Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Selection and Characterization of Forest Soil Metagenome Genes Encoding Lipolytic Enzymes

  • Hong, Kyung-Sik (Bio-Organic Science Division, Korea Research Institute of Chemical Technology) ;
  • Lim, He-Kyoung (Bio-Organic Science Division, Korea Research Institute of Chemical Technology) ;
  • Chung, Eu-Jin (Division of Applied Biology, College of Natural Resources and Life Science, Dong-A University) ;
  • Park, Eun-Jin (Division of Applied Biology, College of Natural Resources and Life Science, Dong-A University) ;
  • Lee, Myung-Hwan (Division of Applied Biology, College of Natural Resources and Life Science, Dong-A University) ;
  • Kim, Jin-Cheol (Bio-Organic Science Division, Korea Research Institute of Chemical Technology) ;
  • Cho, Gyung-Ja (Bio-Organic Science Division, Korea Research Institute of Chemical Technology) ;
  • Cho, Kwang-Yun (Bio-Organic Science Division, Korea Research Institute of Chemical Technology) ;
  • Lee, Seon-Woo (Division of Applied Biology, College of Natural Resources and Life Science, Dong-A University)
  • Published : 2007.10.30
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Abstract

A metagenome is a unique resource to search for novel microbial enzymes from the unculturable microorganisms in soil. A forest soil metagenomic library using a fosmid and soil microbial DNA from Gwangneung forest, Korea, was constructed in Escherichia coli and screened to select lipolytic genes. A total of seven unique lipolytic clones were selected by screening of the 31,000-member forest soil metagenome library based on tributyrin hydrolysis. The ORFs for lipolytic activity were subcloned in a high copy number plasmid by screening the secondary shortgun libraries from the seven clones. Since the lipolytic enzymes were well secreted in E. coli into the culture broth, the lipolytic activity of the subclones was confirmed by the hydrolysis of p-nitrophenyl butyrate using culture supernatant. Deduced amino acid sequence analysis of the identified ORFs for lipolytic activity revealed that 4 genes encode hormone-sensitive lipase (HSL) in lipase family IV. Phylogenetic analysis indicated that 4 proteins were clustered with HSL in the database and other metagenomic HSLs. The other 2 genes and 1 gene encode non-heme peroxidase-like enzymes of lipase family V and a GDSL family esterase/lipase in family II, respectively. The gene for the GDSL enzyme is the first description of the enzyme from metagenomic screening.

Keywords

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