Fermentation performance characteristics of a prehydrolyzate-adapted xylose-fermenting recombinant Zymomonas in batch and continuous fermentations |
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Authors: | Hugh G Lawford Joyce D Rousseau Ali Mohagheghi James D McMillan |
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Institution: | (1) Bioengineering Laboratory, Department of Biochemistry, University of Toronto, M5S 1A8 Toronto, Ontario, Canada;(2) Biotechnology Center for Fuels and Chemicals, National Renewable Energy Laboratory, Golden, CO |
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Abstract: | Long-term (149 d) continuous fermentation was used to adapt a xylose-fermenting recombinant Zymomonas mobilis, strain 39676:pZB 4L, to conditioned (overlimed) dilute-acid yellow poplar hemicellulose hydrolyzate (“prehydrolyzate”).
An “adapted” variant was isolated from a chemostat operating at a dilution rate of 0.03/h with a 50% (v/v) prehydrolyzate,
corn steep liquor, and sugar-supplemented medium, at pH 5.75. The level of xylose and glucose in the medium was kept constant
at 4% (w/v) and 0.8% (w/v), respectively. These sugar concentrations reflect the composition of the undiluted hardwood prehydrolyzate.
The level of conditioned hardwood prehydrolyzate added to the medium was increased in 5% increments startingata level of 10%.
At the upper level of 50% prehydrolyzate, the acetic-acid concentration was about 0.75% (w/v). The adapted variant exhibited
improved xylose-fermentation performance in a pure-sugar, synthetic hardwood prehydrolyzate medium containing 4% xylose (w/v),
0.8% (w/v) glucose, and acetic acid in the range 0.4–1.0% (w/v). The ethanol yield was 0.48–0.50 g/g; equivalent to a sugar-to-ethanol
conversion efficiency of 94–96% of theoretical maximum. The maximum growth yield and maintenance energy coefficients were
0.033 g dry cell mass (DCM)/g sugars and 0.41 g sugars/g DCM/h, respectively. The results confirm that long-term continuous
adaptation is a useful technique for effecting strain improvement with respect to the fermentation of recalcitrant feedstocks. |
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Keywords: | Recombinant Zymomonas continuous cofermentation xylose hardwood prehydrolyzate ethanol yield adaptation acetic acid |
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