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β-Lytic Protease of Lysobacter capsici VKM B-2533(T)
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-2533(T). This BlpLc possesses a valuable property, not described for β-lytic proteases (Blps) earlie...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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MDPI
2020
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7693508/ https://www.ncbi.nlm.nih.gov/pubmed/33126447 http://dx.doi.org/10.3390/antibiotics9110744 |
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author | Afoshin, Alexey S. Konstantinov, Mihail A. Toropygin, Ilya Yu. Kudryakova, Irina V. Vasilyeva, Natalia V. |
author_facet | Afoshin, Alexey S. Konstantinov, Mihail A. Toropygin, Ilya Yu. Kudryakova, Irina V. Vasilyeva, Natalia V. |
author_sort | Afoshin, Alexey S. |
collection | PubMed |
description | 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-2533(T). 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. |
format | Online Article Text |
id | pubmed-7693508 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-76935082020-11-28 β-Lytic Protease of Lysobacter capsici VKM B-2533(T) Afoshin, Alexey S. Konstantinov, Mihail A. Toropygin, Ilya Yu. Kudryakova, Irina V. Vasilyeva, Natalia V. Antibiotics (Basel) Article 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-2533(T). 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. MDPI 2020-10-28 /pmc/articles/PMC7693508/ /pubmed/33126447 http://dx.doi.org/10.3390/antibiotics9110744 Text en © 2020 by the authors. 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 (http://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Afoshin, Alexey S. Konstantinov, Mihail A. Toropygin, Ilya Yu. Kudryakova, Irina V. Vasilyeva, Natalia V. β-Lytic Protease of Lysobacter capsici VKM B-2533(T) |
title | β-Lytic Protease of Lysobacter capsici VKM B-2533(T) |
title_full | β-Lytic Protease of Lysobacter capsici VKM B-2533(T) |
title_fullStr | β-Lytic Protease of Lysobacter capsici VKM B-2533(T) |
title_full_unstemmed | β-Lytic Protease of Lysobacter capsici VKM B-2533(T) |
title_short | β-Lytic Protease of Lysobacter capsici VKM B-2533(T) |
title_sort | β-lytic protease of lysobacter capsici vkm b-2533(t) |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7693508/ https://www.ncbi.nlm.nih.gov/pubmed/33126447 http://dx.doi.org/10.3390/antibiotics9110744 |
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