Pilot-scale production of (S)-styrene oxide from styrene by recombinant Escherichia coli synthesizing styrene monooxygenase.
Recombinant Escherichia coli JM101(pSPZ10) cells produce the styrene monooxygenase of Pseudomonas sp. strain VLB120, which catalyzes the oxidation of styrene to (S)-styrene oxide at an enantiomeric excess larger than 99%. This biocatalyst was used to produce 388 g of styrene oxide in a two-liquid phase 30-L fed-batch bioconversion. The average overall volumetric activity was 170 U per liter over a period of more than 10 h, equivalent to mass transfer rates of 10.2 mmoles per liter per hour at a phase ratio of 0.5. At this transfer rate, the biotransformation system appeared to be substrate mass-transfer limited. The reactor had an estimated power input in the order of 5 W. L(-1), which is close to values typically obtained with commercially operating units. The product could be easily purified by fractional distillation to a purity in excess of 97%. The process illustrates the feasibility of recombinant whole cell biotransformations in two-liquid phase systems with toxic substrates and products.
Panke
Sven
S
Institute of Biotechnology, Swiss Federal Institute of Technology, ETH-Hönggerberg HPT, 8093 Zurich, Switzerland.
Held
Martin
M
Wubbolts
Marcel G
MG
Witholt
Bernard
B
Schmid
Andreas
A
D002460:Cell Line; D004852:Epoxy Compounds / Q000138:chemical synthesis* / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism*; D015964:Gene Expression Regulation, Bacterial; D010105:Oxygenases / Q000235:genetics / Q000378:metabolism*; D010865:Pilot Projects; D011549:Pseudomonas / Q000201:enzymology / Q000235:genetics; D011995:Recombination, Genetic; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013237:Stereoisomerism; D020058:Styrene / Q000378:metabolism*
D004852:Epoxy Compounds; D020058:Styrene; C013690:styrene oxide; D010105:Oxygenases; C026721:styrene monooxygenase