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Protein engineering of the CalB lipase to synthesize methyl salicylate

Chew Fajardo, Ying Lian; Lopez de Los Santos, Yossef; Brault, Guillaume; Doucet, Nicolas . Protein engineering of the CalB lipase to synthesize methyl salicylate In: 2nd Protein Engineering Canada Conference (PEC), 17-19 Juin 2016, Ottawa, Canada.

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Résumé

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 flavoring 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], offering potential for the biological production of flavoring agents. However, the relatively confined organization of its active site precludes the recognition of more complex substrates. To overcome this limitation, in silico docking analyses of the best clones obtained from a previous mutant library generated in the Doucet lab will be undertaken. This will allow identification of the most significant amino acid residues involved in methyl salicylate precursor binding and recognition. 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 activity. Finally, up scaling production of the most efficient variants will be tested to help develop a biocatalyst for the proper industrial enzymatic synthesis of this flavor.

Type de document: Document issu d'une conférence ou d'un atelier
Mots-clés libres: -
Centre: Centre INRS-Institut Armand Frappier
Date de dépôt: 13 mars 2019 15:26
Dernière modification: 13 mars 2019 15:26
URI: http://espace.inrs.ca/id/eprint/5786

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