A self-sufficient Baeyer-Villiger biocatalysis system for the synthesis of ɛ-caprolactone from cyclohexanol.
In order to establish a new route for ɛ-caprolactone production from the corresponding cyclohexanol with an internal cofactor recycling for NADPH, a recently redesigned thermostable polyol dehydrogenase (PDH) and the cyclohexanone monooxygenase (CHMO) from Acinetobacter calcoaceticus were combined. First, the expression of PDH could be improved 4.9-fold using E. coli C41 with co-expression of chaperones. Both enzymes were also successfully co-immobilized on glutaraldehyde-activated support (Relizyme™ HA403). Cyclohexanol could be converted to ɛ-caprolactone (ɛ-CL) with 83% conversion using the free enzymes and with 34% conversion using the co-immobilized catalysts. Additionally, a preparative scale biotransformation of ɛ-caprolactone starting from cyclohexanol was performed using the soluble enzymes. The ɛ-CL could be isolated by simple extraction and evaporation with a yield of 55% and a purity of >99%.
Department of Biotechnology & Enzyme Catalysis, Institute of Biochemistry, Greifswald University, Germany.
H
H
Mallin
H
H
Wulf
U T
UT
Bornscheuer
D016954:Acinetobacter calcoaceticus / Q000201:enzymology / Q000235:genetics; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D055162:Biocatalysis; D001709:Biotechnology; D001711:Biotransformation; D002208:Caproates / Q000378:metabolism*; D003511:Cyclohexanols / Q000378:metabolism*; D004800:Enzymes, Immobilized / Q000235:genetics / Q000378:metabolism; D007700:Kinetics; D007064:L-Iditol 2-Dehydrogenase / Q000235:genetics / Q000378:metabolism; D007783:Lactones / Q000378:metabolism*; D009249:NADP / Q000378:metabolism; D010105:Oxygenases / Q000235:genetics / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism
D001426:Bacterial Proteins; D002208:Caproates; D003511:Cyclohexanols; D004800:Enzymes, Immobilized; D007783:Lactones; D011993:Recombinant Fusion Proteins; D009249:NADP; C121056:caprolactone; D007064:L-Iditol 2-Dehydrogenase; D010105:Oxygenases; C019961:cyclohexanone oxygenase
BVMO; Baeyer–Villiger monooxygenase; Biocatalysis; CHL; CHMO; CHO; Co-immobilization; Cofactor recycling; FID; GA; PDH; cyclohexanol; cyclohexanone; cyclohexanone monooxygenase from Acinetobacter calcoaceticus; flame-ionization detection; glutaraldehyde; polyol dehydrogenase (PDH_loopN); ɛ-CL; ɛ-Caprolactone; ɛ-caprolactone