Octopine dehydrogenase (ODH) catalyzes the final step of glucose fermentation in the bay scallop Argopecten irradians, producing octopine and NAD+ from pyruvate, arginine and NADH. Three stably maintained alleles of the ODH gene were identified in Argopecten populations in coastal areas of Long Island, on the basis of differential protein electrophoretic mobility. The alleles differ by one to three amino acid substitutions outside the active site. To determine the functional differences between them, the three alleles were subcloned into a bacterial expression vector, expressed in Escherichia coli as fusion proteins with an N-terminal affinity tag and purified by column chromatography. The affinity tag was removed by site-specific proteolysis and subsequent chromatography, yielding 10-15 mg of native enzyme per liter of culture. Preliminary data suggest that the three enzyme variants differ in their amino acid substrate preference.

Cloning, overexpression, and purification of octopine dehydrogenase from Argopecten irradians

245th National Meeting (2013)

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