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Characterization of a Hyperthermostable Alkaline Lipase from Bacillus sonorensis 4R
Hyperthermostable alkaline lipase from Bacillus sonorensis 4R was purified and characterized. The enzyme production was carried out at 80°C and 9.0 pH in glucose-tween inorganic salt broth under static conditions for 96 h. Lipase was purified by anion exchange chromatography by 12.15 fold with a yie...
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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Hindawi Publishing Corporation
2016
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4745284/ https://www.ncbi.nlm.nih.gov/pubmed/26904276 http://dx.doi.org/10.1155/2016/4170684 |
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author | Bhosale, Hemlata Shaheen, Uzma Kadam, Tukaram |
author_facet | Bhosale, Hemlata Shaheen, Uzma Kadam, Tukaram |
author_sort | Bhosale, Hemlata |
collection | PubMed |
description | Hyperthermostable alkaline lipase from Bacillus sonorensis 4R was purified and characterized. The enzyme production was carried out at 80°C and 9.0 pH in glucose-tween inorganic salt broth under static conditions for 96 h. Lipase was purified by anion exchange chromatography by 12.15 fold with a yield of 1.98%. The molecular weight of lipase was found to be 21.87 KDa by SDS-PAGE. The enzyme activity was optimal at 80°C with t (1/2) of 150 min and at 90°C, 100°C, 110°C, and 120°C; the respective values were 121.59 min, 90.01 min, 70.01 min, and 50 min. The enzyme was highly activated by Mg and t (1/2) values at 80°C were increased from 150 min to 180 min when magnesium and mannitol were added in combination. The activation energy calculated from Arrhenius plot was 31.102 KJ/mol. At 80–120°C, values of ΔH and ΔG were in the range of 28.16–27.83 KJ/mol and 102.79 KJ/mol to 111.66 KJ/mol, respectively. Lipase activity was highest at 9.0 pH and stable for 2 hours at this pH at 80°C. Pretreatment of lipase with MgSO(4) and CaSO(4) stimulated enzyme activity by 249.94% and 30.2%, respectively. The enzyme activity was greatly reduced by CoCl(2), CdCl(2), HgCl(2), CuCl(2), Pb(NO(3))(2), PMSF, orlistat, oleic acid, iodine, EDTA, and urea. |
format | Online Article Text |
id | pubmed-4745284 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2016 |
publisher | Hindawi Publishing Corporation |
record_format | MEDLINE/PubMed |
spelling | pubmed-47452842016-02-22 Characterization of a Hyperthermostable Alkaline Lipase from Bacillus sonorensis 4R Bhosale, Hemlata Shaheen, Uzma Kadam, Tukaram Enzyme Res Research Article Hyperthermostable alkaline lipase from Bacillus sonorensis 4R was purified and characterized. The enzyme production was carried out at 80°C and 9.0 pH in glucose-tween inorganic salt broth under static conditions for 96 h. Lipase was purified by anion exchange chromatography by 12.15 fold with a yield of 1.98%. The molecular weight of lipase was found to be 21.87 KDa by SDS-PAGE. The enzyme activity was optimal at 80°C with t (1/2) of 150 min and at 90°C, 100°C, 110°C, and 120°C; the respective values were 121.59 min, 90.01 min, 70.01 min, and 50 min. The enzyme was highly activated by Mg and t (1/2) values at 80°C were increased from 150 min to 180 min when magnesium and mannitol were added in combination. The activation energy calculated from Arrhenius plot was 31.102 KJ/mol. At 80–120°C, values of ΔH and ΔG were in the range of 28.16–27.83 KJ/mol and 102.79 KJ/mol to 111.66 KJ/mol, respectively. Lipase activity was highest at 9.0 pH and stable for 2 hours at this pH at 80°C. Pretreatment of lipase with MgSO(4) and CaSO(4) stimulated enzyme activity by 249.94% and 30.2%, respectively. The enzyme activity was greatly reduced by CoCl(2), CdCl(2), HgCl(2), CuCl(2), Pb(NO(3))(2), PMSF, orlistat, oleic acid, iodine, EDTA, and urea. Hindawi Publishing Corporation 2016 2016-01-21 /pmc/articles/PMC4745284/ /pubmed/26904276 http://dx.doi.org/10.1155/2016/4170684 Text en Copyright © 2016 Hemlata Bhosale et al. https://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Research Article Bhosale, Hemlata Shaheen, Uzma Kadam, Tukaram Characterization of a Hyperthermostable Alkaline Lipase from Bacillus sonorensis 4R |
title | Characterization of a Hyperthermostable Alkaline Lipase from Bacillus sonorensis 4R |
title_full | Characterization of a Hyperthermostable Alkaline Lipase from Bacillus sonorensis 4R |
title_fullStr | Characterization of a Hyperthermostable Alkaline Lipase from Bacillus sonorensis 4R |
title_full_unstemmed | Characterization of a Hyperthermostable Alkaline Lipase from Bacillus sonorensis 4R |
title_short | Characterization of a Hyperthermostable Alkaline Lipase from Bacillus sonorensis 4R |
title_sort | characterization of a hyperthermostable alkaline lipase from bacillus sonorensis 4r |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4745284/ https://www.ncbi.nlm.nih.gov/pubmed/26904276 http://dx.doi.org/10.1155/2016/4170684 |
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