Midwest Research Institute

Christine Eddy

Min Zhang

Yat-Chen Chou

Kent Evans

Ali Mohagheghi

William Howe

Disclosed in the present invention is aintegrant and derivatives of these integrants that posses the ability to ferment pentose into ethanol. The genetic sequences encoding for the pentose-fermenting enzymes are integrated into thein a two-integration event of homologous recombination and transposition. Each operon includes more than one pentose-reducing enzyme encoding sequence. The integrant in some embodiments includes enzyme sequences encoding xylose isomerase, xylulokinase, transketolase and transketolase. Theintegrants are highly stable, and retain activity for producing the pentose-fermenting enzyme for between 80 to 160 generations. The integrants are also resistant to acetate inhibition, as the integrants demonstrate efficient ethanol production even in the presence of 8 up to 16 grams acetate per liter media. These stably integrated sequences provide a uniquethat may then be used for the efficient conversion of pentose sugars (xylose, arabinose) to ethanol. Method of using theintegrants and derivatives thereof in production of ethanol from cellulosic feedstock is also disclosed. The invention also provides a method for preparing aintegrant as part of the present invention. The hoststrain found particularly useful in the creation of these compositions and methods is31821.

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Golden, CO, United States of America

Christine Eddy