Cargando…
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...
Autores principales: | , , , |
---|---|
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 |
_version_ | 1784716352524648448 |
---|---|
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. |
format | Online Article Text |
id | pubmed-9145596 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
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 |
work_keys_str_mv | AT kudryakovairina thefirsthomologousexpressionsystemfortheblyticproteaseoflysobactercapsicivkmb2533tapromisingantimicrobialagent AT afoshinalexey thefirsthomologousexpressionsystemfortheblyticproteaseoflysobactercapsicivkmb2533tapromisingantimicrobialagent AT leontyevskayaelena thefirsthomologousexpressionsystemfortheblyticproteaseoflysobactercapsicivkmb2533tapromisingantimicrobialagent AT leontyevskayavasilyevanatalia thefirsthomologousexpressionsystemfortheblyticproteaseoflysobactercapsicivkmb2533tapromisingantimicrobialagent AT kudryakovairina firsthomologousexpressionsystemfortheblyticproteaseoflysobactercapsicivkmb2533tapromisingantimicrobialagent AT afoshinalexey firsthomologousexpressionsystemfortheblyticproteaseoflysobactercapsicivkmb2533tapromisingantimicrobialagent AT leontyevskayaelena firsthomologousexpressionsystemfortheblyticproteaseoflysobactercapsicivkmb2533tapromisingantimicrobialagent AT leontyevskayavasilyevanatalia firsthomologousexpressionsystemfortheblyticproteaseoflysobactercapsicivkmb2533tapromisingantimicrobialagent |