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Fermentation 2018, 4(3), 59; https://doi.org/10.3390/fermentation4030059

Recombinant Diploid Saccharomyces cerevisiae Strain Development for Rapid Glucose and Xylose Co-Fermentation

School of Life Sciences and Chemical Technology, Ngee Ann Polytechnic, Singapore 599489, Singapore
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Received: 25 June 2018 / Revised: 23 July 2018 / Accepted: 25 July 2018 / Published: 30 July 2018
(This article belongs to the Special Issue Yeast Biotechnology 2.0)
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Abstract

Cost-effective production of cellulosic ethanol requires robust microorganisms for rapid co-fermentation of glucose and xylose. This study aims to develop a recombinant diploid xylose-fermenting Saccharomyces cerevisiae strain for efficient conversion of lignocellulosic biomass sugars to ethanol. Episomal plasmids harboring codon-optimized Piromyces sp. E2 xylose isomerase (PirXylA) and Orpinomyces sp. ukk1 xylose (OrpXylA) genes were constructed and transformed into S. cerevisiae. The strain harboring plasmids with tandem PirXylA was favorable for xylose utilization when xylose was used as the sole carbon source, while the strain harboring plasmids with tandem OrpXylA was beneficial for glucose and xylose cofermentation. PirXylA and OrpXylA genes were also individually integrated into the genome of yeast strains in multiple copies. Such integration was beneficial for xylose alcoholic fermentation. The respiration-deficient strain carrying episomal or integrated OrpXylA genes exhibited the best performance for glucose and xylose co-fermentation. This was partly attributed to the high expression levels and activities of xylose isomerase. Mating a respiration-efficient strain carrying the integrated PirXylA gene with a respiration-deficient strain harboring integrated OrpXylA generated a diploid recombinant xylose-fermenting yeast strain STXQ with enhanced cell growth and xylose fermentation. Co-fermentation of 162 g L−1 glucose and 95 g L−1 xylose generated 120.6 g L−1 ethanol in 23 h, with sugar conversion higher than 99%, ethanol yield of 0.47 g g−1, and ethanol productivity of 5.26 g L−1·h−1. View Full-Text
Keywords: Saccharomyces cerevisiae; diploid; xylose isomerase; xylose fermentation; glucose and xylose co-fermentation; biomass hydrolysate; cellulosic ethanol Saccharomyces cerevisiae; diploid; xylose isomerase; xylose fermentation; glucose and xylose co-fermentation; biomass hydrolysate; cellulosic ethanol
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited (CC BY 4.0).

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Liu, T.; Huang, S.; Geng, A. Recombinant Diploid Saccharomyces cerevisiae Strain Development for Rapid Glucose and Xylose Co-Fermentation. Fermentation 2018, 4, 59.

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