Improved synthesis of chiral alcohols with Escherichia coli cells co-expressing pyridine nucleotide transhydrogenase, NADP+-dependent alcohol dehydrogenase and NAD+-dependent formate dehydrogenase.
Recombinant pyridine nucleotide transhydrogenase (PNT) from Escherichia coli has been used to regenerate NAD+ and NADPH. The pnta and pntb genes encoding for the alpha- and beta-subunits were cloned and co-expressed with NADP+-dependent alcohol dehydrogenase (ADH) from Lactobacillus kefir and NAD+-dependent formate dehydrogenase (FDH) from Candida boidinii. Using this whole-cell biocatalyst, efficient conversion of prochiral ketones to chiral alcohols was achieved: 66% acetophenone was reduced to (R)-phenylethanol over 12 h, whereas only 19% (R)-phenylethanol was formed under the same conditions with cells containing ADH and FDH genes but without PNT genes. Cells that were permeabilized with toluene showed ketone reduction only if both cofactors were present.
Weckbecker
Andrea
A
Institut für Molekulare Enzymtechnologie der Heinrich-Heine-Universität, Forschungszentrum Jülich, Wilhelm-Johnen-Str., Jülich, D-52426, Germany.
Hummel
Werner
W
D000098:Acetophenones / Q000378:metabolism*; D000429:Alcohol Oxidoreductases / Q000235:genetics / Q000378:metabolism*; D002175:Candida / Q000201:enzymology / Q000235:genetics; D002384:Catalysis; D003067:Coenzymes / Q000235:genetics / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D005560:Formate Dehydrogenases / Q000235:genetics / Q000378:metabolism*; D024861:Genetic Enhancement / Q000379:methods; D007536:Isomerism; D007778:Lactobacillus / Q000201:enzymology / Q000235:genetics; D009250:NADP Transhydrogenases / Q000235:genetics / Q000378:metabolism*; D010626:Phenylethyl Alcohol / Q000378:metabolism*; D015202:Protein Engineering / Q000379:methods*; D011994:Recombinant Proteins / Q000378:metabolism
D000098:Acetophenones; D003067:Coenzymes; D011994:Recombinant Proteins; D000429:Alcohol Oxidoreductases; C084155:alcohol dehydrogenase (NAD(P)+); D005560:Formate Dehydrogenases; D009250:NADP Transhydrogenases; D010626:Phenylethyl Alcohol; C038699:acetophenone