Improved Candida methylica formate dehydrogenase fermentation through statistical optimization of low-cost culture media.

NAD⁺-dependent formate dehydrogenase (FDH, EC 1.2.1.

Improving the purification of NAD+-dependent formate dehydrogenase from Candida methylica.

The Candida methylica (cm) recombinant wild type formate dehydrogenase (FDH) gene has been cloned into the pQE-2 TAGZyme expression vector and the 6xHis-tagged FDH gene has been overexpressed in JM105 cells to purify the FDH protein more efficiently, by the use of exopeptidases, TAGZyme Purification System, which has allowed the complete removal of the small N-terminal His-tag. After the purification procedure, 1.2 mg/mL cmFDH protein of >95% purity was obtained.