Protein engineering of the CalB lipase to synthesize methyl salicylate
A computationally-guided semi-rational protein design approach will be used to improve the enzymatic selectivity and catalytic efficiency of the lipase B from Pseudozyma antarctica (CalB) to synthesize methyl salicylate. This fatty acid ester is a fla voring and fragrance compound with significant r...
| Main Authors: | , , , |
|---|---|
| Format: | Other/Unknown Material |
| Language: | unknown |
| Subjects: | |
| Online Access: | https://espace.inrs.ca/id/eprint/5798/ http://www.proteo.ca/downloads/Symposium-PROTEO-2016.pdf |
| Summary: | A computationally-guided semi-rational protein design approach will be used to improve the enzymatic selectivity and catalytic efficiency of the lipase B from Pseudozyma antarctica (CalB) to synthesize methyl salicylate. This fatty acid ester is a fla voring and fragrance compound with significant relevance in the biotechnological industry. CalB is one the most widely used lipases for the enzymatic hydrolysis and synthesis of esters [1,2,3,4,5], o ffering potential for the biological production of flavoring agents. However, the relatively confine d organization of its active site precludes the recognition of more complex substrates. To overco me this limitation, in silico docking analyses of the best clones obtained from a previous mu tant library generated in the Doucet lab will be undertaken. This will allow identification o f the most significant amino acid residues involved in methyl salicylate precursor binding and re cognition. These “hot spots” will be subjected to combinatorial mutagenesis to synthesize a ‘second generation’ library of CalB variants, which will further be screened for the desired activi ty. Finally, up scaling production of the most efficient variants will be tested to help develop a bi ocatalyst for the proper industrial enzymatic synthesis of this flavor. |
|---|