scholarly journals Coutilization of D-Glucose, D-Xylose, and L-Arabinose in Saccharomyces cerevisiae by Coexpressing the Metabolic Pathways and Evolutionary Engineering

2017 ◽  
Vol 2017 ◽  
pp. 1-8 ◽  
Author(s):  
Chengqiang Wang ◽  
Jianzhi Zhao ◽  
Chenxi Qiu ◽  
Shihao Wang ◽  
Yu Shen ◽  
...  
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Ethanol Production ◽  
Metabolic Pathways ◽  
Ethanol Yield ◽  
Cost Effective ◽  
Evolutionary Engineering ◽  
Lignocellulosic Materials ◽  
Metabolic Capacity ◽  
Mixed Sugars ◽  
Consumption Rates

Efficient and cost-effective fuel ethanol production from lignocellulosic materials requires simultaneous cofermentation of all hydrolyzed sugars, mainly including D-glucose, D-xylose, and L-arabinose. Saccharomyces cerevisiae is a traditional D-glucose fermenting strain and could utilize D-xylose and L-arabinose after introducing the initial metabolic pathways. The efficiency and simultaneous coutilization of the two pentoses and D-glucose for ethanol production in S. cerevisiae still need to be optimized. Previously, we constructed an L-arabinose-utilizing S. cerevisiae BSW3AP. In this study, we further introduced the XI and XR-XDH metabolic pathways of D-xylose into BSW3AP to obtain D-glucose, D-xylose, and L-arabinose cofermenting strain. Benefits of evolutionary engineering: the resulting strain BSW4XA3 displayed a simultaneous coutilization of D-xylose and L-arabinose with similar consumption rates, and the D-glucose metabolic capacity was not decreased. After 120 h of fermentation on mixed D-glucose, D-xylose, and L-arabinose, BSW4XA3 consumed 24% more amounts of pentoses and the ethanol yield of mixed sugars was increased by 30% than that of BSW3AP. The resulting strain BSW4XA3 was a useful chassis for further enhancing the coutilization efficiency of mixed sugars for bioethanol production.

scholarly journals Directed Evolution of Xylose Isomerase for Improved Xylose Catabolism and Fermentation in the Yeast Saccharomyces cerevisiae

2012 ◽  
Vol 78 (16) ◽  
pp. 5708-5716 ◽  
Author(s):  
Sun-Mi Lee ◽  
Taylor Jellison ◽  
Hal S. Alper
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Ethanol Production ◽  
Directed Evolution ◽  
Ethanol Yield ◽  
Xylose Isomerase ◽  
Mutant Enzyme ◽  
Xylose Utilization ◽  
Xylose Consumption ◽  
Content Type ◽  
Consumption Rates

ABSTRACTThe heterologous expression of a highly functional xylose isomerase pathway inSaccharomyces cerevisiaewould have significant advantages for ethanol yield, since the pathway bypasses cofactor requirements found in the traditionally used oxidoreductase pathways. However, nearly all reported xylose isomerase-based pathways inS. cerevisiaesuffer from poor ethanol productivity, low xylose consumption rates, and poor cell growth compared with an oxidoreductase pathway and, additionally, often require adaptive strain evolution. Here, we report on the directed evolution of thePiromycessp. xylose isomerase (encoded byxylA) for use in yeast. After three rounds of mutagenesis and growth-based screening, we isolated a variant containing six mutations (E15D, E114G, E129D, T142S, A177T, and V433I) that exhibited a 77% increase in enzymatic activity. When expressed in a minimally engineered yeast host containing agre3knockout andtal1andXKS1overexpression, the strain expressing this mutant enzyme improved its aerobic growth rate by 61-fold and both ethanol production and xylose consumption rates by nearly 8-fold. Moreover, the mutant enzyme enabled ethanol production by these yeasts under oxygen-limited fermentation conditions, unlike the wild-type enzyme. Under microaerobic conditions, the ethanol production rates of the strain expressing the mutant xylose isomerase were considerably higher than previously reported values for yeast harboring a xylose isomerase pathway and were also comparable to those of the strains harboring an oxidoreductase pathway. Consequently, this study shows the potential to evolve a xylose isomerase pathway for more efficient xylose utilization.

scholarly journals Engineering of Saccharomyces cerevisiae for Efficient Anaerobic Alcoholic Fermentation of l-Arabinose

2007 ◽  
Vol 73 (15) ◽  
pp. 4881-4891 ◽  
Author(s):  
H. Wouter Wisselink ◽  
Maurice J. Toirkens ◽  
M. del Rosario Franco Berriel ◽  
Aaron A. Winkler ◽  
Johannes P. van Dijken ◽  
...  
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Ethanol Production ◽  
Alcoholic Fermentation ◽  
Plant Biomass ◽  
Ethanol Yield ◽  
Dry Weight ◽  
Pentose Phosphate ◽  
Anaerobic Growth ◽  
Evolutionary Engineering ◽  
First Case

