Mutations in the stereospecificity pocket and at the entrance of the active site of Candida antarctica lipase B enhancing enzyme enantioselectivity
2010 (English)In: Journal of Molecular Catalysis B: Enzymatic, ISSN 1381-1177, Vol. 65, no 1-4, 11-17 p.Article in journal (Refereed) Published
Two different parts of Candida antarctica lipase B (stereospecificity pocket at the bottom of the active site and hydrophobic tunnel leading to the active site) were redesigned by single- or double-point mutations, in order to better control and improve enzyme enantioselectivity toward secondary alcohols. Single-point isosteric mutations of Ser47 and Thr42 situated in the stereospecificity pocket gave rise to variants with doubled enantioselectivity toward pentan-2-ol, in solid/gas reactor. Besides, the width and shape of the hydrophobic tunnel leading to the active site was modified by producing the following single-point mutants: Ile189Ala, Leu278Val and Ala282Leu. For each of these variants a significant modification of enantioselectivity was observed compared to wild-type enzyme, indicating that discrimination of the enantiomers by the enzyme could also arise from their different accessibilities from the enzyme surface to the catalytic site. (C) 2010 Elsevier B.V. All rights reserved.
Place, publisher, year, edition, pages
2010. Vol. 65, no 1-4, 11-17 p.
Lipase B from Candida antarctica, Stereoselective catalysis, Protein engineering, Stereospecificity pocket, Substrate accessibility to the active site
Biochemistry and Molecular Biology Biochemistry and Molecular Biology Physical Chemistry
IdentifiersURN: urn:nbn:se:kth:diva-29689DOI: 10.1016/j.molcatb.2010.01.007ISI: 000278926300003ScopusID: 2-s2.0-77952583523OAI: oai:DiVA.org:kth-29689DiVA: diva2:398054
QC 201102162011-02-162011-02-112011-05-12Bibliographically approved