Applied Microbiology and Biotechnology

, Volume 81, Issue 5, pp 865–874

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

  • Jeong Ho Jeon
  • Jun-Tae Kim
  • Yun Jae Kim
  • Hyung-Kwoun Kim
  • Hyun Sook Lee
  • Sung Gyun Kang
  • Sang-Jin Kim
  • Jung-Hyun Lee
Biotechnologically Relevant Enzymes and Proteins

DOI: 10.1007/s00253-008-1656-2

Cite this article as:
Jeon, J.H., Kim, JT., Kim, Y.J. et al. Appl Microbiol Biotechnol (2009) 81: 865. doi:10.1007/s00253-008-1656-2

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 25°C, respectively, and rEML1 displayed more than 50% activity at 5°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 ≥8 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.

Keywords

Lipase Metagenome Deep-sea sediment 

Copyright information

© Springer-Verlag 2008

Authors and Affiliations

  • Jeong Ho Jeon
    • 1
  • Jun-Tae Kim
    • 1
  • Yun Jae Kim
    • 1
  • Hyung-Kwoun Kim
    • 2
  • Hyun Sook Lee
    • 1
  • Sung Gyun Kang
    • 1
  • Sang-Jin Kim
    • 1
  • Jung-Hyun Lee
    • 1
  1. 1.Marine Biotechnology CenterKorea Ocean Research and Development InstituteSeoulSouth Korea
  2. 2.Division of BiotechnologyThe Catholic University of KoreaBucheonSouth Korea

Personalised recommendations