Panke Sven, Held Martin, Wubbolts Marcel G, Witholt Bernard, Schmid Andreas
Institute of Biotechnology, Swiss Federal Institute of Technology, ETH-Hönggerberg HPT, 8093 Zurich, Switzerland.
Biotechnol Bioeng. 2002 Oct 5;80(1):33-41. doi: 10.1002/bit.10346.
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.
重组大肠杆菌JM101(pSPZ10)细胞可产生假单胞菌属菌株VLB120的苯乙烯单加氧酶，该酶能将苯乙烯氧化为对映体过量大于99%的(S)-环氧苯乙烷。在30升双液相补料分批生物转化中，这种生物催化剂用于生产388克环氧苯乙烷。在超过10小时的时间内，平均总体积活性为每升170单位，在相比为0.5时，相当于每升每小时10.2毫摩尔的传质速率。在此传质速率下，生物转化系统似乎受底物传质限制。该反应器估计功率输入约为5 W·L⁻¹，这接近商业运行装置通常获得的值。产物可通过分馏轻松纯化至纯度超过97%。该过程说明了在含有有毒底物和产物的双液相系统中进行重组全细胞生物转化的可行性。
