The First Homologous Expression System for the β-Lytic Protease of Lysobacter capsici VKM B-2533T,a Promising Antimicrobial Agent |
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| Authors: | Irina Kudryakova Alexey Afoshin Elena Leontyevskaya Natalia Leontyevskaya |
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| Affiliation: | Laboratory of Microbial Cell Surface Biochemistry, G. K. Skryabin Institute of Biochemistry and Physiology of Microorganisms, Russian Academy of Sciences, Pushchino Center for Biological Research, 5 Prosp. Nauki, 142290 Pushchino, Russia; (I.K.); (A.A.); (E.L.) |
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| 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. |
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| Keywords: | homologous expression system Lysobacter staphylolytic β -lytic protease GroEL(A) and T5 promoters bacteriolytic enzymes |
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