Creation of a synthetically useful lipase with higher than wild-type enantioselectivity and maintained catalytic activity.
[reaction: see text] We have found that two Geotrichum candidum lipase isozymes have remarkably different abilities to differentiate between enantiomers of ethyl 2-methyldecanoate. By rational recombination of selected portions of the two isozymes, we have created a novel lipase with an enantioselectivity superior to that of the best wild-type parent isozyme. Site-directed mutagenesis identified two key amino acid residues responsible for the improved enantioselectivity without compromised total activity of the reengineered enzyme.
Holmquist
M
M
Department of Biotechnology, Royal Institute of Technology, SE-100 44 Stockholm, Sweden. mats.holmquist@biochem.kth.se
Berglund
P
P
D000596:Amino Acids / Q000737:chemistry; D002384:Catalysis; D003651:Decanoates / Q000737:chemistry; D005848:Geotrichum / Q000201:enzymology; D007527:Isoenzymes; D008049:Lipase / Q000138:chemical synthesis* / Q000737:chemistry*; D016297:Mutagenesis, Site-Directed; D013237:Stereoisomerism; D013379:Substrate Specificity
D000596:Amino Acids; D003651:Decanoates; D007527:Isoenzymes; C408991:ethyl-2-methyldecanoate; D008049:Lipase