Advanced Synthesis and Catalysis p. 126 - 134 (2019)
Update date:2022-08-03
Cen, Yixin
Li, Danyang
Xu, Jian
Wu, Qiongsi
Wu, Qi
Lin, Xianfu
Candida antarctica lipase B (CALB) is one of the most extensively used biocatalysts in both academia and industry and exhibits remarkable (R)-enantioselectivity for various chiral sec-alcohols. Considering the significance of tailor-made stereoselectivity in organic synthesis, a discovery of enantiocomplementary lipase mutants with high (R)- and (S)-selectivity is valuable and highly desired. Herein, we report a highly efficient directed evolution strategy, using only 4 representative amino acids, namely, alanine (A), leucine (L), lysine (K), tryptophan (W) at each mutated site to create an extremely small library of CALB variants requiring notably less screening. The obtained best mutant with three mutations W104V/A281L/A282K displayed highly reversed (S)-selectivity towards a series of sec-alcohol with E values up to 115 (conv. 50%, ee 94%). Compared with the previously reported (S)-selective CALB variant, W104A, a single mutation provided less selectivity, while the synergistic effects of three mutations in the best variant endow better (S)-selectivity and a broader substrate scope than the W104A variant. Structural analysis and molecular dynamics simulation unveiled the source of reversed enantioselectivity. (Figure presented.).
View MoreShanghai Yuanye Bio-Technology Co., Ltd.
website:http://www.shyuanye.com
Contact:+86-21-61845781
Address:Building 6, No. 465, Changta Road,Songjiang District,Shanghai,China
SuZhou Hua-Emy Chemical Import and Export Co., LTD.
Contact:+86-512-88804994; +86-512-88804550;
Address:710, Building B, International Trade Center, 12 Huanghelu, Changshu, Jiangsu,China
Chemsigma International Co.,Ltd.
website:http://www.chemsigma.com
Contact:86-025-58748998
Address:Rm.705, 15th Building,Rd.Xinke II
WUHU HUAHAI BIOLOGY ENGINEERING CO LTD
Contact:+86-553-3836920
Address:7/F NO.82 LAODONG ROAD WUHU CHINA
Contact:+86-371-55981030
Address:Room 1571, Macalline Soho, No.1, Shangdu Road, Zhengzhou, Henan
Doi:10.1021/om1002045
(2010)Doi:10.1039/c2md20332d
(2013)Doi:10.1002/ejoc.201000158
(2010)Doi:10.1016/j.bmc.2010.06.003
(2010)Doi:10.1016/j.bmc.2010.06.002
(2010)Doi:10.1134/S1070363210050117
(2010)