Archer Daniels Midland Company

Lhing-Yew Li

Kelli J Trei

Beth Fatland-Bloom

Sean Jordan

Kelli Jean Trei

Disclosed herein is a newly discovered problem and solution for engineeringto ferment xylose to make ethanol utilizing xylose isomerase to convert xylose to xylulose for entry, via xylulokinase, into the pentose phosphate pathway. The problem is that when grown on a media containing xylose xylitol tends to accumulate in the cell despite the absence of xylose reductase activity in. Xylitol inhibits the activity of xylose isomerases. One solution described is to simultaneously express an exogenous xylitol dehydrogenase along with the exogenous xylose isomerase while optionally also overexpressing xylulokinase in the absence of expression of a xylose reductase. Another solution is a xylose isomerase fromwhich is less inhibited by xylitol than other xylose isomerases, exemplified byxylose isomerase. Expression of thexylose isomerase may be used alone, or in combination with expression of a xylitol dehydrogenase and optionally over expression of xylulokinase to improve ethanol production from xylose.

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The invention relates to production of lysine, and provides several isolated polynucleotide molecules useful for the production of L-lysine. One such polynucleotide encodes an aspartate kinase (ask), an aspartate-semialdehyde dehydrogenase (asd) and a dihydrodipicolinate reductase (dapB). Other polypeptides encode ask, asd, dapB and a diaminopimelate dehydrogenase (ddh); ask, asd, dapB, ddh and an ORF2 poypeptide; and ask, asd dapB, ddh, ORF2 and a diaminopimelate decarboxylase (lysA). The invention further provides methods of making and using the polynucleotides, and methods to increase the production of L-lysine.

Polynucleotide constructs for increased lysine production

The invention relates to production of lysine, and provides several isolated polynucleotide molecules useful for the production of L-lysine. One such polynucleotide encodes an aspartate kinase (ask), an aspartate-semialdehyde dehydrogenase (asd) and a dihydrodipicolinate reductase. Other polypeptides encode ask, asd, dihydrodipicolinate reductase, and a diaminopimelate dehydrogenase (ddh); ask, asd, dihydrodipicolinate reductase, ddh, and an ORF2 polypeptide; and ask, asd, dihydrodipicolinate reductase, ddh, ORF2 and a diaminopimelate decarboxylase. The invention further provides methods of making and using the polynucleotides, and methods to increase the production of L-lysine. The invention further provides use of isolated polynucleotide molecules encoded by genes native to bacteria of the genus. The invention further provides host cells bearing the isolated polynucleotide molecules of the invention. The invention further provides embodiments in which the host cell is a member of the genus

Polynucleotide constructs encoding aspartate kinase, aspartate-semialdehyde dehydrogenase, and dihydrodipicolinate reductase and related constructs, products, and methods

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Savoy, IL, United States of America

Lhing-Yew Li