Abstract
The gene dak1 encoding a dihydroxyacetone kinase (DHAK) isoenzyme I, one of two isoenzymes in the Schizosaccharomyces pombe IFO 0354 strain, was cloned and sequenced. The dak1 gene comprises 1743 bp and encodes a protein of 62,245 Da. The deduced amino acid sequence showed a similarity to a putative DHAK of Saccharomyces cerevisiae and DHAK of Citrobacter freundii. The dak1 gene was expressed at a high level in Escherichia coli, and the recombinant enzyme was purified to homogeneity and characterized. The acetone powder of recombinant E. coli cells was used to produce dihydroxyacetone phosphate.
MeSH terms
-
Amino Acid Sequence
-
Base Sequence
-
Cloning, Molecular
-
Dihydroxyacetone / metabolism
-
Dihydroxyacetone Phosphate / biosynthesis*
-
Electrophoresis, Polyacrylamide Gel
-
Escherichia coli / enzymology
-
Escherichia coli / genetics*
-
Escherichia coli / metabolism
-
Glycerol / metabolism
-
Isoenzymes / genetics
-
Isoenzymes / metabolism
-
Molecular Sequence Data
-
Phosphotransferases (Alcohol Group Acceptor) / chemistry
-
Phosphotransferases (Alcohol Group Acceptor) / genetics*
-
Phosphotransferases (Alcohol Group Acceptor) / metabolism*
-
Recombinant Proteins / metabolism
-
Schizosaccharomyces / enzymology*
-
Schizosaccharomyces / genetics
-
Sequence Analysis, DNA
Substances
-
Isoenzymes
-
Recombinant Proteins
-
Dihydroxyacetone Phosphate
-
Phosphotransferases (Alcohol Group Acceptor)
-
glycerone kinase
-
Dihydroxyacetone
-
Glycerol