Expression of xylose isomerase activity in yeast
AssigneeE. I. Du Pont de Nemours and Company
InventorsWILLIAM D HITZ, Min Qi, Sarah Eve Rush, Luan Tao, Paul V. Viitanen, Jianjun Yang, Rick W. Ye
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Abstract
Expression of a xylose isomerase in a yeast cell that expresses the chaperonins GroES and GroEL was found to result in enzymatically active xylose isomerase, while there is little to no activity with expression of the bacterial xylose isomerase in a yeast cell lacking GroES and GroEL. A yeast cell expressing xylose isomerase activity, and a complete xylose utilization pathway, provides a yeast cell that can produce a target compound, such as ethanol, butanol, or 1,3-propanediol, using xylose derived from lignocellulosic biomass as a carbon source.
Claims19
A recombinant yeast cell comprising:
- a) at least one gene encoding each amino acid sequence of an interacting pair of Group I chaperonin polypeptides;
- and
- b) at least one gene encoding a bacterial xylose isomerase polypeptide;
- wherein:
- i) the interacting pair of Group I chaperonins are active in the cytosol of the cell;
- ii) the bacterial xylose isomerase polypeptide is converted to an active xylose isomerase enzyme;
- and
- iii) the specific activity of the bacterial xylose isomerase enzyme is higher as compared with the specific activity of the same xylose isomerase enzyme expressed in the absence of the interacting pair of Group I chaperonin polypeptides.
A method for producing a yeast strain that has xylose isomerase activity comprising:
- a) providing a yeast cell;
- b) introducing a heterologous nucleic acid molecule encoding a GroEL polypeptide and a heterologous nucleic acid molecule encoding a GroES polypeptide;
- and
- c) introducing a heterologous nucleic acid molecule encoding a bacterial xylose isomerase polypeptide;
- wherein:
- i) the GroEL and GroES polypeptides are expressed in the cytosol of the cell;
- ii) the xylose isomerase polypeptide is converted to an active xylose isomerase enzyme;
- and
- iii) the specific activity of the xylose isomerase enzyme is higher as compared with the specific activity of the same xylose isomerase enzyme expressed in the absence of the GroEL and GroES polypeptides.
Dependencies are inferred from the claim text.