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Engineering and Purification of Microcin C7 Variants Resistant to Trypsin and Analysis of Their Biological Activity

Microcin C7 (McC) as a viable form of antimicrobial has gained substantial attention due to its distinctive antimicrobial activity, by targeting aspartyl tRNA synthetase. McC can be a potential solution against pathogenic microbial infections in the postantibiotic era. However, considering that degr...

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Autores principales: Yang, Guangxin, Shang, Lijun, Liu, Lu, Li, Zeqiang, Zeng, Xiangfang, Ding, Xiuliang, Huang, Jinxiu, Qiao, Shiyan, Yu, Haitao
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10525924/
https://www.ncbi.nlm.nih.gov/pubmed/37760643
http://dx.doi.org/10.3390/antibiotics12091346
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author Yang, Guangxin
Shang, Lijun
Liu, Lu
Li, Zeqiang
Zeng, Xiangfang
Ding, Xiuliang
Huang, Jinxiu
Qiao, Shiyan
Yu, Haitao
author_facet Yang, Guangxin
Shang, Lijun
Liu, Lu
Li, Zeqiang
Zeng, Xiangfang
Ding, Xiuliang
Huang, Jinxiu
Qiao, Shiyan
Yu, Haitao
author_sort Yang, Guangxin
collection PubMed
description Microcin C7 (McC) as a viable form of antimicrobial has gained substantial attention due to its distinctive antimicrobial activity, by targeting aspartyl tRNA synthetase. McC can be a potential solution against pathogenic microbial infections in the postantibiotic era. However, considering that degradation by digestive enzymes can disrupt the function of this peptide in the gastrointestinal tract, in this study, we attempt to design McC variants to overcome several barriers that may affect its stability and biological activity. The mccA gene encoding the McC peptide precursor was mutated and 12 new McC variants with trypsin resistance were found. The Yej(+)rimL(−) strain was used as an indicator to determine the minimum inhibitory concentrations (MICs). The results showed that three variants, including R2A, R2T and R2Q, among 12 variants formed by the replacement of the second arginine of the McC peptide with different amino acids, were resistant to trypsin and had an outstanding antimicrobial ability, with MIC values of 12.5, 25, and 25 μg/mL, respectively. Taken together, our findings show that the engineering of the site-directed mutagenesis of McC significantly enhances McC trypsin resistance and maintains a great antimicrobial activity.
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spelling pubmed-105259242023-09-28 Engineering and Purification of Microcin C7 Variants Resistant to Trypsin and Analysis of Their Biological Activity Yang, Guangxin Shang, Lijun Liu, Lu Li, Zeqiang Zeng, Xiangfang Ding, Xiuliang Huang, Jinxiu Qiao, Shiyan Yu, Haitao Antibiotics (Basel) Article Microcin C7 (McC) as a viable form of antimicrobial has gained substantial attention due to its distinctive antimicrobial activity, by targeting aspartyl tRNA synthetase. McC can be a potential solution against pathogenic microbial infections in the postantibiotic era. However, considering that degradation by digestive enzymes can disrupt the function of this peptide in the gastrointestinal tract, in this study, we attempt to design McC variants to overcome several barriers that may affect its stability and biological activity. The mccA gene encoding the McC peptide precursor was mutated and 12 new McC variants with trypsin resistance were found. The Yej(+)rimL(−) strain was used as an indicator to determine the minimum inhibitory concentrations (MICs). The results showed that three variants, including R2A, R2T and R2Q, among 12 variants formed by the replacement of the second arginine of the McC peptide with different amino acids, were resistant to trypsin and had an outstanding antimicrobial ability, with MIC values of 12.5, 25, and 25 μg/mL, respectively. Taken together, our findings show that the engineering of the site-directed mutagenesis of McC significantly enhances McC trypsin resistance and maintains a great antimicrobial activity. MDPI 2023-08-22 /pmc/articles/PMC10525924/ /pubmed/37760643 http://dx.doi.org/10.3390/antibiotics12091346 Text en © 2023 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Yang, Guangxin
Shang, Lijun
Liu, Lu
Li, Zeqiang
Zeng, Xiangfang
Ding, Xiuliang
Huang, Jinxiu
Qiao, Shiyan
Yu, Haitao
Engineering and Purification of Microcin C7 Variants Resistant to Trypsin and Analysis of Their Biological Activity
title Engineering and Purification of Microcin C7 Variants Resistant to Trypsin and Analysis of Their Biological Activity
title_full Engineering and Purification of Microcin C7 Variants Resistant to Trypsin and Analysis of Their Biological Activity
title_fullStr Engineering and Purification of Microcin C7 Variants Resistant to Trypsin and Analysis of Their Biological Activity
title_full_unstemmed Engineering and Purification of Microcin C7 Variants Resistant to Trypsin and Analysis of Their Biological Activity
title_short Engineering and Purification of Microcin C7 Variants Resistant to Trypsin and Analysis of Their Biological Activity
title_sort engineering and purification of microcin c7 variants resistant to trypsin and analysis of their biological activity
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10525924/
https://www.ncbi.nlm.nih.gov/pubmed/37760643
http://dx.doi.org/10.3390/antibiotics12091346
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