A novel endo-glucanase from the thermophilic bacterium Geobacillus sp. 70PC53 with high activity and stability over a broad range of temperatures

Extremophiles. 2009 May;13(3):425-35. doi: 10.1007/s00792-009-0228-4. Epub 2009 Mar 19.

Abstract

A thermophilic Geobacillus bacterium secreting high activity of endo-glucanase (EC 3.2.1.4) was isolated from rice straw compost supplemented with pig manure. A full-length gene of 1,104 bp, celA, encoding this glycosyl hydrolase family 5 endo-glucanase of 368 amino acids was isolated. No related gene from Geobacillus has been reported previously. The recombinant CelA expressed in Escherichia coli had an optimal activity at 65 degrees C and pH 5.0, and it exhibited tenfold greater specific activity than the commercially available Trichoderma reesei endo-glucanase. CelA displayed activity over a broad temperature range from 45 to 75 degrees C and was a thermostable enzyme with 90% activity retained after heating at 65 degrees C for 6 h. Interestingly, CelA activity could be enhanced by 100% in the presence of 2 mM MnSO(4). CelA had high specific activity over beta-D-glucan from barley and Lichenan, making it a potentially useful enzyme in biofuel and food industries.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Bacillus / enzymology*
  • Bacillus / genetics
  • Base Sequence
  • DNA Primers
  • DNA, Ribosomal / genetics
  • Enzyme Stability
  • Escherichia coli / genetics
  • Glycoside Hydrolases / genetics
  • Glycoside Hydrolases / isolation & purification*
  • Glycoside Hydrolases / metabolism
  • Hydrogen-Ion Concentration
  • Molecular Sequence Data
  • Phylogeny
  • RNA, Ribosomal, 16S / genetics
  • Recombinant Proteins / genetics
  • Recombinant Proteins / isolation & purification
  • Recombinant Proteins / metabolism
  • Substrate Specificity
  • Temperature

Substances

  • DNA Primers
  • DNA, Ribosomal
  • RNA, Ribosomal, 16S
  • Recombinant Proteins
  • Glycoside Hydrolases