Cargando…
Study the effect of F17S mutation on the chimeric Bacillus thermocatenulatus lipase
Lipases (triacylglycerol acylhydrolase, EC 3.1.1.3) are one of the highest value commercial enzymes as they have potential applications in biotechnology for detergents, food, pharmaceuticals, leather, textiles, cosmetics, and paper industries; and are currently receiving considerable attention becau...
Autores principales: | , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Academy of Scientific Research and Technology, Egypt
2016
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6299889/ https://www.ncbi.nlm.nih.gov/pubmed/30647601 http://dx.doi.org/10.1016/j.jgeb.2016.08.002 |
_version_ | 1783381580162531328 |
---|---|
author | Khaleghinejad, Seyed Hossein Motalleb, Gholamreza Karkhane, Ali Asghar Aminzadeh, Saeed Yakhchali, Bagher |
author_facet | Khaleghinejad, Seyed Hossein Motalleb, Gholamreza Karkhane, Ali Asghar Aminzadeh, Saeed Yakhchali, Bagher |
author_sort | Khaleghinejad, Seyed Hossein |
collection | PubMed |
description | Lipases (triacylglycerol acylhydrolase, EC 3.1.1.3) are one of the highest value commercial enzymes as they have potential applications in biotechnology for detergents, food, pharmaceuticals, leather, textiles, cosmetics, and paper industries; and are currently receiving considerable attention because of their potential applications in biotechnology. Bacillus thermocatenulatus Lipase 2 (BTL2) is one of the most important research targets, because of its potential industrial applications. In this study, the effect of substitution Phe17 with Ser in mutated BTL2 lipase, which conserved pentapeptide ((112)Ala-His-Ser-Gln-Gly(116)) was replaced with similar sequences ((207)Gly-Glu-Ser-Ala-Gly(211)) of Candida rugosa lipase (CLR) at the nucleophilic elbow region. Docking results confirmed the mutated lipase to be better than the chimeric lipase. So, cloning was conducted, and the mutated and chimeric btl2 genes were expressed in Escherichia coli, and then the enzymes were purified by anion exchange chromatography. The mutation increased lipase lipolytic activity against most of the applied substrates, with the exception of tributyrin when compared with chimeric lipase. Further, the mutated lipase exhibited higher activity than the chimeric lipase at all temperatures. Optimum pH of the mutated lipase was obtained at pH 9.5, which was more than the chimeric one. Enzyme activity of the mutated lipase in the presence of organic solvents, detergents, and metal ions was also improved than the chimeric lipase. |
format | Online Article Text |
id | pubmed-6299889 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2016 |
publisher | Academy of Scientific Research and Technology, Egypt |
record_format | MEDLINE/PubMed |
spelling | pubmed-62998892019-01-15 Study the effect of F17S mutation on the chimeric Bacillus thermocatenulatus lipase Khaleghinejad, Seyed Hossein Motalleb, Gholamreza Karkhane, Ali Asghar Aminzadeh, Saeed Yakhchali, Bagher J Genet Eng Biotechnol II : Microbial Biotechnology Lipases (triacylglycerol acylhydrolase, EC 3.1.1.3) are one of the highest value commercial enzymes as they have potential applications in biotechnology for detergents, food, pharmaceuticals, leather, textiles, cosmetics, and paper industries; and are currently receiving considerable attention because of their potential applications in biotechnology. Bacillus thermocatenulatus Lipase 2 (BTL2) is one of the most important research targets, because of its potential industrial applications. In this study, the effect of substitution Phe17 with Ser in mutated BTL2 lipase, which conserved pentapeptide ((112)Ala-His-Ser-Gln-Gly(116)) was replaced with similar sequences ((207)Gly-Glu-Ser-Ala-Gly(211)) of Candida rugosa lipase (CLR) at the nucleophilic elbow region. Docking results confirmed the mutated lipase to be better than the chimeric lipase. So, cloning was conducted, and the mutated and chimeric btl2 genes were expressed in Escherichia coli, and then the enzymes were purified by anion exchange chromatography. The mutation increased lipase lipolytic activity against most of the applied substrates, with the exception of tributyrin when compared with chimeric lipase. Further, the mutated lipase exhibited higher activity than the chimeric lipase at all temperatures. Optimum pH of the mutated lipase was obtained at pH 9.5, which was more than the chimeric one. Enzyme activity of the mutated lipase in the presence of organic solvents, detergents, and metal ions was also improved than the chimeric lipase. Academy of Scientific Research and Technology, Egypt 2016-06 2016-09-03 /pmc/articles/PMC6299889/ /pubmed/30647601 http://dx.doi.org/10.1016/j.jgeb.2016.08.002 Text en © 2016 Production and hosting by Elsevier B.V. on behalf of Academy of Scientific Research & Technology. 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 | II : Microbial Biotechnology Khaleghinejad, Seyed Hossein Motalleb, Gholamreza Karkhane, Ali Asghar Aminzadeh, Saeed Yakhchali, Bagher Study the effect of F17S mutation on the chimeric Bacillus thermocatenulatus lipase |
title | Study the effect of F17S mutation on the chimeric Bacillus thermocatenulatus lipase |
title_full | Study the effect of F17S mutation on the chimeric Bacillus thermocatenulatus lipase |
title_fullStr | Study the effect of F17S mutation on the chimeric Bacillus thermocatenulatus lipase |
title_full_unstemmed | Study the effect of F17S mutation on the chimeric Bacillus thermocatenulatus lipase |
title_short | Study the effect of F17S mutation on the chimeric Bacillus thermocatenulatus lipase |
title_sort | study the effect of f17s mutation on the chimeric bacillus thermocatenulatus lipase |
topic | II : Microbial Biotechnology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6299889/ https://www.ncbi.nlm.nih.gov/pubmed/30647601 http://dx.doi.org/10.1016/j.jgeb.2016.08.002 |
work_keys_str_mv | AT khaleghinejadseyedhossein studytheeffectoff17smutationonthechimericbacillusthermocatenulatuslipase AT motallebgholamreza studytheeffectoff17smutationonthechimericbacillusthermocatenulatuslipase AT karkhanealiasghar studytheeffectoff17smutationonthechimericbacillusthermocatenulatuslipase AT aminzadehsaeed studytheeffectoff17smutationonthechimericbacillusthermocatenulatuslipase AT yakhchalibagher studytheeffectoff17smutationonthechimericbacillusthermocatenulatuslipase |