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Rational investigation of the potential of Aspergillus strains for lipases production: from gene prospection to application in enantioselectivity models

Abstract

Different rational engineering approaches based on enzyme diversity and selecting variants with improved properties have been used to improve enzyme characteristics. The use of lipases is increasingly gaining focus due to the versatility of the catalyzed reactions and their regio, chemo and enantioselectivity. Expanding the knowledge of the enantioselectivity of lipases is of great interest considering it is (1) of great industrial relevance, (2) relatively easy to investigate, and (3) closely related to the structural arrangement of the active site of the enzyme, so that it preferentially recognizes one of the enantiomers, which enables the separation and generation of the isomer of interest. In this project, we propose an unprecedented study based on the rational engineering of Aspergillus niger, a lipase-producing species previously studied by the group, which should contribute to the research of new enzymes with high enantioselectivity. The Aspergillus genome harbors many unexplored enzyme genes, potentially hiding new lipases to be used in the market. The tools used involve gene bioprospection and the production of Aspergillus recombinant lipases using the CRISPR/Cas9 (Clustered Regularly Interspaced Short Palindromic Repeats) technique, purification and immobilization of the new enzymes and the subsequent application in enantioselectivity models involving hydrolysis, esterification and aminolysis reactions of intermediates and chiral drugs. We expect that the present project will expand the knowledge and the selection of new enzymes with improved characteristics as potential biocatalysts for the fine chemical industry. (AU)

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