Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Cloning, Expression, and Characterization of a Highly Active Alkaline Pectate Lyase from Alkaliphilic Bacillus sp. N16-5

  • Li, Gang (State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences) ;
  • Rao, Lang (State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences) ;
  • Xue, Yanfen (State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences) ;
  • Zhou, Cheng (State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences) ;
  • Zhang, Yun (National Key Lab of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences) ;
  • Ma, Yanhe (State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences)
  • Received : 2009.11.14
  • Accepted : 2009.12.29
  • Published : 2010.04.28
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Abstract

An alkaline pectate lyase, Bsp165PelA, was purified to homogeneity from the culture broth of alkaliphilic Bacillus sp. N16-5. The enzyme showed a specific activity as high as 1,000 U/mg and had optimum activity at pH 11.5 and $50^{\circ}C$. It was composed of a single polypeptide chain with a molecular mass of 42 kDa deduced from SDS-PAGE, and its isoelectric point was around pH 6.0. It could efficiently depolymerize polygalacturonate and pectin. Characterization of product formation revealed unsaturated digalacturonate and trigalacturonate as the main products. The pectate lyase gene (pelA) contained an open reading frame (ORF) of 1,089 bp, encoding a 36-amino acids signal peptide and a mature protein of 326 amino acids with a calculated molecular mass of 35.943 Da. The deduced amino acid sequence from the pelA ORF exhibited significant homology to those of known pectate lyases in polysaccharide lyase family 1. Some conserved active-site amino acids were found in the deduced amino acid sequence of Bsp165PelA. $Ca^{2+}$ was not required for activity on pectic substrates.

Keywords

References

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