Family 6 cellulase with decreased inactivation by lignin

C - Chemistry – Metallurgy – 12 – N

Patent

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Details

C12N 9/42 (2006.01) C12N 15/56 (2006.01) C12N 15/80 (2006.01) C12P 19/00 (2006.01) C12P 21/02 (2006.01) C12S 3/04 (2006.01)

Patent

CA 2732082

A modified Trichoderma reesei Family 6 (TrCel6A) cellulase enzyme comprising amino acid substitutions at one or more positions selected from the group consisting of 129, 322, 363 and 410 of SEQ ID NO: 1 is provided. Genetic constructs and genetically modified microbes comprising nucleic sequences encoding the modified TrCelóa cellulase are also provided. The modified TrCel6A cellulase of the invention display at least a 15% decrease in inactivation by lignin relative to a parental TrCelóA cellulase from which the modified TrCel6A is derived. Such cellulases find use in a variety of applications in industry requiring enzymatic hydrolysis of cellulose in the presence of lignin, e.g., the hydrolysis of pretreated lignocellulosic feedstocks for the production of fermentable sugars, sugar alcohols and fuel alcohols.

L'invention porte sur une enzyme cellulase de la famille 6 de Trichoderma reesei (TrCel6A) modifiée renfermant des substitutions d'acides aminés au niveau d'une ou plusieurs positions choisies dans le groupe constitué par les positions 129, 322, 363 ou 410 de SEQ ID NO: 1. L'invention porte également sur des constructions génétiques et des microbes génétiquement modifiés renfermant des séquences nucléiques codant pour la cellulase TrCel6A modifiée. La cellulase TrCel6A modifiée de l'invention présente une diminution d'au moins 15 % de l'inactivation par la lignine par rapport à une cellulase TrCel6A parente dont la TrCel6A modifiée est issue. De telles cellulases trouvent une utilisation dans diverses applications dans l'industrie nécessitant une hydrolyse enzymatique de cellulose en présence de lignine, par exemple l'hydrolyse de charges d'alimentation lignocellulosiques prétraitées pour la production de sucres fermentescibles, d'alcools de sucre et d'alcools combustibles.

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