Fermentative production of isobutanol with yeast

Details Fermentative production of isobutanol with yeast Fermentative production of isobutanol with yeast

C - Chemistry, Metallurgy

12

P

C12P 7/16 (2006.01) C12N 9/04 (2006.01) C12N 9/10 (2006.01) C12N 9/88 (2006.01)

Patent

CA 2716427

The invention relates to a yeast cell producing isobutanol, characterized in that the cell has an increased metabolic flow of material from pyruvate and acetolactate, 2, 3-dihydroxy isovalerate, 2- ketoisovalerate, isobutyraldehyde to isobutanol, in that at least one of the genes coding the enzymes, which are involved in this conversion, is over-expressed, and without any of said genes being heterologous to said yeast cell, wherein Ilv2 (=YMR108W) (SEQ. ID. No. 2) catalyzes the acetolactate synthesis reaction from pyruvate to acetolactate, Ilv5 (=YLR355C) (SEQ. ID. No. 6) catalyzes the acetohydroxy acid reducto-isomerase reaction from acetolactate to 2, 3-dihydroxy isovalerate, Ilv3 (=YJR106C) (SEQ. ID. No. 8) catalyzes the dihydroxy acid dehydratase reaction from 2,3-dihydroxy isovalerate to 2-ketoisovalerate, a 2-keto acid decarboxylase catalyzes the reaction from 2-ketoisovalerate to isobutyraldehyde, and an alcohol dehydrogenase catalyzes the reaction from isobutyraldehyde to isobutanol, and to a method for the production of isobutanol using yeast cells, comprising the provision of the yeast cells according to the invention, and bringing the yeast cell into contact with a fermentable carbon source.

L'invention concerne une cellule de levure produisant de l'isobutanol, caractérisée en ce que la cellule présente un flux métabolique accru du pyruvate vers l'isobutanol par l'acétolactate, le 2,3-dihydroxy-isovalérate, le 2-céto-isovalérate, l'isobutyraldéhyde, par le fait qu'au moins un des gènes, codant pour l'enzyme participant à cette réaction, est surexprimé et sans qu'un de ces gènes ne soit hétérologue par rapport à ladite cellule de levure. Selon l'invention, Ilv2 (=YMR108W) (SEQ. ID. Nr. 2) catalyse la réaction de l'acétolactate synthase du pyruvate en acétolactate, Ilv5 (=YLR355C) (SEQ. ID. Nr. 6) catalyse la réaction de l'acétohydroxyacide réductoisomérase de l'acétolactate en 2, 3-dihydroxyisovalérate, Ilv3 (=YJR016C) (SEQ. ID. Nr. 8) cataylse la réaction de la dihydroxyacide déshydratase du 2,3-dihydroxyisovalérate en 2-céto-isovalérate, une 2- céto-acide décarboxylase catalyse la réaction du 2-céto-isovalérate en isobutyraldéhyde et une alcool déshydrogénase catalyse la réaction de l'isobutyraldéhyde en isobutanol. L'invention concerne également un procédé pour la production d'isobutanol au moyen de cellules de levure, comprenant la préparation des cellules de levure selon l'invention ainsi que la mise en contact de la cellule de levure avec une source de carbone fermentable.

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