Chemical mutagenesis of Saccharomyces cerevisiae for enhancing bioethanol production with fermentation at very high sugar concentration

BioResources ◽  
2020 ◽  
Vol 15 (1) ◽  
pp. 1354-1369
Author(s):  
Nadeem I. Elhussieny ◽  
Marwah M. Bakri ◽  
Magdah Ganash ◽  
Tarek M. Abdel Ghany
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Ethanol Production ◽  
Incubation Period ◽  
Dna Analysis ◽  
Ethanol Yield ◽  
Random Amplified Polymorphic Dna ◽  
Chemical Mutagenesis ◽  
Bioethanol Production ◽  
High Sugar ◽  
Very High

Saccharomyces cerevisiae is one of the most promising unicellular fungi on account of its vital applications in biotechnology as well as bioethanol production. Improvement of ethanol production via very high-gravity (VHG) fermentation (fermentation at high sugar levels) was successfully developed using the ethidium bromide (EtB) mutagenesis of S. cerevisiae. This study found two developed mutants of S. cerevisiae (EtB20a and EtB20b) with varied capacity for ethanol production using EtB, depending on random amplified polymorphic DNA analysis. Mutant EtB20b showed improved ethanol yield (19.5%) compared with the wild-type (18.0%), while the other mutant EtB20a exhibited retarded ethanol production (9.1%). Optimization of ethanol production by mutant EtB20b was performed under other conditions including temperature, pH, inoculum size, and incubation period. The highest production capacity of the yeasts was 20.8, 19.9, 19.5, and 19.5% at an optimum temperature of 30 °C, pH 6.0, incubation period of 72 h, and 1 mL of yeast suspension (optical density at 600 nm) with glucose utilization of 42.6, 40.7, 39.8, and 39.9%, respectively.

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 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.

Bioethanol Production from Pineapple Peel Juice Using Saccharomyces cerevisiae

2014 ◽  
Vol 875-877 ◽  
pp. 242-245
Author(s):  
Jutarut Pornpunyapat ◽  
Wilaiwan Chotigeat ◽  
Pakamas Chetpattananondh
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Ethanol Production ◽  
Ethanol Concentration ◽  
Renewable Resource ◽  
Reducing Sugar ◽  
Total Sugar ◽  
Bioethanol Production ◽  
Pineapple Peel ◽  
Initial Ph ◽  
Production Characteristics

Bioethanol is widely used as renewable resource due to its safe to produce and environmentally friendly. However, knowledge on ethanol production from pineapple peel juice (Pattawia spp) is far from sufficient. In this work, pineapple peel juice (initial pH at 5) was fermented at various yeast contents (1, 3 and 5% by wt.) and fermentation times (3, 5 and 7 days) in order to investigate ethanol production characteristics. Yeast, Sacchromyces cerevisiae was grown on pineapple peel juice. The squeezed juice contained 11% of total sugar and 5% of reducing sugar. The results indicated that the optimum ethanol production was yeast contents of 5% by wt. and fermentation times of 5 days which gave the ethanol production of 9.08g/l. The ethanol at a higher yeast content also had a higher ethanol concentration.

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 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 Efficient Bioethanol Production by a Recombinant Flocculent Saccharomyces cerevisiae Strain with a Genome-Integrated NADP+-Dependent Xylitol Dehydrogenase Gene

2009 ◽  
Vol 75 (11) ◽  
pp. 3818-3822 ◽  
Author(s):  
Akinori Matsushika ◽  
Hiroyuki Inoue ◽  
Seiya Watanabe ◽  
Tsutomu Kodaki ◽  
Keisuke Makino ◽  
...  
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Ethanol Yield ◽  
Xylitol Dehydrogenase ◽  
Wood Chips ◽  
Bioethanol Production ◽  
Xylose Consumption ◽  
Acid Hydrolysate ◽  
A Genome ◽  
Dehydrogenase Gene ◽  
Flocculent Yeast

ABSTRACT The recombinant industrial Saccharomyces cerevisiae strain MA-R5 was engineered to express NADP+-dependent xylitol dehydrogenase using the flocculent yeast strain IR-2, which has high xylulose-fermenting ability, and both xylose consumption and ethanol production remarkably increased. Furthermore, the MA-R5 strain produced the highest ethanol yield (0.48 g/g) from nonsulfuric acid hydrolysate of wood chips.

Use of sugarcane molasses “B” as an alternative for ethanol production with wild-type yeast Saccharomyces cerevisiae ITV-01 at high sugar concentrations

2011 ◽  
Vol 35 (4) ◽  
pp. 605-614 ◽  
Author(s):  
C. L. Fernández-López ◽  
B. Torrestiana-Sánchez ◽  
M. A. Salgado-Cervantes ◽  
P. G. Mendoza García ◽  
M. G. Aguilar-Uscanga
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Ethanol Production ◽  
Sugarcane Molasses ◽  
Wild Type ◽  
Yeast Saccharomyces Cerevisiae ◽  
High Sugar ◽  
Wild Type Yeast

scholarly journals Evaluating and engineering Saccharomyces cerevisiae promoters for increased amylase expression and bioethanol production from raw starch

2020 ◽  
Vol 20 (6) ◽  
Author(s):  
Marthinus W Myburgh ◽  
Shaunita H Rose ◽  
Marinda Viljoen-Bloom
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Corn Starch ◽  
Consolidated Bioprocessing ◽  
Ethanol Yield ◽  
Small Scale ◽  
Bioethanol Production ◽  
Raw Starch ◽  
Broken Rice ◽  
Available Carbon ◽  
Theoretical Ethanol Yield

ABSTRACT Bioethanol production from starchy biomass via consolidated bioprocessing (CBP) will benefit from amylolytic Saccharomyces cerevisiae strains that produce high levels of recombinant amylases. This could be achieved by using strong promoters and modification thereof to improve gene expression under industrial conditions. This study evaluated eight endogenous S. cerevisiae promoters for the expression of a starch-hydrolysing α-amylase gene. A total of six of the native promoters were modified to contain a promoter-proximal intron directly downstream of the full-length promoter. Varying results were obtained; four native promoters outperformed the ENO1P benchmark under aerobic conditions and two promoters showed better expression under simulated CBP conditions. The addition of the RPS25A intron significantly improved the expression from most promoters, displaying increased transcript levels, protein concentrations and amylase activities. Raw starch-utilising strains were constructed through co-expression of selected α-amylase cassettes and a glucoamylase gene. The amylolytic strains displayed improved fermentation vigour on raw corn starch and broken rice, reaching 97% of the theoretical ethanol yield and converting 100% of the available carbon to products within 120 h in small-scale CBP fermentations on broken rice. This study showed that enhanced amylolytic strains for the conversion of raw starch to ethanol can be achieved through turnkey promoter selection and/or engineering.

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