) susceptibility to cationic host defense peptides (HDPs) of neutrophil and platelet origins. Similar to prior data in MRSA, DAP-R (vs DAP-S) MSSA strains exhibited hallmark hot-spot SNPs in mprF, enhanced and dysregulated expression of both mprF and dltA, L-PG overproduction, HDP resistance and enhanced positive surface charge profiles. However, in contrast to most DAP-R MRSA strains, there were no changes in CM fluidity seen. Thus, charge repulsion via mprF- and dlt-mediated enhancement of positive surface charge may be the main mechanism to explain DAP-R in MSSA strains.
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Review
- MINIREVIEW] The therapeutic applications of antimicrobial peptides (AMPs): a patent review
- Hee-Kyoung Kang , Cheolmin Kim , Chang Ho Seo , Yoonkyung Park
- J. Microbiol. 2017;55(1):1-12. Published online December 30, 2016
- DOI: https://doi.org/10.1007/s12275-017-6452-1
- 48 View
- 0 Download
- 270 Crossref
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Abstract
- Antimicrobial peptides (AMPs) are small molecules with a broad spectrum of antibiotic activities against bacteria, yeasts, fungi, and viruses and cytotoxic activity on cancer cells, in addition to anti-inflammatory and immunomodulatory activities. Therefore, AMPs have garnered interest as novel therapeutic agents. Because of the rapid increase in drug-resistant pathogenic microorganisms, AMPs from synthetic and natural sources have been developed using alternative antimicrobial strategies. This article presents a broad analysis of patents referring to the therapeutic applications of AMPs since 2009. The review focuses on the universal trends in the effective design, mechanism, and biological evolution of AMPs.
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Polymers.2020; 12(5): 1195. CrossRef - Effects of Peptide Thanatin on the Growth and Transcriptome of Penicillium digitatum
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Infection, Genetics and Evolution.2017; 54: 238. CrossRef - Testing cathelicidin susceptibility of bacterial mastitis isolates: Technical challenges and data output for clinical isolates
Melissa N. Langer, Stefanie Blodkamp, Martin Bayerbach, Andrea T. Feßler, Nicole de Buhr, Thomas Gutsmann, Lothar Kreienbrock, Stefan Schwarz, Maren von Köckritz-Blickwede
Veterinary Microbiology.2017; 210: 107. CrossRef - Protein-only, antimicrobial peptide-containing recombinant nanoparticles with inherent built-in antibacterial activity
Naroa Serna, Laura Sánchez-García, Alejandro Sánchez-Chardi, Ugutz Unzueta, Mónica Roldán, Ramón Mangues, Esther Vázquez, Antonio Villaverde
Acta Biomaterialia.2017; 60: 256. CrossRef
- Boc‐Protected Phenylalanine and Tryptophan‐Based Dipeptides: A Broad Spectrum Anti‐Bacterial Agent
Research Support, Non-U.S. Gov'ts
- The Intracellular Mechanism of Action on Escherichia coli of BF2-A/C, Two Analogues of the Antimicrobial Peptide Buforin 2
- Gang Hao , Yong-Hui Shi , Ya-Li Tang , Guo-Wei Le
- J. Microbiol. 2013;51(2):200-206. Published online April 27, 2013
- DOI: https://doi.org/10.1007/s12275-013-2441-1
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- 32 Scopus
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Abstract
- In the present study, the antimicrobial peptides BF2-A and BF2-C, two analogues of Buforin 2, were chemically synthesized and the activities were assayed. To elucidate the bactericidal mechanism of BF2-A/C and their different antimicrobial activities, the influence of peptides to E. coli cell membrane and targets of intracellular action were researched. Obviously, BF2-A and BF2-C did not induce the influx of PI into the E. coli cells, indicating nonmemebrane permeabilizing killing action. The FITC-labeled BF2-A/C could penetrate the E. coli cell membrane and BF2-C penetrated the cells more efficiently. Furthermore, BF2-A/C could bind to DNA and RNA respectively, and the affinity of BF2-C to DNA was powerful at least over 4 times than that of BF2-A. The present results implied that BF2-A and BF2-C inhibited the cellular functions by binding to DNA and RNA of cells after penetrating the cell membranes, resulting in the rapid cell death. The structure-activity relationship analysis of BF2-A/C revealed that the cell-penetrating efficiency and the affinity ability to DNA were critical factors for determining the antimicrobial potency of both peptides. The more efficient cellpenetrating and stronger affinity to DNA caused that BF2-C displayed more excellent antimicrobial activity and rapid killing kinetics than BF2-A.
