title: Continuous fermentation studies with xylos-utilizing recombinant Zymomonas mobilis.
abstract: This study examined the continuous cofermentation performance characteristics of a dilute-acid "prehydrolysate-adapted" recombinant Zymomonas 39676:pZB4L and builds on the pH-stat batch fermentations with this recombinant that we reported on last year. Substitution of yeast extract by 1% (w/v) corn steep liquor (CSL) (50% solids) and Mg (2 mM) did not alter the cofermentation performance. Using declared assumptions, the cost of using CSL and Mg was estimated to be 12.5 cents/gal of ethanol with a possibility of 50% cost reduction using fourfold less CSL with 0.1% diammonium phosphate. Because of competition for a common sugar transporter that exhibits a higher affinity for glucose, utilization of glucose was complete whereas xylose was always present in the chemostat effluent. The ethanol yield, based on sugar used, was 94% of theoretical maximum. Altering the sugar ratio of the synthetic dilute acid hardwood prehydrolysate did not appear to significantly change the pattern of xylose utilization. Using a criterion of 80% sugar utilization for determining the maximum dilution rate (Dmax), changing the composition of the feed from 4% xylose to 3%, and simultaneously increasing the glucose from 0.8 to 1.8% shifted Dmax from 0.07 to 0.08/h. With equal amounts of both sugars (2.5%), Dmax was 0.07/h. By comparison to a similar investigation with rec Zm CP4:pZB5 with a 4% equal mixture of xylose and glucose, we observed that at pH 5.0, the Dmax was 0.064/h and shifted to 0.084/h at pH 5.75. At a level of 0.4% (w/v) acetic acid in the CSL-based medium with 3% xylose and 1.8% glucose at pH 5.75, the Dmax for the adapted recombinant shifted from 0.08 to 0.048/h, and the corresponding maximum volumetric ethanol productivity decreased 45%, from 1.52 to 0.84 g/(L.h). Under these conditions of continuous culture, linear regression of a Pirt plot of the specific rate of sugar utilization vs D showed that 4 g/L of acetic acid did not affect the maximum growth yield (0.030 g dry cell mass/g sugar), but did increase the maintenance coefficient twofold, from 0.46 to 1.0 g of sugar/(g of cell.h).
authors.affiliation: Department of Biochemistry, University of Toronto, Ontario, Canada. hugh.lawford@utoronto.ca
authors.forename: H G
authors.initials: HG
authors.lastname: Lawford
authors.forename: J D
authors.initials: JD
authors.lastname: Rousseau
authors.forename: A
authors.initials: A
authors.lastname: Mohagheghi
authors.forename: J D
authors.initials: JD
authors.lastname: McMillan
mesh_terms: D001709:Biotechnology / Q000379:methods; D003470:Culture Media; D000431:Ethanol; D005285:Fermentation; D007700:Kinetics; D010957:Plasmids; D011995:Recombination, Genetic; D012441:Saccharomyces cerevisiae / Q000254:growth & development; D014994:Xylose / Q000378:metabolism*; D017355:Zymomonas / Q000235:genetics / Q000254:growth & development* / Q000378:metabolism
chemical_list: D003470:Culture Media; D000431:Ethanol; D014994:Xylose