ABSTRACT For cost-effective and efficient ethanol production from lignocellulosic fractions of plant biomass, the conversion of not only major constituents, such as glucose and xylose, but also less predominant sugars, such as l-arabinose, is required. Wild-type strains of Saccharomyces cerevisiae, the organism used in industrial ethanol production, cannot ferment xylose and arabinose. Although metabolic and evolutionary engineering has enabled the efficient alcoholic fermentation of xylose under anaerobic conditions, the conversion of l-arabinose into ethanol by engineered S. cerevisiae strains has previously been demonstrated only under oxygen-limited conditions. This study reports the first case of fast and efficient anaerobic alcoholic fermentation of l-arabinose by an engineered S. cerevisiae strain. This fermentation was achieved by combining the expression of the structural genes for the l-arabinose utilization pathway of Lactobacillus plantarum, the overexpression of the S. cerevisiae genes encoding the enzymes of the nonoxidative pentose phosphate pathway, and extensive evolutionary engineering. The resulting S. cerevisiae strain exhibited high rates of arabinose consumption (0.70 g h−1 g [dry weight]−1) and ethanol production (0.29 g h−1 g [dry weight]−1) and a high ethanol yield (0.43 g g−1) during anaerobic growth on l-arabinose as the sole carbon source. In addition, efficient ethanol production from sugar mixtures containing glucose and arabinose, which is crucial for application in industrial ethanol production, was achieved.

scholarly journals Novel Evolutionary Engineering Approach for Accelerated Utilization of Glucose, Xylose, and Arabinose Mixtures by Engineered Saccharomyces cerevisiae Strains

2008 ◽  
Vol 75 (4) ◽  
pp. 907-914 ◽  
Author(s):  
H. Wouter Wisselink ◽  
Maurice J. Toirkens ◽  
Qixiang Wu ◽  
Jack T. Pronk ◽  
Antonius J. A. van Maris
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Anaerobic Fermentation ◽  
Ethanol Yield ◽  
Cost Effective ◽  
Batch Cultivation ◽  
Evolutionary Engineering ◽  
Biotechnological Production ◽  
Production Processes ◽  
Sequential Fermentation ◽  
Time Required

ABSTRACT Lignocellulosic feedstocks are thought to have great economic and environmental significance for future biotechnological production processes. For cost-effective and efficient industrial processes, complete and fast conversion of all sugars derived from these feedstocks is required. Hence, simultaneous or fast sequential fermentation of sugars would greatly contribute to the efficiency of production processes. One of the main challenges emerging from the use of lignocellulosics for the production of ethanol by the yeast Saccharomyces cerevisiae is efficient fermentation of d-xylose and l-arabinose, as these sugars cannot be used by natural S. cerevisiae strains. In this study, we describe the first engineered S. cerevisiae strain (strain IMS0003) capable of fermenting mixtures of glucose, xylose, and arabinose with a high ethanol yield (0.43 g g−1 of total sugar) without formation of the side products xylitol and arabinitol. The kinetics of anaerobic fermentation of glucose-xylose-arabinose mixtures were greatly improved by using a novel evolutionary engineering strategy. This strategy included a regimen consisting of repeated batch cultivation with repeated cycles of consecutive growth in three media with different compositions (glucose, xylose, and arabinose; xylose and arabinose; and only arabinose) and allowed rapid selection of an evolved strain (IMS0010) exhibiting improved specific rates of consumption of xylose and arabinose. This evolution strategy resulted in a 40% reduction in the time required to completely ferment a mixture containing 30 g liter−1 glucose, 15 g liter−1 xylose, and 15 g liter−1 arabinose.

scholarly journals Effects of Ultrasound on Fermentation of Glucose to Ethanol by Saccharomyces cerevisiae

Fermentation ◽  
2019 ◽  
Vol 5 (1) ◽  
pp. 16 ◽  
Author(s):  
Luis Huezo ◽  
Ajay Shah ◽  
Frederick Michel
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Mass Transfer ◽  
Glucose Uptake ◽  
Ethanol Production ◽  
Ethanol Yield ◽  
Biofuel Production ◽  
Inhibitory Effects ◽  
Negative Effects ◽  
Intraparticle Mass Transfer ◽  
Improve Mass Transfer

Previous studies have shown that pretreatment of corn slurries using ultrasound improves starch release and ethanol yield during biofuel production. However, studies on its effects on the mass transfer of substrates and products during fermentation have shown that it can have both beneficial and inhibitory effects. In this study, the effects of ultrasound on mass transfer limitations during fermentation were examined. Calculation of the external and intraparticle observable moduli under a range of conditions indicate that no external or intraparticle mass transfer limitations should exist for the mass transfer of glucose, ethanol, or carbon dioxide. Fermentations of glucose to ethanol using Saccharomyces cerevisiae were conducted at different ultrasound intensities to examine its effects on glucose uptake, ethanol production, and yeast population and viability. Four treatments were compared: direct ultrasound at intensities of 23 and 32 W/L, indirect ultrasound (1.4 W/L), and no-ultrasound. Direct and indirect ultrasound had negative effects on yeast performance and viability, and reduced the rates of glucose uptake and ethanol production. These results indicate that ultrasound during fermentation, at the levels applied, is inhibitory and not expected to improve mass transfer limitations.

