Cellulase variants with reduced inhibition by glucose

C - Chemistry – Metallurgy – 12 – N

Patent

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Details

C12N 9/42 (2006.01) C12N 1/15 (2006.01) C12N 1/19 (2006.01) C12N 15/56 (2006.01) C12N 15/80 (2006.01) C12P 19/02 (2006.01)

Patent

CA 2712151

A Family 6 cellulase variant enzyme comprising one or more than one amino acid substitution selected from a basic, polar or non-polar amino acid at position 103, a valine or isoleucine at position 136, a tyrosine at position 186, a glutamic acid or glutamine at position 365 and a glutamine at position 410 is provided (said position determined from alignment of the parental Family 6 with SEQ ID NO: 1). Genetic constructs and genetically modified microbes comprising DNA sequences encoding the Family 6 cellulase variant are also provided. Family 6 cellulases of the invention display reduced inhibition by glucose relative to the parent Family 6 cellulases. Such cellulases find use in a variety of applications in industry, e.g., in the hydrolysis of pretreated lignocellulosic feedstock, that require cellulose activity in the presence glucose concentrations that would otherwise inhibit the activity of the parental enzyme.

L'invention porte sur une enzyme variante de la cellulase de la Famille 6 comprenant une ou plusieurs substitutions d'acides aminés choisies parmi un acide aminé basique, polaire ou non polaire en position 103, une valine ou isoleucine en position 136, une tyrosine en position 186, un acide glutamique ou une glutamine en position 365 et une glutamine en position 410 (ladite position étant déterminée par alignement de la Famille parentale 6 avec SEQ ID NO : 1). L'invention porte également sur des produits de construction génétiques et des microbes modifiés génétiquement comprenant des séquences d'ADN codant pour le variant de cellulase de la Famille 6. Les cellulases de la Famille 6 de l'invention présentent une inhibition par le glucose réduite par rapport aux cellulases parentes de la Famille 6. De telles cellulases trouvent une utilisation dans une diversité d'applications industrielles telles que l'hydrolyse d'une charge d'alimentation lignocellulosique prétraitée, qui nécessitent une activité cellulase en présence de concentrations de glucose qui inhiberaient sinon l'activité de l'enzyme parentale.

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