✦ LIBER ✦

Pilot scale production and isolation of recombinant NAD+- and NADP+-specific formate dehydrogenases

✍ Scribed by Vladimir I. Tishkov; Andrey G. Galkin; Vladimir V. Fedorchuk; Pavel A. Savitsky; Alexandra M. Rojkova; Holger Gieren; Maria-Regina Kula

Publisher: John Wiley and Sons
Year: 1999
Tongue: English
Weight: 350 KB
Volume: 64
Category: Article
ISSN: 0006-3592
DOI: 10.1002/(sici)1097-0290(19990720)64:2<187::aid-bit7>3.0.co;2-0

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✦ Synopsis

The expression of the recombinant wild-type NAD + -and mutant NADP + -dependent formate dehydrogenases (EC 1.2.1.2., FDH) from the methanol-utilizing bacterium Pseudomonas sp. 101 in Escherichia coli cells has been improved to produce active and soluble enzyme up to the level of 50% of total soluble proteins. The cultivation process for E. coli/pFDH8a and E. coli/ pFDH8aNP cells was optimized and scaled up to a volume of 100 L. A downstream purification process has been developed to produce technical grade NAD + -and NADP + -specific formate dehydrogenases in pilot scale, utilizing extraction in aqueous two-phase systems.

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