1. Modification of fatty acid selectivity of Candida antarctica lipase A by error-prone PCR
- Author
-
Dagmara Głód
- Subjects
0301 basic medicine ,Conjugated linoleic acid ,Mutant ,Bioengineering ,Protein Engineering ,Polymerase Chain Reaction ,Applied Microbiology and Biotechnology ,Fungal Proteins ,03 medical and health sciences ,Hydrolysis ,chemistry.chemical_compound ,Selectivity ,Linoleic Acids, Conjugated ,Lipase ,Triglycerides ,Candida ,chemistry.chemical_classification ,Fungal protein ,030109 nutrition & dietetics ,Error-prone PCR ,biology ,Fatty acid ,General Medicine ,biology.organism_classification ,Original Research Paper ,030104 developmental biology ,chemistry ,Biochemistry ,biology.protein ,lipids (amino acids, peptides, and proteins) ,Candida antarctica ,Directed Molecular Evolution ,Biotechnology - Abstract
Objective To generate Candida antarctica lipase A (CAL-A) mutants with modified fatty acid selectivities and improved lipolytic activities using error-prone PCR (epPCR). Results A Candida antarctica lipase A mutant was obtained in three rounds of epPCR. This mutant showed a 14 times higher ability to hydrolyze triacylglycerols containing conjugated linoleic acids, and was 12 and 14 times more selective towards cis-9, trans-11 and trans-10, cis-12 isomers respectively, compared to native lipase. Lipolytic activities towards fatty acid esters were markedly improved, in particular towards butyric, lauric, stearic and palmitic esters. Conclusion Directed molecular evolution is an efficient method to generate lipases with desirable selectivity towards CLA isomers and improved lipolytic activities towards esters of fatty acids. Electronic supplementary material The online version of this article (doi:10.1007/s10529-017-2299-0) contains supplementary material, which is available to authorized users.
- Published
- 2017
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