Reactor operation and scale-up of whole cell Baeyer-Villiger catalyzed lactone synthesis

被引:73
作者
Doig, SD [1 ]
Avenell, PJ [1 ]
Bird, PA [1 ]
Gallati, P [1 ]
Lander, KS [1 ]
Lye, GJ [1 ]
Wohlgemuth, R [1 ]
Woodley, JM [1 ]
机构
[1] UCL, Dept Biochem Engn, London WC1E 7JE, England
关键词
D O I
10.1021/bp0200954
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
The recombinant whole cell biocatalyst Escherichia coli TOP10 [pQR239], expressing cyclohexanone monooxygenase from Acinetobacter calcoaceticus NCIMB 987 1, was used in 1.5- and 55-L fed-batch processes to oxidize bicyclo[3.2.0]hept-2-en-6-one to its corresponding regioisomeric lactones, (-)-(1S,5R)-2-oxabicyclo[3.3.0]oct-6-en-3-one and (-)-(1R,5S)-3-oxabicyclo[3.3.0]oct-6-en-2-one. By employing a bicyclo[3.2.0]hept-2-en-6-one feed rate below that of the theoretical volumetric biocatalyst activity (275mumol(.)min(-1.)L(-1)), the reactant concentration in the bioreactor was successfully maintained below the inhibitory concentration of 0.2-0.4 g(.)L(-1). In this way approximately 3.5 g(.)L(-1) of the combined regioisomeric lactones was produced with a yield of product on reactant of 85-90%. The key limitation to the process was shown to be product inhibition. This process was scaled up to 55 L, producing over 200 g of combined lactone product. Using a simple downstream process (centrifugation, adsorption to-activated charcoal, 5-fold concentration with ethyl acetate elution, and silica gel chromatography), we have shown that the two regioisomeric lactone products could be isolated and purified at this scale.
引用
收藏
页码:1039 / 1046
页数:8
相关论文
共 25 条
[21]   THE ENZYMATIC BAEYER-VILLIGER OXIDATION - A STUDY OF 4-SUBSTITUTED CYCLOHEXANONES [J].
TASCHNER, MJ ;
BLACK, DJ ;
CHEN, QZ .
TETRAHEDRON-ASYMMETRY, 1993, 4 (06) :1387-1390
[22]  
TRUDGILL PW, 1990, METHOD ENZYMOL, V188, P70
[23]  
WILLETS AJ, 1991, J CHEM SOC P1, V6, P1608
[24]   Structural studies and synthetic applications of Baeyer-Villiger monooxygenases [J].
Willetts, A .
TRENDS IN BIOTECHNOLOGY, 1997, 15 (02) :55-62
[25]   Effect of substrate concentration on the enantioselectivity of cyclohexanone monooxygenase from Acinetobacter calcoaceticus and its rationalization [J].
Zambianchi, F ;
Pasta, P ;
Ottolina, G ;
Carrea, G ;
Colonna, S ;
Gaggero, N ;
Ward, JM .
TETRAHEDRON-ASYMMETRY, 2000, 11 (18) :3653-3657