Comparison of engineered Saccharomyces cerevisiae and engineered Escherichia coli for the production of an optically pure keto alcohol.
In this study, the production of enantiomerically pure (1R,4S,6S)-6-hydroxy-bicyclo[2.2.2]octane-2-one ((-)-2) through stereoselective bioreduction was used as a model reaction for the comparison of engineered Saccharomyces cerevisiae and engineered Escherichia coli as biocatalysts. For both microorganisms, over-expression of the gene encoding the NADPH-dependent aldo-keto reductase YPR1 resulted in high purity of the keto alcohol (-)-2 (>99% ee, 97-98% de). E. coli had three times higher initial reduction rate but S. cerevisiae continued the reduction reaction for a longer time period, thus reaching a higher conversion of the substrate (95%). S. cerevisiae was also more robust than E. coli, as demonstrated by higher viability during bioreduction. It was also investigated whether the NADPH regeneration rate was sufficient to supply the over-expressed reductase with NADPH. Five strains of each microorganism with varied carbon flux through the NADPH regenerating pentose phosphate pathway were genetically constructed and compared. S. cerevisiae required an increased NADPH regeneration rate to supply YPR1 with co-enzyme while the native NADPH regeneration rate was sufficient for E. coli.
Department of Applied Microbiology, Center for Chemistry and Chemical engineering, Lund University, P.O. Box 124, 221 00, Lund, Sweden.
Nádia
N
Skorupa Parachin
Magnus
M
Carlquist
Marie F
MF
Gorwa-Grauslund
D000429:Alcohol Oxidoreductases / Q000235:genetics / Q000378:metabolism; D000438:Alcohols / Q000378:metabolism*; D000449:Aldehyde Reductase; D000074408:Aldo-Keto Reductases; D001709:Biotechnology / Q000379:methods*; D001643:Bridged Bicyclo Compounds / Q000378:metabolism; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics / Q000254:growth & development / Q000378:metabolism; D007659:Ketones / Q000378:metabolism*; D009249:NADP / Q000378:metabolism; D030781:Organisms, Genetically Modified; D010084:Oxidation-Reduction; D012441:Saccharomyces cerevisiae / Q000201:enzymology / Q000235:genetics / Q000254:growth & development / Q000378:metabolism; D013237:Stereoisomerism
D000438:Alcohols; D001643:Bridged Bicyclo Compounds; D007659:Ketones; D009249:NADP; D000429:Alcohol Oxidoreductases; D000074408:Aldo-Keto Reductases; D000449:Aldehyde Reductase