Disclosed in the present invention is a Zymomonas integrant 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 the Zymomonas in 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. The
Zymomonas integrants 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 unique Zymomonas that may then be used for the efficient
conversion of pentose sugars (xylose, arabinose) to ethanol. Method of
using the Zymomonas integrants and derivatives thereof in production of
ethanol from cellulosic feedstock is also disclosed. The invention also
provides a method for preparing a Zymomonas integrant as part of the
present invention. The host Zymomonas strain found particularly useful in
the creation of these compositions and methods is Zymomonas mobilis
31821.