Cloning and overexpression in Escherichia coli of the gene encoding dihydroxyacetone kinase isoenzyme I from Schizosaccharomyces pombe, and its application to dihydroxyacetone phosphate production |
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| Authors: | N Itoh Y Tujibata J Q Liu |
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| Institution: | (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: +81-766-56-2498, JP;(2) Department of Applied Chemistry and Biotechnology, Faculty of Engineering, Fukui University, Bunkyo 3-9-1, Fukui 910-8507, Japan, JP |
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| 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.
Received: 25 August 1998 / Received revision: 22 September 1998 / Accepted: 11 October 1998 |
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