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Molecular and General Genetics MGG

, Volume 261, Issue 2, pp 236–241 | Cite as

Cloning of the Escherichia coli endo-1,4-D-glucanase gene and identification of its product

  • Y. W. Park
  • H. D. Yun
ORIGINAL PAPER

Abstract

A plasmid (pYP17) containing a genomic DNA insert from Escherichia coli K-12 that confers the ability to hydrolyze carboxymethylcellulose (CMC) was isolated from a genomic library constructed in the cosmid vector pLAFR3 in E. coli DH5α. A small 1.65-kb fragment, designated bcsC (pYP300), was sequenced and found to contain an ORF of 1,104 bp encoding a protein of 368 amino acid residues, with a calculated molecular weight of 41,700 Da. BcsC carries a typical prokaryotic signal peptide of 21 amino acid residues. The predicted amino acid sequence of the BcsC protein is similar to that of CelY of Erwinia chrysanthemi, CMCase of Cellulomonas uda, EngX of Acetobacter xylinum, and CelC of Agrobacterium tumefaciens. Based on these sequence similarities, we propose that the bcsC gene is a member of glycosyl hydrolase family 8. The apparent molecular mass of the protein, when expressed in E. coli, is approximately 40 kDa, and the CMCase activity is found mainly in the extracellular space. The enzyme is optimally active at pH 7 and a temperature of 40° C.

Key wordsEscherichia coli β-1 4-D-glucan glucanohydrolase bcsC Activity staining 

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Copyright information

© Springer-Verlag Berlin Heidelberg 1999

Authors and Affiliations

  • Y. W. Park
    • 1
  • H. D. Yun
    • 2
  1. 1.Genetic Engineering Institute Gyeongsang National University, Chinju 660-701, KoreaKR
  2. 2.Microbial Genetics Laboratory Department of Agricultural Chemistry Gyeongsang National University Chinju 660-701, Korea e-mail: hdyun@nongae.gsnu.ac.kr Tel.: +82-591-7515469; Fax: +82-591-570178KR

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