Cargando…
Biochemical Characterization of An Arginine-Specific Alkaline Trypsin from Bacillus licheniformis
In the present study, we isolated a trypsin-producing strain DMN6 from the leather waste and identified it as Bacillus licheniformis through a two-step screening strategy. The trypsin activity was increased up to 140 from 20 U/mL through culture optimization. The enzyme was purified to electrophoret...
Autores principales: | , , , , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2015
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4691143/ https://www.ncbi.nlm.nih.gov/pubmed/26694369 http://dx.doi.org/10.3390/ijms161226200 |
_version_ | 1782407109854363648 |
---|---|
author | Gong, Jin-Song Li, Wei Zhang, Dan-Dan Xie, Min-Feng Yang, Biao Zhang, Rong-Xian Li, Heng Lu, Zhen-Ming Xu, Zheng-Hong Shi, Jin-Song |
author_facet | Gong, Jin-Song Li, Wei Zhang, Dan-Dan Xie, Min-Feng Yang, Biao Zhang, Rong-Xian Li, Heng Lu, Zhen-Ming Xu, Zheng-Hong Shi, Jin-Song |
author_sort | Gong, Jin-Song |
collection | PubMed |
description | In the present study, we isolated a trypsin-producing strain DMN6 from the leather waste and identified it as Bacillus licheniformis through a two-step screening strategy. The trypsin activity was increased up to 140 from 20 U/mL through culture optimization. The enzyme was purified to electrophoretic homogeneity with a molecular mass of 44 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and the specific activity of purified enzyme is 350 U/mg with Nα-Benzoyl-l-arginine ethylester as the substrate. The optimum temperature and pH for the trypsin are 65 °C and pH 9.0, respectively. Also, the enzyme can be significantly activated by Ba(2+). This enzyme is relatively stable in alkaline environment and displays excellent activity at low temperatures. It could retain over 95% of enzyme activity after 180 min of incubation at 45 °C. The distinguished activity under low temperature and prominent stability enhance its catalytic potential. In the current work, the open reading frame was obtained with a length of 1371 nucleotides that encoded a protein of 456 amino acids. These data would warrant the B. licheniformis trypsin as a promising candidate for catalytic application in collagen preparation and leather bating through further protein engineering. |
format | Online Article Text |
id | pubmed-4691143 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2015 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-46911432016-01-06 Biochemical Characterization of An Arginine-Specific Alkaline Trypsin from Bacillus licheniformis Gong, Jin-Song Li, Wei Zhang, Dan-Dan Xie, Min-Feng Yang, Biao Zhang, Rong-Xian Li, Heng Lu, Zhen-Ming Xu, Zheng-Hong Shi, Jin-Song Int J Mol Sci Article In the present study, we isolated a trypsin-producing strain DMN6 from the leather waste and identified it as Bacillus licheniformis through a two-step screening strategy. The trypsin activity was increased up to 140 from 20 U/mL through culture optimization. The enzyme was purified to electrophoretic homogeneity with a molecular mass of 44 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and the specific activity of purified enzyme is 350 U/mg with Nα-Benzoyl-l-arginine ethylester as the substrate. The optimum temperature and pH for the trypsin are 65 °C and pH 9.0, respectively. Also, the enzyme can be significantly activated by Ba(2+). This enzyme is relatively stable in alkaline environment and displays excellent activity at low temperatures. It could retain over 95% of enzyme activity after 180 min of incubation at 45 °C. The distinguished activity under low temperature and prominent stability enhance its catalytic potential. In the current work, the open reading frame was obtained with a length of 1371 nucleotides that encoded a protein of 456 amino acids. These data would warrant the B. licheniformis trypsin as a promising candidate for catalytic application in collagen preparation and leather bating through further protein engineering. MDPI 2015-12-17 /pmc/articles/PMC4691143/ /pubmed/26694369 http://dx.doi.org/10.3390/ijms161226200 Text en © 2015 by the authors; licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons by Attribution (CC-BY) license (http://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Gong, Jin-Song Li, Wei Zhang, Dan-Dan Xie, Min-Feng Yang, Biao Zhang, Rong-Xian Li, Heng Lu, Zhen-Ming Xu, Zheng-Hong Shi, Jin-Song Biochemical Characterization of An Arginine-Specific Alkaline Trypsin from Bacillus licheniformis |
title | Biochemical Characterization of An Arginine-Specific Alkaline Trypsin from Bacillus licheniformis |
title_full | Biochemical Characterization of An Arginine-Specific Alkaline Trypsin from Bacillus licheniformis |
title_fullStr | Biochemical Characterization of An Arginine-Specific Alkaline Trypsin from Bacillus licheniformis |
title_full_unstemmed | Biochemical Characterization of An Arginine-Specific Alkaline Trypsin from Bacillus licheniformis |
title_short | Biochemical Characterization of An Arginine-Specific Alkaline Trypsin from Bacillus licheniformis |
title_sort | biochemical characterization of an arginine-specific alkaline trypsin from bacillus licheniformis |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4691143/ https://www.ncbi.nlm.nih.gov/pubmed/26694369 http://dx.doi.org/10.3390/ijms161226200 |
work_keys_str_mv | AT gongjinsong biochemicalcharacterizationofanargininespecificalkalinetrypsinfrombacilluslicheniformis AT liwei biochemicalcharacterizationofanargininespecificalkalinetrypsinfrombacilluslicheniformis AT zhangdandan biochemicalcharacterizationofanargininespecificalkalinetrypsinfrombacilluslicheniformis AT xieminfeng biochemicalcharacterizationofanargininespecificalkalinetrypsinfrombacilluslicheniformis AT yangbiao biochemicalcharacterizationofanargininespecificalkalinetrypsinfrombacilluslicheniformis AT zhangrongxian biochemicalcharacterizationofanargininespecificalkalinetrypsinfrombacilluslicheniformis AT liheng biochemicalcharacterizationofanargininespecificalkalinetrypsinfrombacilluslicheniformis AT luzhenming biochemicalcharacterizationofanargininespecificalkalinetrypsinfrombacilluslicheniformis AT xuzhenghong biochemicalcharacterizationofanargininespecificalkalinetrypsinfrombacilluslicheniformis AT shijinsong biochemicalcharacterizationofanargininespecificalkalinetrypsinfrombacilluslicheniformis |