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

Fermentation ◽  
2018 ◽  
Vol 4 (3) ◽  
pp. 59 ◽  
Author(s):  
Tingting Liu ◽  
Shuangcheng Huang ◽  
Anli Geng
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Alcoholic Fermentation ◽  
Ethanol Yield ◽  
Xylose Isomerase ◽  
Cost Effective ◽  
Xylose Utilization ◽  
Yeast Strains ◽  
Multiple Copies ◽  
Biomass Sugars ◽  
Sugar Conversion

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.

scholarly journals Ethanol Production from Nondetoxified Dilute-Acid Lignocellulosic Hydrolysate by Cocultures of Saccharomyces cerevisiae Y5 and Pichia stipitis CBS6054

2012 ◽  
Vol 2012 ◽  
pp. 1-6 ◽  
Author(s):  
Ping Wan ◽  
Dongmei Zhai ◽  
Zhen Wang ◽  
Xiushan Yang ◽  
Shen Tian
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Ethanol Production ◽  
Ethanol Concentration ◽  
Ethanol Yield ◽  
Synthetic Medium ◽  
Pichia Stipitis ◽  
Dilute Acid ◽  
Acid Hydrolysate ◽  
Issatchenkia Orientalis ◽  
Final Ethanol Concentration

Saccharomyces cerevisiae Y5 (CGMCC no. 2660) and Issatchenkia orientalis Y4 (CGMCC no. 2159) were combined individually with Pichia stipitis CBS6054 to establish the cocultures of Y5 + CBS6054 and Y4 + CBS6054. The coculture Y5 + CBS6054 effectively metabolized furfural and HMF and converted xylose and glucose mixture to ethanol with ethanol concentration of 16.6 g/L and ethanol yield of 0.46 g ethanol/g sugar, corresponding to 91.2% of the maximal theoretical value in synthetic medium. Accordingly, the nondetoxified dilute-acid hydrolysate was used to produce ethanol by co-culture Y5 + CBS6054. The co-culture consumed glucose along with furfural and HMF completely in 12 h, and all xylose within 96 h, resulting in a final ethanol concentration of 27.4 g/L and ethanol yield of 0.43 g ethanol/g sugar, corresponding to 85.1% of the maximal theoretical value. The results indicated that the co-culture of Y5 + CBS6054 was a satisfying combination for ethanol production from non-detoxified dilute-acid lignocellulosic hydrolysates. This co-culture showed a promising prospect for industrial application.

scholarly journals ATPase-based implementation of enforced ATP wasting in Saccharomyces cerevisiae for improved ethanol production

2020 ◽  
Vol 13 (1) ◽  
Author(s):  
Ahmed Zahoor ◽  
Katrin Messerschmidt ◽  
Simon Boecker ◽  
Steffen Klamt
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Atpase Activity ◽  
Ethanol Production ◽  
Ethanol Yield ◽  
Volumetric Productivity ◽  
Anaerobic Growth ◽  
Control Strain ◽  
Genomic Integration ◽  
Expression Of Genes ◽  
Atp Generation

Abstract Background Enforced ATP wasting has been recognized as a promising metabolic engineering strategy to enhance the microbial production of metabolites that are coupled to ATP generation. It also appears to be a suitable approach to improve production of ethanol by Saccharomyces cerevisiae. In the present study, we constructed different S. cerevisiae strains with heterologous expression of genes of the ATP-hydrolyzing F1-part of the ATPase enzyme to induce enforced ATP wasting and quantify the resulting effect on biomass and ethanol formation. Results In contrast to genomic integration, we found that episomal expression of the αβγ subunits of the F1-ATPase genes of Escherichia coli in S. cerevisiae resulted in significantly increased ATPase activity, while neither genomic integration nor episomal expression of the β subunit from Trichoderma reesei could enhance ATPase activity. When grown in minimal medium under anaerobic growth-coupled conditions, the strains expressing E. coli’s F1-ATPase genes showed significantly improved ethanol yield (increase of 10% compared to the control strain). However, elevated product formation reduces biomass formation and, therefore, volumetric productivity. We demonstrate that this negative effect can be overcome under growth-decoupled (nitrogen-starved) operation with high and constant biomass concentration. Under these conditions, which mimic the second (production) phase of a two-stage fermentation process, the ATPase-expressing strains showed significant improvement in volumetric productivity (up to 111%) compared to the control strain. Conclusions Our study shows that expression of genes of the F1-portion of E. coli’s ATPase induces ATPase activity in S. cerevisiae and can be a promising way to improve ethanol production. This ATP-wasting strategy can be easily applied to other metabolites of interest, whose formation is coupled to ATP generation.

