Journal of Microbiology

The Microbiological Society of Korea (한국미생물학회)
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Characterization of Recombinant Drosophila melanogaster Myo-inositol-l-phosphate Synthase Expressed in Escherichia coli

  • Park, Sang-Hee (Department of Food and Microbial Technology, Seoul Women's University) ;
  • Kim, Jong-Il (Department of Food and Microbial Technology, Seoul Women's University)
  • Published : 2004.03.01
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Abstract

Cloned myo-inositol-1-phosphate synthase (INOS) of Drosophila melanogaster was expressed in Escherichia coli, and purified using a His-affinity column. The purified INOS required NAD$\^$+/ for the conversion of glucose-6-phosphate to inositol-1-phosphate. The optimum pH for myo-inositol-1-phosphate synthase is 7.5, and the maximum activity was measured at 40$^{\circ}C$. The molecular weight of the native enzyme, as determined by gel filtration, was approximately M$\_$r/ 271,000${\pm}$15,000. A single subunit of approximately M$\_$r/ 62,000${\pm}$5,000 was detected upon SDS-polyacrylamide gel electrophoresis. The Michaelis ($K_{m}$) and dissociation constants for glucose-6-phosphate were 3.5 and 3.7 mM, whereas for the cofactor NAD$\^$+/ these were 0.42 and 0.4 mM, respectively.

Keywords

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