- Expression of Recombinant Hybrid Peptide Hinnavin II/α-Melanocyte-Stimulating Hormone in Escherichia coli: Purification and Characterization
- Son Kwon Bang , Chang Soo Kang , Man-Deuk Han , In Seok Bang
- J. Microbiol. 2010;48(1):24-29. Published online March 11, 2010
- DOI: https://doi.org/10.1007/s12275-009-0317-1
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- 7 Scopus
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Abstract
- The increasing problem of antibiotic resistance among pathogenic bacteria requires novel strategies for the construction of multiple, joined genes of antimicrobial agents. The strategy used in this study involved synthesis of a cDNA-encoding hinnavin II/α-melanocyte-stimulating hormone (hin/MSH) hybrid peptide, which was cloned into the pET32a (+) vector to allow expression of the hybrid peptide as a fusion protein in Escherichia coli BL21 (DE3). The resulting expression of fusion protein Trx-hin/MSH could reach up to 20% of the total cell proteins. More than 50% of the target protein was in a soluble form. The target fusion protein from the soluble fraction, Trx-hin/MSH, was easily purified by Ni2+-chelating chromatography. Then, enterokinase cleavage effectively cleaved the Trx-hin/MSH to release the combinant hin/MSH (rhin/MSH) hybrid peptide. After removing the contaminants, we purified the recombinant hybrid peptide to homogeneity by reversed-phase FPLC and obtained 210 mg of pure, active rhin/MSH from 800 ml of culture medium. Antimicrobial activity assay demonstrated that rhin/MSH had a broader spectrum of activity than did the parental hinnavin II or MSH against fungi and Gram-positive and Gram-negative bacteria. These results suggest an efficient method for producing high-level expression of various kinds of antimicrobial peptides that are toxic to the host, a reliable and simple method for producing different hybrid peptides for biological studies.
- Note] Antibacterial Activity of Recombinant hCAP18/LL37 Protein Secreted from Pichia pastoris
- Soon-ja Kim , Renshu Quan , Sung-Jin Lee , Hak-Kyo Lee , Joong-Kook Choi
- J. Microbiol. 2009;47(3):358-362. Published online June 26, 2009
- DOI: https://doi.org/10.1007/s12275-009-0131-9
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- 17 Scopus
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Abstract
- Human antimicrobial peptide CAP18/LL37 (hCAP18/LL37) was expressed in Pichia pastoris and its antibacterial activity was tested against pathogenic bacteria. The full length ORF of hCAP18/LL37 was cloned into the pPICZαA vector followed by integration into the genomic AOX1 gene of P. pastoris. Agar diffusion assay demonstrated that the different hCAP18/LL37 transformants showed various antibacterial activities against Staphylococcus aureus, Micrococcus luteus, and Salmonella gastroenteritis. The secreted form of hCAP18/LL37 exhibited its maximum activity after 72 h incubation with 2% methanol in MM media, not in BMM. This result suggests that the yeast secreted expression system can be used as a production tool of antimicrobial peptides for industrial or pharmaceutical application.
Journal Article
- Biologically Active and C-Amidated HinnavinII-38-Asn Produced from a Trx Fusion Construct in Escherichia coli
- Chang Soo Kang , Seung-Yeol Son , In Seok Bang
- J. Microbiol. 2008;46(6):656-661. Published online December 24, 2008
- DOI: https://doi.org/10.1007/s12275-008-0214-z
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- 9 Scopus
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Abstract
- The cabbage butterfly (Artogeia rapae) antimicrobial peptide hinnavinII as a member of cecropin family is synthesized as 37 residues in size with an amidated lysine at C-terminus and shows the humoral immune response to a bacterial invasion. In this work, a synthetic gene for hinnavinII-38-Asn (HIN) with an additional amino acid asparagine residue containing amide group at C-terminus was cloned into pET-32a(+) vector to allow expression of HIN as a Trx fusion protein in Escherichia coli strain BL21 (DE3) pLysS. The resulting expression level of the fusion protein Trx-HIN could reach 15~20% of the total cell proteins and more than 70% of the target proteins were in soluble form. The fusion protein could be purified successfully by HiTrap Chelating HP column and a high yield of 15 mg purified fusion protein was obtained from 80 ml E. coli culture. Recombinant HIN was readily obtained by enterokinase cleavage of the fusion protein followed by FPLC chromatography, and 3.18 mg pure active recombinant HIN was obtained from 80 ml culture. The molecular mass of recombinant HIN determined by MALDI-TOF mass spectrometer is 4252.084 Da which matches the theoretical mass (4252.0 Da) of HIN. Comparing the antimicrobial activities of the recombinant hinnavinII with C-amidated terminus to that without an amidated C-terminus, we found that the amide of asparagine at C-terminus of hinnavinII improved its potency on certain microorganism such as E. coli, Enterobacter cloacae, Bacillus megaterium, and Staphylococcus aureus.
- Regulation of Class II Bacteriocin Production by Cell-Cell Signaling
- Luis E. N. Quadri
- J. Microbiol. 2003;41(3):175-182.
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Abstract
- Production of ribosomally synthesized antimicrobial peptides usually referred to as bacteriocins is an inducible trait in several gram positive bacteria, particularly in those belonging to the group of lactic acid bacteria. In many of these organisms, production of bacteriocins is inducible and induction requires secretion and extracellular accumulation of peptides that act as chemical messengers and trigger bacteriocin production. These inducer peptides are often referred to as autoinducers and are believed to permit a quorum sensing-based regulation of bacteriocin production. Notably, the peptides acting as autoinducers are dedicated peptides with or without antimicrobial activity or the bacteriocins themselves. The autoinducer-dependent induction of bacteriocin production requires histidine protein kinases and response regulator proteins of two-component signal transduction systems. The current working model for the regulation of class II bacteriocin production in lactic acid bacteria and the most relevant direct and indirect pieces of evidence supporting the model are discussed in this minireview.
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