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Removing the Active‐Site Flap in Lipase A from Candida antarctica Produces a Functional Enzyme without Interfacial Activation
Author(s) -
Wikmark Ylva,
Engelmark Cassimjee Karim,
Lihammar Richard,
Bäckvall JanE.
Publication year - 2016
Publication title -
chembiochem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.05
H-Index - 126
eISSN - 1439-7633
pISSN - 1439-4227
DOI - 10.1002/cbic.201500471
Subject(s) - candida antarctica , chemistry , lipase , hydrolysis , enzyme , active site , stereoselectivity , enzyme kinetics , stereochemistry , alcohol , solvent , wild type , organic chemistry , catalysis , biochemistry , mutant , gene
A mobile region is proposed to be a flap that covers the active site of Candida antarctica lipase A. Removal of the mobile region retains the functional properties of the enzyme. Interestingly interfacial activation, required for the wild‐type enzyme, was not observed for the truncated variant, although stability, activity, and stereoselectivity were very similar for the wild‐type and variant enzymes. The variant followed classical Michaelis–Menten kinetics, unlike the wild type. Both gave the same relative specificity in the transacylation of a primary and a secondary alcohol in organic solvent. Furthermore, both showed the same enantioselectivity in transacylation of alcohols and the hydrolysis of alcohol esters, as well as in the hydrolysis of esters chiral at the acid part.