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. Itoh
  • Y. Tujibata
  • J. Q. Liu
ORIGINAL PAPER

DOI: 10.1007/s002530051381

Cite this article as:
Itoh, N., Tujibata, Y. & Liu, J. Appl Microbiol Biotechnol (1999) 51: 193. doi:10.1007/s002530051381

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.

Copyright information

© Springer-Verlag Berlin Heidelberg 1999

Authors and Affiliations

  • N. Itoh
    • 1
  • Y. Tujibata
    • 2
  • J. Q. Liu
    • 1
  1. 1.Biotechnology 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: +81-766-56-2498JP
  2. 2.Department of Applied Chemistry and Biotechnology, Faculty of Engineering, Fukui University, Bunkyo 3-9-1, Fukui 910-8507, JapanJP