Genetic engineering of Pichia stipitis for fermentation of xylose

Thomas W. Jeffries; N. Q. Shi; J. Y. Cho; P. Lu; K. Dahn; J. Hendrick; H. K. Sreenath

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Genetic engineering of Pichia stipitis for fermentation of xylose

Informally Refereed

Authors:

Thomas W. Jeffries, N. Q. Shi, J. Y. Cho, P. Lu, K. Dahn, J. Hendrick, H. K. Sreenath

Year:

1998

Type:

Miscellaneous Publication

Station:

Forest Products Laboratory

Source:

7th International Conference on Biotechnology in the Pulp and Paper Industry : Vancouver, BC, Canada, June 16-19, 1998. Poster presentations, Vol. C. Montreal, Quebec : Technical Section, CPPA, 1998.:p. C215-C218.

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Abstract

A useful genetic system has been developed for the transformation of Pichia stipitis. This includes two selectable markers (URA3 and LEU2), integrating and autonomous replication vectors, a pop-out cassette that enables multiple targeted disruptions, and a genomic X-library for rapid cloning. Using this system we have cloned two genes for alcohol dehydrogenase (PsADH1 and PsADH2). two genes for pyruvate decarboxylase (PsPDC1 and PsPDC2). and cytochrome c (PsCYC1). Disruption of PsADH1, but not PsADH2, resulted in increased xylitol production. Disruption of PsCYC1 resulted in lower growth and higher ethanol yields. We have used it to heterologously express the gene for S. cerevisiae dihydroorotate dehydrogenase. This conferred the capacity for anaerobic growth.

Keywords

Pichia stipitis, Xylose, Genetic engineering, Fermentation, Genetic markers, Genetic transformation, Cloning

Citation

Jeffries, Thomas W.; Shi, N. Q.; Cho, J. Y.; Lu, P.; Dahn, K.; Hendrick, J.; Sreenath, H. K. 1998. Genetic engineering of Pichia stipitis for fermentation of xylose. 7th International Conference on Biotechnology in the Pulp and Paper Industry : Vancouver, BC, Canada, June 16-19, 1998. Poster presentations, Vol. C. Montreal, Quebec : Technical Section, CPPA, 1998.:p. C215-C218.