Thermostable alpha-galactosidase from Thermotoga neapolitana: cloning, sequencing and expression

FEMS Microbiol Lett. 1998 Jun 1;163(1):37-42. doi: 10.1111/j.1574-6968.1998.tb13023.x.

Abstract

A gene encoding a thermostable alpha-galactosidase from the hyperthermophile Thermotoga neapolitana was cloned and sequenced. Sequence analysis showed that the 552-amino acid protein is similar to Escherichia coli Raf type alpha-galactosidase and belongs to Family 36 of the glycosyl hydrolases. Recombinant alpha-galactosidase expressed in E. coli has a molecular mass of ca. 61 kDa, and an optimum activity at 93 degrees C at pH 7.0. The enzyme is highly thermostable and retains 75% of activity after heating to 80 degrees C for 4 h. The potential application of the enzyme to high temperature processing of soy molasses has been demonstrated.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Amino Acid Sequence
  • Base Sequence
  • Cloning, Molecular
  • Enzyme Stability
  • Escherichia coli / genetics
  • Genes, Bacterial / genetics*
  • Gram-Negative Anaerobic Bacteria / enzymology
  • Gram-Negative Anaerobic Bacteria / genetics*
  • Hot Temperature
  • Hydrolysis
  • Melibiose / metabolism
  • Molasses
  • Molecular Sequence Data
  • Molecular Weight
  • Recombinant Fusion Proteins
  • Sequence Analysis
  • Sequence Analysis, DNA
  • Sequence Homology, Amino Acid
  • alpha-Galactosidase / chemistry
  • alpha-Galactosidase / genetics*
  • alpha-Galactosidase / isolation & purification

Substances

  • Recombinant Fusion Proteins
  • Melibiose
  • alpha-Galactosidase

Associated data

  • GENBANK/AF011400