화학공학소재연구정보센터
Applied Microbiology and Biotechnology, Vol.42, No.6, 916-922, 1995
Electromicrobial Regeneration of Pyridine-Nucleotides and Other Preparative Redox Transformations with Clostridium-Thermoaceticum
Clostridium thermoaceticum contains interesting enzymes suitable for redox reactions. Various AMAPOR (artificial-mediator-accepting pyridine-nucleotide oxidoreductase) activities were used for electromicrobial pyridine nucleotide regeneration. The combination of AMAPOR with commercially available pyridine-nucleotide-dependent oxidoreductases led to (S)-glutamate, (2R,3S)-isocitrate, (2S,3R)-isocitrate, 6-phosphogluconate and ribulose 5-phosphate. The redox equivalents were provided by electrochemically regenerated artificial mediators. Methylviologen or cobalt sepulchrate were used for NAD(P)H regeneration, whereas carboxamidomethylviologen (CAV) or anthraquinone sulphonates (AQ-S) were suitable for NAD(P)(+) regeneration. With resting cells of C. thermoaceticum productivity numbers {mmol product/[biocatalyst (kg dry weight) x time (h)]} of about 30000 for NADPH, 7000 for NADH and 14000 for NADP(+) regeneration could be reached. The cycle number for NADPH regeneration was up to 4300, that for NADP(+) regeneration was at least 1600. An aldehyde and an alcohol oxidoreductase were used to reduce non-activated carboxylic acids to the alcohols and to dehydrogenate primary alcohols to the aldehydes or carboxy lates. The electromicrobial reduction of 6-chloropyridine 3-carboxylate to the corresponding alcohol was compared with the reduction by CO as electron donor. The application of phenothiazine-dye-type mediators (thionine, methylene blue) converted primary alcohols to the aldehydes with productivity numbers up to 1400 in the presence of hydrazine as aldehyde scavenger. With CAV or AQ-S, alcohols were dehydrogenated to carboxylic acids with productivity numbers of almost 1700.