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