Journal of Microbiology

Research Support, Non-U.S. Gov't

Characterization of Recombinant Drosophila melanogaster Myo-inositol-1-phosphate Synthase Expressed in Escherichia coli: Sang-Hee Park , Jong-Il Kim; J. Microbiol. 2004;42(1):20-24.; DOI: https://doi.org/2006 [pii]

Abstract: Cloned myo-inositol-1-phpsphate 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^oC. The molecular weight of the native enzyme, as determined by gel filtration, was approximately M_r 271,000±15,000. A single subunit of approximately M_r 62,000±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.

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