Gene cloning, heterologous expression, and partial characterization of a novel cold-adapted subfamily I.3 lipase from Pseudomonas fluorescence KE38

Abstract A novel cold-active true lipase from Pseudomonas sp. KE38 was cloned, sequencing and expressed in E. coli by degenerate PCR and genome walking technique. The open reading frame of the cloned gene encoded a polypeptide chain of 617 amino acids with a confirmed molecular weight of 64 kD. Phyl...

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Autores principales:	Fulya Karakaş, Alper Arslanoğlu
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2020
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/81cd23fa1cb3472c866b54caf496441a
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spelling	oai:doaj.org-article:81cd23fa1cb3472c866b54caf496441a2021-12-02T13:34:00ZGene cloning, heterologous expression, and partial characterization of a novel cold-adapted subfamily I.3 lipase from Pseudomonas fluorescence KE3810.1038/s41598-020-79199-w2045-2322https://doaj.org/article/81cd23fa1cb3472c866b54caf496441a2020-12-01T00:00:00Zhttps://doi.org/10.1038/s41598-020-79199-whttps://doaj.org/toc/2045-2322Abstract A novel cold-active true lipase from Pseudomonas sp. KE38 was cloned, sequencing and expressed in E. coli by degenerate PCR and genome walking technique. The open reading frame of the cloned gene encoded a polypeptide chain of 617 amino acids with a confirmed molecular weight of 64 kD. Phylogenetic analysis of the deduced amino acid sequence of the lipase indicated that it had high similarity with lipases of subfamily Ι.3 of bacterial lipases. Recombinant lipase was purified in denatured form as inclusion bodies, which were then renatured by urea followed by dialysis. Lipase activity was determined titrimetrically using olive oil as substrate. The enzyme showed optimal activity at 25 °C, pH 8.5 and was highly stable in the presence of various metal ions and organic solvents. Low optimal temperature and high activity in the presence of methanol and ethanol make this lipase a potential candidate for transesterification reactions and biodiesel production.Fulya KarakaşAlper ArslanoğluNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 10, Iss 1, Pp 1-13 (2020)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Fulya Karakaş Alper Arslanoğlu Gene cloning, heterologous expression, and partial characterization of a novel cold-adapted subfamily I.3 lipase from Pseudomonas fluorescence KE38
description	Abstract A novel cold-active true lipase from Pseudomonas sp. KE38 was cloned, sequencing and expressed in E. coli by degenerate PCR and genome walking technique. The open reading frame of the cloned gene encoded a polypeptide chain of 617 amino acids with a confirmed molecular weight of 64 kD. Phylogenetic analysis of the deduced amino acid sequence of the lipase indicated that it had high similarity with lipases of subfamily Ι.3 of bacterial lipases. Recombinant lipase was purified in denatured form as inclusion bodies, which were then renatured by urea followed by dialysis. Lipase activity was determined titrimetrically using olive oil as substrate. The enzyme showed optimal activity at 25 °C, pH 8.5 and was highly stable in the presence of various metal ions and organic solvents. Low optimal temperature and high activity in the presence of methanol and ethanol make this lipase a potential candidate for transesterification reactions and biodiesel production.
format	article
author	Fulya Karakaş Alper Arslanoğlu
author_facet	Fulya Karakaş Alper Arslanoğlu
author_sort	Fulya Karakaş
title	Gene cloning, heterologous expression, and partial characterization of a novel cold-adapted subfamily I.3 lipase from Pseudomonas fluorescence KE38
title_short	Gene cloning, heterologous expression, and partial characterization of a novel cold-adapted subfamily I.3 lipase from Pseudomonas fluorescence KE38
title_full	Gene cloning, heterologous expression, and partial characterization of a novel cold-adapted subfamily I.3 lipase from Pseudomonas fluorescence KE38
title_fullStr	Gene cloning, heterologous expression, and partial characterization of a novel cold-adapted subfamily I.3 lipase from Pseudomonas fluorescence KE38
title_full_unstemmed	Gene cloning, heterologous expression, and partial characterization of a novel cold-adapted subfamily I.3 lipase from Pseudomonas fluorescence KE38
title_sort	gene cloning, heterologous expression, and partial characterization of a novel cold-adapted subfamily i.3 lipase from pseudomonas fluorescence ke38
publisher	Nature Portfolio
publishDate	2020
url	https://doaj.org/article/81cd23fa1cb3472c866b54caf496441a
work_keys_str_mv	AT fulyakarakas genecloningheterologousexpressionandpartialcharacterizationofanovelcoldadaptedsubfamilyi3lipasefrompseudomonasfluorescenceke38 AT alperarslanoglu genecloningheterologousexpressionandpartialcharacterizationofanovelcoldadaptedsubfamilyi3lipasefrompseudomonasfluorescenceke38
_version_	1718392805310595072

Gene cloning, heterologous expression, and partial characterization of a novel cold-adapted subfamily I.3 lipase from Pseudomonas fluorescence KE38

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