β-Lytic Protease of Lysobacter capsici VKM B-2533T

Abstract

Bacteriolytic enzymes are promising antimicrobial agents for developing new-generation drugs. Recently, we have isolated a β-lytic protease (BlpLc) from the culture liquid of Lysobacter capsici VKM B-2533T. This BlpLc possesses a valuable property, not described for β-lytic proteases (Blps) earlier, of hydrolyzing living cells of Staphylococcus aureus 55 MRSA clinical isolate. This work phylogenetically characterized the BlpLc and investigated its properties. Analysis revealed a variability of pre-/pro-parts of Blp precursors. The mature BlpLc is the closest to the earlier annotated but not isolated Blp from Lysobacter sp. Root690. The biochemical characterization found conditions for the BlpLc general bacteriolytic activity relative to autoclaved S. aureus 209P cells to differ from that of earlier isolated Blp. Unexpected was the effect of serine (phenylmethylsulfonyl fluoride (PMSF)) and cysteine (p-chloromercuribenzoate (p-CMB)) protease inhibitors on BlpLc bacteriolytic and proteolytic activities. The specificity of BlpLc proteolytic action relative to hemoglobin, elastin, gelatin, collagen, azofibrin, myoglobin, ovalbumin, and ovamucoid was found. New types of peptide bonds—Gly-X, Ser-X, Lys-X, Ala-X, Val-X, Glu-X, and Phe-X—hydrolyzed by the enzyme in protein substrates were first revealed using MALDI-TOF. Turbidimetrically, the BlpLc was found to lyze living cells of S. aureus 209P, Micrococcus luteus B1819, and M. roseus B1236, which is important for expanding the enzyme’s applied properties.

Whole-Genome Sequencing of Lysobacter capsici VKM B-2533 T and Lysobacter gummosus 10.1.1, Promising Producers of Lytic Agents

“…Some bacteria of the genus Lysobacter are capable of producing various antimicrobial agents, such as antibiotics, bacteriolytic enzymes, and peptides ( 1 – 7 ). Despite their great promise for biomedicine, these bacteria have been studied very poorly.…”

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

“…IB−9374, and L. capsici VKM B−2533 T [3][4][5]. The isolated enzymes have been characterized to various degrees [3,4,6]. In the peptidoglycan of target cells, the enzyme digests the bonds in the interpeptide bridge [4].…”

“…In the peptidoglycan of target cells, the enzyme digests the bonds in the interpeptide bridge [4]. Recently, we have isolated a Blp from the culture fluid of L. capsici VKM B−2533 T and characterized it [5,6]. Apart from hydrolysing substrates for proteases, the enzyme is highly efficient in digesting the peptidoglycan of living Micrococcus luteus Ac−2230 T and Staphylococcus aureus 55 MRSA cells.…”

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

“…To date, however, only lysozyme has been widely used in medicine, agriculture, and the food industry. At the same time, many bacteriolytic enzymes, Blp included, are not inferior to, and even surpass the lytic action of lysozyme, but are not used widely in practice [ 4 , 5 , 6 ]. This is primarily due to the difficulties of obtaining effective expression systems for such enzymes, which include the “toxicity” of target proteins for producer cells, the absence of secretory pathways in recombinant strains that may be necessary for the correct folding of target proteins, and refolding problems in obtaining proteins from inclusion bodies [ 7 , 8 , 9 , 10 ].…”

“…XL1 [ 11 , 12 , 13 , 14 , 15 , 16 , 17 ]. Our recent research has been related to the study of antimicrobial potential in L. capsici VKM B-2533 T [ 6 , 10 ]. Thus, it has been found that this bacterium has a potent antimicrobial effect against bacteria, fungi, and yeasts.…”