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

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. Ep...

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Main Authors: Tingting Liu, Shuangcheng Huang, Anli Geng
Format: Article
Language:English
Published: MDPI AG 2018-07-01
Series:Fermentation
Subjects:
Online Access:http://www.mdpi.com/2311-5637/4/3/59
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author Tingting Liu
Shuangcheng Huang
Anli Geng
author_facet Tingting Liu
Shuangcheng Huang
Anli Geng
author_sort Tingting Liu
collection DOAJ
description 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.
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spelling doaj.art-0f8c15723d4d44b5b7a86e4db6ee69022022-12-22T01:16:37ZengMDPI AGFermentation2311-56372018-07-01435910.3390/fermentation4030059fermentation4030059Recombinant Diploid Saccharomyces cerevisiae Strain Development for Rapid Glucose and Xylose Co-FermentationTingting Liu0Shuangcheng Huang1Anli Geng2School of Life Sciences and Chemical Technology, Ngee Ann Polytechnic, Singapore 599489, SingaporeSchool of Life Sciences and Chemical Technology, Ngee Ann Polytechnic, Singapore 599489, SingaporeSchool of Life Sciences and Chemical Technology, Ngee Ann Polytechnic, Singapore 599489, SingaporeCost-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.http://www.mdpi.com/2311-5637/4/3/59Saccharomyces cerevisiaediploidxylose isomerasexylose fermentationglucose and xylose co-fermentationbiomass hydrolysatecellulosic ethanol
spellingShingle Tingting Liu
Shuangcheng Huang
Anli Geng
Recombinant Diploid Saccharomyces cerevisiae Strain Development for Rapid Glucose and Xylose Co-Fermentation
Fermentation
Saccharomyces cerevisiae
diploid
xylose isomerase
xylose fermentation
glucose and xylose co-fermentation
biomass hydrolysate
cellulosic ethanol
title Recombinant Diploid Saccharomyces cerevisiae Strain Development for Rapid Glucose and Xylose Co-Fermentation
title_full Recombinant Diploid Saccharomyces cerevisiae Strain Development for Rapid Glucose and Xylose Co-Fermentation
title_fullStr Recombinant Diploid Saccharomyces cerevisiae Strain Development for Rapid Glucose and Xylose Co-Fermentation
title_full_unstemmed Recombinant Diploid Saccharomyces cerevisiae Strain Development for Rapid Glucose and Xylose Co-Fermentation
title_short Recombinant Diploid Saccharomyces cerevisiae Strain Development for Rapid Glucose and Xylose Co-Fermentation
title_sort recombinant diploid saccharomyces cerevisiae strain development for rapid glucose and xylose co fermentation
topic Saccharomyces cerevisiae
diploid
xylose isomerase
xylose fermentation
glucose and xylose co-fermentation
biomass hydrolysate
cellulosic ethanol
url http://www.mdpi.com/2311-5637/4/3/59
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AT shuangchenghuang recombinantdiploidsaccharomycescerevisiaestraindevelopmentforrapidglucoseandxylosecofermentation
AT anligeng recombinantdiploidsaccharomycescerevisiaestraindevelopmentforrapidglucoseandxylosecofermentation