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Purification of Protease from Pseudomonas thermaerum GW1 Isolated from Poultry Waste Site
An extracellular protease was purified from Pseudomonas thermaerum GW1 a new strain identified by morphological, biochemical and 16S rDNA sequencing. It was isolated from soil of Poultry waste site at Ghazipur near Ghaziabad, Delhi. The strain produces extra cellular protease in the culture media th...
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Formato: | Texto |
Lenguaje: | English |
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Bentham Open
2010
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3048348/ https://www.ncbi.nlm.nih.gov/pubmed/21379398 http://dx.doi.org/10.2174/1874285801004010067 |
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author | Gaur, Smriti Agrahari, Sarita Wadhwa, Neeraj |
author_facet | Gaur, Smriti Agrahari, Sarita Wadhwa, Neeraj |
author_sort | Gaur, Smriti |
collection | PubMed |
description | An extracellular protease was purified from Pseudomonas thermaerum GW1 a new strain identified by morphological, biochemical and 16S rDNA sequencing. It was isolated from soil of Poultry waste site at Ghazipur near Ghaziabad, Delhi. The strain produces extra cellular protease in the culture media that was maintained at 37°C, 140 rpm. The media was harvested for protease after 48 hrs of incubation at 37°C in basal media supplemented with 1% casein. We report 6.08 fold purification of enzyme following ammonium sulphate precipitation and DEAE-cellulose chromatography. The molecular weight of the enzyme was estimated to be approximately 43,000 daltons as shown by casein zymography studies. The optimum pH for the proteolytic activity was pH 8.0 and enzyme remained stable between pH 5 -11 at 60°C. Interestingly Mn(2+) (5mM) activated enzyme activity by 5 fold, while Cu(2+), Mg(2+)and Ca(2+) moderately activated enzyme activity, where as Zn(2+), Fe(2+) and Hg(2+) inhibited enzyme activity. The protease produced was stable in presence of 50 % (v/v) ethylacetate and acetone. Isopropanol, methanol and benzene increased protease activity by 2.7, 1.3 and 1.1 fold respectively but was inhibited in presence of glycerol and DMSO. This organic solvent-stable protease could be used as a biocatalyst for enzymatic peptide synthesis |
format | Text |
id | pubmed-3048348 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2010 |
publisher | Bentham Open |
record_format | MEDLINE/PubMed |
spelling | pubmed-30483482011-03-04 Purification of Protease from Pseudomonas thermaerum GW1 Isolated from Poultry Waste Site Gaur, Smriti Agrahari, Sarita Wadhwa, Neeraj Open Microbiol J Article An extracellular protease was purified from Pseudomonas thermaerum GW1 a new strain identified by morphological, biochemical and 16S rDNA sequencing. It was isolated from soil of Poultry waste site at Ghazipur near Ghaziabad, Delhi. The strain produces extra cellular protease in the culture media that was maintained at 37°C, 140 rpm. The media was harvested for protease after 48 hrs of incubation at 37°C in basal media supplemented with 1% casein. We report 6.08 fold purification of enzyme following ammonium sulphate precipitation and DEAE-cellulose chromatography. The molecular weight of the enzyme was estimated to be approximately 43,000 daltons as shown by casein zymography studies. The optimum pH for the proteolytic activity was pH 8.0 and enzyme remained stable between pH 5 -11 at 60°C. Interestingly Mn(2+) (5mM) activated enzyme activity by 5 fold, while Cu(2+), Mg(2+)and Ca(2+) moderately activated enzyme activity, where as Zn(2+), Fe(2+) and Hg(2+) inhibited enzyme activity. The protease produced was stable in presence of 50 % (v/v) ethylacetate and acetone. Isopropanol, methanol and benzene increased protease activity by 2.7, 1.3 and 1.1 fold respectively but was inhibited in presence of glycerol and DMSO. This organic solvent-stable protease could be used as a biocatalyst for enzymatic peptide synthesis Bentham Open 2010-08-13 /pmc/articles/PMC3048348/ /pubmed/21379398 http://dx.doi.org/10.2174/1874285801004010067 Text en © Gaur et al.; Licensee Bentham Open. http://creativecommons.org/licenses/by-nc/3.0/ This is an open access article licensed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted, non-commercial use, distribution and reproduction in any medium, provided the work is properly cited. |
spellingShingle | Article Gaur, Smriti Agrahari, Sarita Wadhwa, Neeraj Purification of Protease from Pseudomonas thermaerum GW1 Isolated from Poultry Waste Site |
title | Purification of Protease from Pseudomonas thermaerum GW1 Isolated from Poultry Waste Site |
title_full | Purification of Protease from Pseudomonas thermaerum GW1 Isolated from Poultry Waste Site |
title_fullStr | Purification of Protease from Pseudomonas thermaerum GW1 Isolated from Poultry Waste Site |
title_full_unstemmed | Purification of Protease from Pseudomonas thermaerum GW1 Isolated from Poultry Waste Site |
title_short | Purification of Protease from Pseudomonas thermaerum GW1 Isolated from Poultry Waste Site |
title_sort | purification of protease from pseudomonas thermaerum gw1 isolated from poultry waste site |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3048348/ https://www.ncbi.nlm.nih.gov/pubmed/21379398 http://dx.doi.org/10.2174/1874285801004010067 |
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