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

Abstract

A successful homologous expression system based on Lysobacter capsici VKM B-2533T 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 PGroEL(A)-blp and L. capsici PT5-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 PT5-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-2533T. The expression of the serp gene in L. capsici PT5-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.

Structural and Functional Characterization of β−lytic Protease from Lysobacter capsici VKM B−2533T

“…We have recently isolated a Blp from the culture fluid of L. capsici VKM B−2533 T and characterized it [5]. An efficient homologous expression system for this enzyme was developed [30]. The present work aimed to establish the spatial structure of Blp.…”

Transcriptomic Analysis Followed by the Isolation of Extracellular Bacteriolytic Proteases from Lysobacter capsici VKM B-2533T

“…We have isolated αand β-lytic proteases, lysine-specific proteases, as well as identified new bacteriolytic proteases of MW 26 and 29 kDa, including N-acetylglucosaminidase [6]. For the β-lytic protease, the spatial structure has been established and its characterization has been expanded; an effective expression system has been developed [7,8]. The development of this system will enable us to isolate and study bacteriolytic Lysobacter enzymes with high efficiency.…”