An efficient bioprocess for enzymatic production of L-menthol with high ratio of substrate to catalyst using whole cells of recombinant E. coli.
A gene encoding an esterase of Bacillus subtilis ECU0554 previously isolated from soil was cloned and overexpressed in Escherichia coli BL21. The recombinant esterase (recBsE) showed the best enantioselectivity (E>100) towards DL-menthyl acetate, in contrast to DL-menthyl esters propionate and butyrate. A high ratio of substrate to catalyst (S/C-ratio, ≥50) was achieved in the kinetic resolution of DL-menthyl acetate by using whole cells of recombinant E. coli BL21. Some key parameters of the biocatalytic process, including amount of cosolvent, catalyst loading and substrate loading, were optimized. Compared with the process catalyzed by wild-type whole cells of B. subtilis ECU0554, the second-generation bioprocess using whole cells of recombinant E. coli BL21 afforded a 40-fold improvement in S/C-ratio and a 75-fold improvement in the volumetric productivity per biocatalyst loading. Moreover, the substrate loading was increased up to 200 g L(-1) (∼1 M), the biocatalyst loading was reduced to 2.5 g L(-1) and the space-time yield was improved from 54 g L(-1) d(-1) to 202 g L(-1) d(-1).
Zheng
Gao-Wei
GW
Laboratory of Biocatalysis and Bioprocessing, East China University of Science and Technology, 130 Meilong Road, Shanghai 200237, PR China.
Pan
Jiang
J
Yu
Hui-Lei
HL
Ngo-Thi
Minh-Thu
MT
Li
Chun-Xiu
CX
Xu
Jian-He
JH
D000085:Acetates / Q000378:metabolism; D001412:Bacillus subtilis / Q000201:enzymology / Q000235:genetics; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D019149:Bioreactors; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D004950:Esterases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D000431:Ethanol / Q000378:metabolism; D007700:Kinetics; D008610:Menthol / Q000032:analysis / Q000737:chemistry / Q000378:metabolism*; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
D000085:Acetates; D001426:Bacterial Proteins; D011994:Recombinant Proteins; D008610:Menthol; D000431:Ethanol; D004950:Esterases