Abstract
A DNA fragment, which complemented the growth of E. coli both on M9 medium containing raffinose and on LB medium containing ampicillin, IPTG and 5-bromo-4-chloro-3-indoxyl-alpha-D-galactoside, was isolated from the genomic library of Bifidobacterium longum SJ32, which had been digested with EcoRI. In the cloned DNA fragment, a gene encoding a sucrose phosphorylase (splP) and a partially cloned putative sucrose regulator gene (splR) were identified using the deletion analysis and sequence analysis. A 56 kDa protein was synthesized in E. coli and partially purified by DEAE-ion exchange chromatography. The partially purified enzyme did not react with melibiose, melezitoze and raffinose but did with sucrose. It had transglucosylation activity in addition to hydrolytic activity.
Publication types
-
Research Support, Non-U.S. Gov't
MeSH terms
-
Amino Acid Sequence
-
Bifidobacterium / chemistry
-
Bifidobacterium / enzymology*
-
Bifidobacterium / genetics*
-
Cloning, Molecular
-
Enzyme Activation
-
Escherichia coli / chemistry
-
Escherichia coli / enzymology*
-
Escherichia coli / genetics*
-
Gene Expression Regulation, Bacterial / physiology
-
Gene Expression Regulation, Enzymologic / physiology
-
Glucosyltransferases / biosynthesis*
-
Glucosyltransferases / chemistry*
-
Glucosyltransferases / genetics
-
Molecular Sequence Data
-
Oligosaccharides / chemistry*
-
Oligosaccharides / classification
-
Protein Engineering / methods*
-
Recombinant Proteins / chemistry
-
Recombinant Proteins / metabolism
-
Substrate Specificity
Substances
-
Oligosaccharides
-
Recombinant Proteins
-
Glucosyltransferases
-
sucrose phosphorylase