Journal of agriculture & life science (농업생명과학연구)

Institute of Agriculture & Life Science, Gyeongsang National University (경상대학교 농업생명과학연구원)
권호 표지

Cloning and Characterization of Cellulase Gene (cel5B) from Cow Rumen Metagenome

  • Kang, Tae-Ho (Division of Applied Life Science (BK21 Program), Gyeongsang National Univ.) ;
  • Kim, Min-Keun (Gyeongsangnam-do Agricultural Research and Extension Service) ;
  • Barman, Dhirendra Nath (Division of Applied Life Science (BK21 Program), Gyeongsang National Univ.) ;
  • Kim, Jung-Ho (Department of Agricultural Chemistry, Sunchon National University) ;
  • Kim, Hoon (Department of Agricultural Chemistry, Sunchon National University) ;
  • Yun, Han-Dae (Division of Applied Life Science (BK21 Program), Gyeongsang National Univ.)
  • Received : 2012.01.17
  • Accepted : 2012.04.27
  • Published : 2012.04.30
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Abstract

A carboxymethyl cellulase gene, cel5B, was cloned, sequenced, and expressed in Escherichia coli. pRCS20 in E. coli was identified from metagenomic cosmid library of cow rumen for cellulase activity on a carboxymethyl cellulose agar plates. Cosmid clone (RCS20) was partially digested with Sau3AI, ligated into BamHI site of pBluescript II SK+ vector, and transformed into E. coli $DH5{\alpha}$. The insert DNA of 1.3 kb was obtained, designated cel5B, which has the activity of hydrolyzation of CMC. The cel5B gene had an open reading frame (ORF) of 1,059 bp encoding 352 amino acids with a signal peptide of 48 amino acids and the conserved region, VIYEIYNEPL, belongs to the glycosyl hydrolase family 5. The molecular mass of Cel5B protein expressed from E. coli $DH5{\alpha}$ exhibited to be about 34 kDa by CMC-SDS-PAGE. The optimal pH was 8.0, and the optimal temperature was about $50^{\circ}C$ for its enzymatic activity.

Keywords

Acknowledgement

Grant : Cooperative Research Program for Agriculture Science & Technology Development

Supported by : Rural Development Administration

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