Cloning and characterization of a new cold-active lipase from a deep-sea sediment metagenome SCIE SCOPUS

Cited 112 time in WEB OF SCIENCE Cited 128 time in Scopus
Title
Cloning and characterization of a new cold-active lipase from a deep-sea sediment metagenome
Author(s)
Jeon, Jeong Ho; Kim, Jun Tae; Kim, Yun Jae; Kim, Hyung-Kwoun; Lee, Hyun Sook; Kang, Sung Gyun; Kim, Sang-Jin; Lee, Jung-Hyun
KIOST Author(s)
Lee, Hyun Sook(이현숙)Kang, Sung Gyun(강성균)Lee, Jung Hyun(이정현)
Publication Year
2009-01
Abstract
To search for new cold-active lipases, a metagenomic library was constructed using cold-sea sediment samples at Edison Seamount and was screened for lipolytic activities by plating on a tricaprylin medium. Subsequently, a fosmid clone was selected, and the whole sequence of 36 kb insert of the fosmid clone was determined by shotgun sequencing. The sequence analysis revealed the presence of 25 open reading frames (ORF), and ORF20 (EML1) showed similarities to lipases. Phylogenetic analysis of EML1 suggested that the protein belonged to a new family of esterase/lipase together with LipG. The EML1 gene was expressed in Escherichia coli, and purified by metal-chelating chromatography. The optimum activity of the purified EML1 (rEML1) occurred at pH 8.0 and 25A degrees C, respectively, and rEML1 displayed more than 50% activity at 5A degrees C. The activation energy for the hydrolysis of olive oil was determined to be 3.28 kcal/mol, indicating that EML1 is a cold-active lipase. rEML1 preferentially hydrolyzed triacylglycerols acyl-group chains with long chain lengths of a parts per thousand yen8 carbon atoms and displayed hydrolyzing activities toward various natural oil substrates. rEML1 was resistant to various detergents such as Triton X-100 and Tween 80. This study represents an example which developed a new cold-active lipase from a deep-sea sediment metagenome.
ISSN
0175-7598
URI
https://sciwatch.kiost.ac.kr/handle/2020.kiost/4343
DOI
10.1007/s00253-008-1656-2
Bibliographic Citation
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY, v.81, no.5, pp.865 - 874, 2009
Publisher
SPRINGER
Keywords
Lipase; Metagenome; Deep-sea sediment
Type
Article
Language
English
Document Type
Article
Publisher
SPRINGER
Related Researcher
Research Interests

marine biotechnology,molecular microbiology,해양생명공학,분자미생물학

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