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Purification, characterisation and expression in Saccharomyces cerevisiae of LipG7 an enantioselective, cold-adapted lipase from the Antarctic filamentous fungus Geomyces sp. P7 with unusual thermostability characteristics

Paper ID Volume ID Publish Year Pages File Format Full-Text
17175 42649 2013 7 PDF Available
Title
Purification, characterisation and expression in Saccharomyces cerevisiae of LipG7 an enantioselective, cold-adapted lipase from the Antarctic filamentous fungus Geomyces sp. P7 with unusual thermostability characteristics
Abstract

•LipG7, an enantioselective, cold-adapted lipase was purified.•LipG7 retains 100% of initial activity after thermal denaturation.•LipG7 effectively catalysed the enantioselective transesterification of a secondary alcohol.•Low coverage, high throughput transcriptome sequencing identified the LipG7 gene.•Heterologously expressed LipG7 retains its thermostability characteristics.

A lipase, LipG7, has been purified from the Antarctic filamentous fungus Geomyces sp. P7 which was found to be cold-adapted and able to retain/regain its activity after heat denaturation. The LipG7 exhibits 100% residual activity following 1 h incubation at 100 °C whilst simultaneously showing kinetic adaptations to cold temperatures. LipG7 was also found to have industrial potential as an enantioselective biocatalyst as it is able to effectively catalyse the enantioselective transesterification of a secondary alcohol. The LipG7 coding sequence has been identified and cloned using 454 pyrosequencing of the transcriptome and inverse PCR. The LipG7 protein has been heterologously expressed in Saccharomyces cerevisiae BJ5465 and shown to exhibit the same characteristics as the native protein.

Keywords
PMSF, phenylmethylsulphonyl fluoride; CalB, Candida antarctica lipase BCold adaptation; Thermostability; Lipase; Psychrophillic enzyme; Inverse-PCR
First Page Preview
Purification, characterisation and expression in Saccharomyces cerevisiae of LipG7 an enantioselective, cold-adapted lipase from the Antarctic filamentous fungus Geomyces sp. P7 with unusual thermostability characteristics
Publisher
Database: Elsevier - ScienceDirect
Journal: Enzyme and Microbial Technology - Volume 53, Issue 1, 10 June 2013, Pages 18–24
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering