Abstract
The gene encoding sucrose phosphorylase (742sp) in Leuconostoc mesenteroides NRRL B-742 was cloned and expressed in Escherichia coli. The nucleotide sequence of the transformed 742sp comprised an ORF of 1,458 bp giving a protein with calculated molecular mass of 55.3 kDa. 742SPase contains a C-terminal amino acid sequence that is significantly different from those of other Leu. mesenteroides SPases. The purified 742SPase had a specific activity of 1.8 U/mg with a K (m) of 3 mM with sucrose as a substrate; optimum activity was at 37 degrees C and pH 6.7. The purified 742SPase transferred the glucosyl moiety of sucrose to cytosine monophosphate (CMP).
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Amino Acid Sequence
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Bacterial Proteins / chemistry
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Bacterial Proteins / genetics
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Bacterial Proteins / metabolism*
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Chromatography, Thin Layer
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Cloning, Molecular / methods
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Cytidine Monophosphate / metabolism
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Electrophoresis, Polyacrylamide Gel
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Escherichia coli / genetics*
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Gene Expression
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Glucosyltransferases / chemistry
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Glucosyltransferases / genetics
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Glucosyltransferases / metabolism*
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Leuconostoc / enzymology*
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Leuconostoc / genetics
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Molecular Sequence Data
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Molecular Structure
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Recombinant Proteins / isolation & purification
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Recombinant Proteins / metabolism
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Sequence Homology, Amino Acid
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Substrate Specificity
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Sucrose / metabolism
Substances
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Bacterial Proteins
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Recombinant Proteins
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Sucrose
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Glucosyltransferases
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sucrose phosphorylase
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Cytidine Monophosphate