Ethanol Production from Different Substrates by a Flocculent Saccharomyces cerevisiae Strain

2009 ◽  
Vol 7 (1) ◽  
Author(s):  
José Duarte ◽  
Vera Lourenço ◽  
Belina Ribeiro ◽  
Maria Céu Saagua ◽  
Joana Pereira ◽  
...  
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Ethanol Production ◽  
Yeast Strain ◽  
Fossil Fuels ◽  
Ethanol Yield ◽  
Corn Fiber ◽  
Production Costs ◽  
Raw Material ◽  
Continuous Reactor ◽  
Different Strains

During the last years there has been an increasing interest in using ethanol as a substitute for fossil fuels. The bioethanol used today is mainly produced from sugar cane and cereals, but reducing the production costs of ethanol is still crucial for a viable economic process. Cellulose from vegetable biomass will be the next cheap raw material for second generation fuel ethanol production and agricultural by-products with a low commercial value, as corn stover, corn fiber and cane bagasses would become an attractive feedstock for bioethanol production.In this study, different strains of Saccharomyces cerevisiae have been screened for the ability of bioethanol production. Yeasts were grown in a synthetic liquid medium containing sucrose in batch regime and the growth rates, ethanol and biomass productions were determined as well as their growth ability in cane molasses.The results indicate that a flocculent yeast, isolated in our lab and designated by strain F, was the most promising yeast strain among those tested for continuous ethanol production. This strain was isolated from corn hydrolysates, obtained from a Portuguese distillery facility (DVT, Torres Novas, Portugal) showing highest growth rate (0.49h-1), highest ethanol yield (0.35g/g) and high flocculation capacity.The study on ethanol production in continuous reactor process with the selected yeast strain (strain F) was made on sucrose and cane molasses at different dilution rates (0.05-0.42 h-1). A steady flocculating yeast fluidized bed reactor system was established allowing the functioning of the reactor for 1000 h. Data shows that when the dilution rate rose to 0.42h-1, the highest productivity (20g/Lh) was obtained attaining an ethanol concentration in the reactor of 47g/L for sucrose and molasses media.

scholarly journals Investigation of the toxicity of the ionic liquid 1-butyl-3-methylimidazolium chloride to Saccharomyces cerevisiae AY93161 for lignocellulosic ethanol production

2013 ◽  
Vol 15 (2) ◽  
pp. 94-98 ◽  
Author(s):  
Shengdong Zhu ◽  
Pei Yu ◽  
Mingke Lei ◽  
Yanjie Tong ◽  
Lu Zheng ◽  
...  
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Ionic Liquid ◽  
Ethanol Production ◽  
Metabolic Activity ◽  
Morphological Structure ◽  
Optical Microscope ◽  
Reproduction Rate ◽  
Lignocellulosic Ethanol ◽  
Lignocellulosic Materials ◽  
Liquid Suspension

Ionic liquid (IL) pretreatment of lignocellulosic materials has provided a new technical tool to improve lignocellulosic ethanol production. To evaluate the influence of the residual IL in the fermentable sugars from enzymatic hydrolysis of IL pretreatment of lignocellulosic materials on the subsequent ethanol fermentation, the toxicity of the IL 1-butyl-3-methylimidazolium chloride ([BMIM]Cl) to Saccharomyces cerevisiae AY93161 was investigated. Firstly, the morphological structure, budding and metabolic activity of Saccharomyces cerevisiae AY93161 at different [BMIM]Cl concentrations were observed under an optical microscope. The results show that its single cell morphology remained unchanged at all [BMIM]Cl concentrations, but its reproduction rate by budding and its metabolic activity decreased with the [BMIM]Cl concentration increasing. The half effective concentration (EC50) and the half inhibition concentration (IC50) of [BMIM]Cl to Saccharomyces cerevisiae AY93161 were then measured using solid and liquid suspension culture and their value were 0.53 and 0.39 g.L-1 respectively. Finally, the influence of [BMIM]Cl on ethanol production was investigated. The results indicate that the [BMIM]Cl inhibited the growth and ethanol production of Saccharomyces cerevisiae AY93161. This toxicity study provides useful basic data for further development in lignocellulosic ethanol production by using IL technology and it also enriches the IL toxicity data.

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