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

A successful homologous expression system based on VKM B-2533 and the plasmid pBBR1-MCS5 was first developed for a promising bacteriolytic enzyme of this bacterium, β-lytic protease (Blp). In the expression strains, 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 P- and P-, 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 P- 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 VKM B-2533. The expression of the gene in P- increased by 585-fold. The developed homologous system for the gene expression of bacteriolytic enzymes is potentially biotechnologically valuable, and is promising for creating highly efficient expression strains.