Nucleotide sequence of the Clostridium stercorarium xynB gene encoding an extremely thermostable xylanase, and characterization of the translated product

Biosci Biotechnol Biochem. 1995 Jan;59(1):40-6. doi: 10.1271/bbb.59.40.

Abstract

The nucleotides of the xynB gene of Clostridium stercorarium F-9 were sequenced. The structural gene consists of an open reading frame of 1161 bp encoding a xylanase (XynB) in family F of 387 amino acids with a molecular weight of 44,377. The molecular weight of the enzyme purified from a recombinant Escherichia coli was around 41,000, smaller than the predicted value, on SDS-polyacrylamide gel electrophoresis due to the lack of 32 amino acids at the N-terminus. Intact XynB with a molecular weight of around 43,000 was immunologically detected in the total cell proteins of a recombinant E. coli and C. stercorarium F-9. The purified XynB was active toward xylan, carboxymethylcellulose, p-nitrophenyl-beta-D-xylopyranoside and p-nitrophenyl-beta-D-cellobioside. The pH optimum was 7.0 and it was quite stable over the pH range of 5 to 12 at 4 degrees C. This enzyme was optimally active at 80 degrees C and retained about 50% of the original activity even after incubation at 100 degrees C for 10 min.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Base Sequence
  • Clostridium / enzymology
  • Clostridium / genetics*
  • Electrophoresis, Polyacrylamide Gel
  • Enzyme Stability
  • Escherichia coli / enzymology*
  • Escherichia coli / genetics
  • Gene Expression Regulation, Enzymologic / genetics
  • Glucosides / chemistry
  • Glycoside Hydrolases / chemistry
  • Glycosides / chemistry
  • Hot Temperature
  • Hydrogen-Ion Concentration
  • Molecular Sequence Data
  • Molecular Weight
  • Nitrophenylgalactosides / chemistry
  • Promoter Regions, Genetic
  • Protein Biosynthesis / genetics
  • Restriction Mapping
  • Sequence Homology, Amino Acid
  • Sequence Homology, Nucleic Acid
  • Substrate Specificity
  • Xylan Endo-1,3-beta-Xylosidase
  • Xylosidases / chemistry
  • Xylosidases / genetics*
  • Xylosidases / isolation & purification
  • Xylosidases / metabolism
  • beta-Glucosidase / chemistry

Substances

  • Glucosides
  • Glycosides
  • 4-nitrophenyl beta-D-xyloside
  • Nitrophenylgalactosides
  • 4-nitrophenylgalactoside
  • 4-nitrophenyl beta-cellobioside
  • Glycoside Hydrolases
  • Xylosidases
  • cellobiosidase
  • beta-Glucosidase
  • Xylan Endo-1,3-beta-Xylosidase
  • exo-1,4-beta-D-xylosidase

Associated data

  • GENBANK/D12504