Cloning and overexpression in Escherichia coli of the gene encoding dihydroxyacetone kinase isoenzyme I from Schizosaccharomyces pombe, and its application to dihydroxyacetone phosphate production.
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.
Itoh
N
N
Biotechnology Research Center, Toyama Prefectural University, Japan. itoh@pu-toyama.ac.jp
Tujibata
Y
Y
Liu
J Q
JQ
D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D004098:Dihydroxyacetone / Q000378:metabolism; D004099:Dihydroxyacetone Phosphate / Q000096:biosynthesis*; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics* / Q000378:metabolism; D005990:Glycerol / Q000378:metabolism; D007527:Isoenzymes / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism; D012568:Schizosaccharomyces / Q000201:enzymology* / Q000235:genetics; D017422:Sequence Analysis, DNA
D007527:Isoenzymes; D011994:Recombinant Proteins; D004099:Dihydroxyacetone Phosphate; D017853:Phosphotransferases (Alcohol Group Acceptor); C023132:glycerone kinase; D004098:Dihydroxyacetone; D005990:Glycerol