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Pilot scale production and isolation of recombinant NAD + ‐ and NADP + ‐specific formate dehydrogenases
Author(s) -
Tishkov Vladimir I.,
Galkin Andrey G.,
Fedorchuk Vladimir V.,
Savitsky Pavel A.,
Rojkova Alexandra M.,
Gieren Holger,
Kula MariaRegina
Publication year - 1999
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/(sici)1097-0290(19990720)64:2<187::aid-bit7>3.0.co;2-0
Subject(s) - formate dehydrogenase , nad+ kinase , formate , escherichia coli , recombinant dna , cofactor , biochemistry , enzyme , chemistry , mutant , methanol , chromatography , biology , organic chemistry , catalysis , gene
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. © 1999 John Wiley & Sons, Inc. Biotechnol Bioeng 64: 187–193, 1999.