ORIGINAL PAPER

Applied Microbiology and Biotechnology

, Volume 51, Issue 2, pp 193-200

Cloning and overexpression in Escherichia coli of the gene encoding dihydroxyacetone kinase isoenzyme I from Schizosaccharomyces pombe, and its application to dihydroxyacetone phosphate production

  • N. ItohAffiliated withBiotechnology Research Center, Toyama Prefectural University, 5180 Kurokawa Kosugi, Toyama 939-0398, Japan e-mail: itoh@pu-toyama.ac.jp Tel.: +81-766-56-7500, ext. 560 Fax: +81-766-56-2498
  • , Y. TujibataAffiliated withDepartment of Applied Chemistry and Biotechnology, Faculty of Engineering, Fukui University, Bunkyo 3-9-1, Fukui 910-8507, Japan
  • , J. Q. LiuAffiliated withBiotechnology Research Center, Toyama Prefectural University, 5180 Kurokawa Kosugi, Toyama 939-0398, Japan e-mail: itoh@pu-toyama.ac.jp Tel.: +81-766-56-7500, ext. 560 Fax: +81-766-56-2498

Rent the article at a discount

Rent now

* Final gross prices may vary according to local VAT.

Get Access

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.