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Cold-active extracellular lipase: Expression in Sf9 insect cells, purification, and catalysis
Cold-active lipases are gaining special attention nowadays as they are increasingly used in various industries such as fine chemical synthesis, food processing, and washer detergent. In the present study, an extracellular lipase gene from Yarrowia lipolytica (LIPY8) was cloned and expressed by bacul...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Elsevier
2018
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6290134/ https://www.ncbi.nlm.nih.gov/pubmed/30568889 http://dx.doi.org/10.1016/j.btre.2018.e00295 |
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author | Li, Tang Zhang, Wenfa Hao, Jianhua Sun, Mi Lin, Sheng-Xiang |
author_facet | Li, Tang Zhang, Wenfa Hao, Jianhua Sun, Mi Lin, Sheng-Xiang |
author_sort | Li, Tang |
collection | PubMed |
description | Cold-active lipases are gaining special attention nowadays as they are increasingly used in various industries such as fine chemical synthesis, food processing, and washer detergent. In the present study, an extracellular lipase gene from Yarrowia lipolytica (LIPY8) was cloned and expressed by baculovirus expression system. The recombinant lipase (LipY8p) was purified using chromatographic techniques, resulting in a purification factor of 25.7-fold with a specific activity of 1102.9U/mg toward olive oil. The apparent molecular mass of purified LipY8p was 40 kDa. The enzyme was most active at pH 7.5 and 17 °C. It exhibited maximum activity toward medium chain (C10) esters. The presence of transition metals such as Zn(2+), Cu(2+), and Ni(2+) strongly inhibited the enzyme activity, which was enhanced by EDTA. The lipase activity was affected by detergents and was elevated by various organic solvents at 10% (v/v). These enzymatic properties make this lipase of considerable potential for biotechnological applications. |
format | Online Article Text |
id | pubmed-6290134 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | Elsevier |
record_format | MEDLINE/PubMed |
spelling | pubmed-62901342018-12-19 Cold-active extracellular lipase: Expression in Sf9 insect cells, purification, and catalysis Li, Tang Zhang, Wenfa Hao, Jianhua Sun, Mi Lin, Sheng-Xiang Biotechnol Rep (Amst) Article Cold-active lipases are gaining special attention nowadays as they are increasingly used in various industries such as fine chemical synthesis, food processing, and washer detergent. In the present study, an extracellular lipase gene from Yarrowia lipolytica (LIPY8) was cloned and expressed by baculovirus expression system. The recombinant lipase (LipY8p) was purified using chromatographic techniques, resulting in a purification factor of 25.7-fold with a specific activity of 1102.9U/mg toward olive oil. The apparent molecular mass of purified LipY8p was 40 kDa. The enzyme was most active at pH 7.5 and 17 °C. It exhibited maximum activity toward medium chain (C10) esters. The presence of transition metals such as Zn(2+), Cu(2+), and Ni(2+) strongly inhibited the enzyme activity, which was enhanced by EDTA. The lipase activity was affected by detergents and was elevated by various organic solvents at 10% (v/v). These enzymatic properties make this lipase of considerable potential for biotechnological applications. Elsevier 2018-12-04 /pmc/articles/PMC6290134/ /pubmed/30568889 http://dx.doi.org/10.1016/j.btre.2018.e00295 Text en © 2018 The Authors http://creativecommons.org/licenses/by-nc-nd/4.0/ This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/). |
spellingShingle | Article Li, Tang Zhang, Wenfa Hao, Jianhua Sun, Mi Lin, Sheng-Xiang Cold-active extracellular lipase: Expression in Sf9 insect cells, purification, and catalysis |
title | Cold-active extracellular lipase: Expression in Sf9 insect cells, purification, and catalysis |
title_full | Cold-active extracellular lipase: Expression in Sf9 insect cells, purification, and catalysis |
title_fullStr | Cold-active extracellular lipase: Expression in Sf9 insect cells, purification, and catalysis |
title_full_unstemmed | Cold-active extracellular lipase: Expression in Sf9 insect cells, purification, and catalysis |
title_short | Cold-active extracellular lipase: Expression in Sf9 insect cells, purification, and catalysis |
title_sort | cold-active extracellular lipase: expression in sf9 insect cells, purification, and catalysis |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6290134/ https://www.ncbi.nlm.nih.gov/pubmed/30568889 http://dx.doi.org/10.1016/j.btre.2018.e00295 |
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