Molecular Cloning of a Cellulase Gene from Abalone Haliotis discus hannai and Its Expression in E coli

  • Park, Eun-Mi (Biotechnology Research Institute, National Fisheries Research & Development Institute) ;
  • Han, Yun-Hee (Biotechnology Research Institute, National Fisheries Research & Development Institute) ;
  • Park, In-Suk (Biotechnology Research Institute, National Fisheries Research & Development Institute) ;
  • Nam, Bo-Hye (Biotechnology Research Institute, National Fisheries Research & Development Institute) ;
  • Kong, Hee Jeong (Biotechnology Research Institute, National Fisheries Research & Development Institute) ;
  • Kim, Woo-Jin (Biotechnology Research Institute, National Fisheries Research & Development Institute) ;
  • Lee, Sang-Jun (Biotechnology Research Institute, National Fisheries Research & Development Institute) ;
  • Kim, Young-Ok (Biotechnology Research Institute, National Fisheries Research & Development Institute)
  • Published : 2007.06.30

Abstract

A cellulase (endo-${\beta}$-1,4-D-glucanase(E.C.3.2.1.4)) was isolated from the hepatopancreas of abalone Haliotis discus hannai by EST analysis. The abalone cellulase named HdEG compassed 1977 bp, including 195 bp in the 5'untranslated region, 1680 bp in the open reading frame which encodes 560 amino acid residues, and 92 bp in the 3'-untranslated region. The C-terminal region of the HdEG showed 44-52% identity to the catalytic domains of glycoside hydrolase family 9 (GHF9)-cellulases from arthropods and bacteria. The recombinant cellulase, pEHdEG was produced in E. coli with being fused with C-terminal His-tag. The expressed protein showed a single band (~62 kDa) on Western blotting which was consistent with the value (61,878 Da) calculated from the DNA sequence.

Keywords