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The First Homologous Expression System for the β-Lytic Protease of Lysobacter capsici VKM B-2533(T), a Promising Antimicrobial Agent

A successful homologous expression system based on Lysobacter capsici VKM B-2533(T) and the plasmid pBBR1-MCS5 was first developed for a promising bacteriolytic enzyme of this bacterium, β-lytic protease (Blp). In the expression strains, blp gene expression under the regulation of the GroEL(A) and T...

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Autores principales: Kudryakova, Irina, Afoshin, Alexey, Leontyevskaya, Elena, Leontyevskaya (Vasilyeva), Natalia
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9145596/
https://www.ncbi.nlm.nih.gov/pubmed/35628535
http://dx.doi.org/10.3390/ijms23105722
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author Kudryakova, Irina
Afoshin, Alexey
Leontyevskaya, Elena
Leontyevskaya (Vasilyeva), Natalia
author_facet Kudryakova, Irina
Afoshin, Alexey
Leontyevskaya, Elena
Leontyevskaya (Vasilyeva), Natalia
author_sort Kudryakova, Irina
collection PubMed
description A successful homologous expression system based on Lysobacter capsici VKM B-2533(T) and the plasmid pBBR1-MCS5 was first developed for a promising bacteriolytic enzyme of this bacterium, β-lytic protease (Blp). In the expression strains, blp gene expression under the regulation of the GroEL(A) and T5 promoters increased by 247- and 667-fold, respectively, as compared with the wild-type strain. After the cultivation of the expression strains L. capsici P(GroEL(A))-blp and L. capsici P(T5)-blp, the Blp yield increased by 6.7- and 8.5-fold, respectively, with respect to the wild-type strain. The cultivation of the expression strain L. capsici P(T5)-blp was successfully scaled up. Under fermentation conditions the yield of the enzyme increased by 1.6-fold. The developed homologous system was used to express the gene of the bacteriolytic serine protease (Serp) of L. capsici VKM B-2533(T). The expression of the serp gene in L. capsici P(T5)-serp increased by 585-fold. The developed homologous system for the gene expression of bacteriolytic Lysobacter enzymes is potentially biotechnologically valuable, and is promising for creating highly efficient expression strains.
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spelling pubmed-91455962022-05-29 The First Homologous Expression System for the β-Lytic Protease of Lysobacter capsici VKM B-2533(T), a Promising Antimicrobial Agent Kudryakova, Irina Afoshin, Alexey Leontyevskaya, Elena Leontyevskaya (Vasilyeva), Natalia Int J Mol Sci Article A successful homologous expression system based on Lysobacter capsici VKM B-2533(T) and the plasmid pBBR1-MCS5 was first developed for a promising bacteriolytic enzyme of this bacterium, β-lytic protease (Blp). In the expression strains, blp gene expression under the regulation of the GroEL(A) and T5 promoters increased by 247- and 667-fold, respectively, as compared with the wild-type strain. After the cultivation of the expression strains L. capsici P(GroEL(A))-blp and L. capsici P(T5)-blp, the Blp yield increased by 6.7- and 8.5-fold, respectively, with respect to the wild-type strain. The cultivation of the expression strain L. capsici P(T5)-blp was successfully scaled up. Under fermentation conditions the yield of the enzyme increased by 1.6-fold. The developed homologous system was used to express the gene of the bacteriolytic serine protease (Serp) of L. capsici VKM B-2533(T). The expression of the serp gene in L. capsici P(T5)-serp increased by 585-fold. The developed homologous system for the gene expression of bacteriolytic Lysobacter enzymes is potentially biotechnologically valuable, and is promising for creating highly efficient expression strains. MDPI 2022-05-20 /pmc/articles/PMC9145596/ /pubmed/35628535 http://dx.doi.org/10.3390/ijms23105722 Text en © 2022 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
Kudryakova, Irina
Afoshin, Alexey
Leontyevskaya, Elena
Leontyevskaya (Vasilyeva), Natalia
The First Homologous Expression System for the β-Lytic Protease of Lysobacter capsici VKM B-2533(T), a Promising Antimicrobial Agent
title The First Homologous Expression System for the β-Lytic Protease of Lysobacter capsici VKM B-2533(T), a Promising Antimicrobial Agent
title_full The First Homologous Expression System for the β-Lytic Protease of Lysobacter capsici VKM B-2533(T), a Promising Antimicrobial Agent
title_fullStr The First Homologous Expression System for the β-Lytic Protease of Lysobacter capsici VKM B-2533(T), a Promising Antimicrobial Agent
title_full_unstemmed The First Homologous Expression System for the β-Lytic Protease of Lysobacter capsici VKM B-2533(T), a Promising Antimicrobial Agent
title_short The First Homologous Expression System for the β-Lytic Protease of Lysobacter capsici VKM B-2533(T), a Promising Antimicrobial Agent
title_sort first homologous expression system for the β-lytic protease of lysobacter capsici vkm b-2533(t), a promising antimicrobial agent
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9145596/
https://www.ncbi.nlm.nih.gov/pubmed/35628535
http://dx.doi.org/10.3390/ijms23105722
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