Search

A previously published genome‐scale metabolic model namely iFF708 was modified to depict the metabolic flux distribution within Saccharomyces cerevisiae grown under a redox potential–controlled very high‐gravity condition. The following modifications were made: electron transport chain (ETC) and oxidative phosphorylation, proton gradient and ATP transportation, and malate–aspartate shuttle. With these modifications, this model could describe the experimental data collected from the above‐mentioned ethanol fermentation. As a result, the simulation unveiled that the P/O ratio is critical under microaerobic conditions and the malate–aspartate shuttle is inactivated due to the shortage of electron transport across mitochondria. In other words, the limited supply of oxygen suppresses the functionality of oxidative phosphorylation, tricarboxylic acid (TCA) cycle, and ETC. In terms of the glycolytic pathway, fluxes coming from glucose‐6‐phosphate and pyruvate nodes are insensitive to the changes of fermentation redox potential. As the initial glucose concentration is greater than 250 g/L, the interactive effect between the initial glucose concentration and redox potential level becomes noticeable. [ABSTRACT FROM AUTHOR]

Trehalase catalyzes the hydrolysis of the non-reducing disaccharide trehalose. The highly active trehalase MthT from Myceliophthora thermophila was screened from the trehalase genes of six species of filamentous fungi. An ingenious multi-copy knock-in expression strategy mediated by the CRISPR/Cas9 tool and medium optimization were used to improve MthT production in Aspergillus niger, up to 1698.83 U/mL. The protein background was dramatically abated due to insertion. The recombinant MthT showed optimal activity at pH 5.5 and 60 °C, and exhibited prominent thermal stability between 50 and 60 °C under acid conditions (pH 4.5–6.5). The ethanol conversion rate (ethanol yield/total glucose) was significantly improved by addition of MthT (51.88%) compared with MthT absence (34.38%), using 30% starch saccharification liquid. The results of this study provided an effective strategy, established a convenient platform for heterologous expression in A. niger and showed a potential strategy to decrease production costs in industrial ethanol production. [ABSTRACT FROM AUTHOR]

Abstract Background High temperature inhibits cell growth and ethanol fermentation of Saccharomyces cerevisiae. As a complex phenotype, thermotolerance usually involves synergistic actions of many genes, thereby being difficult to engineer. The overexpression of either endogenous or exogenous stress-related transcription factor genes in yeasts was found to be able to improve relevant stress tolerance of the hosts. Results To increase ethanol yield of high-temperature fermentation, we constructed a series of strains of S. cerevisiae by expressing 8 transcription factor genes from S. cerevisiae and 7 transcription factor genes from thermotolerant K. marxianus in S. cerevisiae. The results of growth curve measurements and spotting test show that KmHsf1 and KmMsn2 can enhance cell growth of S. cerevisiae at 40–42 °C. According to the results of batch fermentation at 43 °C with an initial glucose concentration of 104.8 g/l, the fermentation broths of KmHSF1 and KmMSN2-expressing strains could reach final ethanol concentrations of 27.2 ± 1.4 and 27.6 ± 1.2 g/l, respectively, while the control strain just produced 18.9 ± 0.3 g/l ethanol. Transcriptomic analysis found that the expression of KmHSF1 and KmMSN2 resulted in 55 (including 31 up-regulated and 24 down-regulated) and 50 (including 32 up-regulated and 18 down-regulated) genes with different expression levels, respectively (padj

Abstract Background Multiple lignocellulose-derived inhibitors represent great challenges for bioethanol production from lignocellulosic materials. These inhibitors that are related to the levels of intracellular reactive oxidative species (ROS) make oxidoreductases a potential target for an enhanced tolerance in yeasts. Results In this study, the thioredoxin and its reductase from Kluyveromyces marxianus Y179 was identified, which was subsequently achieved over-expression in Saccharomyces cerevisiae 280. In spite of the negative effects by expression of thioredoxin gene (KmTRX), the thioredoxin reductase (KmTrxR) helped to enhance tolerance to multiple lignocellulose-derived inhibitors, such as formic acid and acetic acid. In particular, compared with each gene expression, the double over-expression of KmTRX2 and KmTrxR achieved a better ethanol fermentative profiles under a mixture of formic acid, acetic acid, and furfural (FAF) with a shorter lag period. At last, the mechanism that improves the tolerance depended on a normal level of intracellular ROS for cell survival under stress. Conclusions The synergistic effect of KmTrxR and KmTRX2 provided the potential possibility for ethanol production from lignocellulosic materials, and give a general insight into the possible toxicity mechanisms for further theoretical research.

In this study chemical and enzymatic treatment of hemp biomass were optimized to obtain maximum ethanol production. In the first stage, physical and chemical pretreatment of hemp biomass was carried out. It was found that the Tygra variety is susceptible to alkaline treatment at an optimum concentration of 2% NaOH. Next, the effect of NaOH on the value of reducing sugars and the chemical composition of the solid fraction before and after the treatment was determined. Hemp biomass before and after the chemical treatment was analysed by FTIR spectra and SEM. The effect of enzymatic hydrolysis, i.e., substrate content, temperature, time, pH and dose of enzyme by means of Response Surface Methodology on glucose content was determined. The highest glucose value was observed at 50 °C, in time process between 48 and 72 h, and the dose of enzyme was not less than 20 FPU·g−1. After the optimization of enzymatic hydrolysis two processes of ethanol fermentation from hemp biomass, SHF and SSF, were carried out. In the SHF process a 40% higher concentration of ethanol was obtained (10.51 g/L). In conclusion, hemp biomass was found to be an interesting and promising source to be used for bioethanol production. [ABSTRACT FROM AUTHOR]

Biofuel research and development roadmap is currently underway in several countries and is expected to pave a way for the establishment of a viable renewable energy sector that can compete with petroleum-based fuels. Ethanol fermentation has garnered increasing attention amongst various stakeholders (industries, governments, and academia) due to its economic and environmental merits. However, microbial contamination continues to be one of the major barriers in ethanologenic processes, resulting in low ethanol yields and thereby translating into economic losses. To this end, technological innovations geared towards effective elimination of microbial contamination are constantly being developed. This review explores and discusses the fermentation conditions that facilitate the growth of undesired microorganisms during ethanol fermentation processes. It highlights the methods that are currently used in biorefineries as well as innovative and advanced biotechnological methods currently being evaluated as viable alternative strategies to control or eliminate microbial contaminants in ethanol fermentations. These methods have the potential to minimize or control the contamination problem and could pave a way for the development of an efficient biofuel sector. [ABSTRACT FROM AUTHOR]

Herbicides inhibiting either aromatic or branched-chain amino acid biosynthesis trigger similar physiological responses in plants, despite their different mechanism of action. Both types of herbicides are known to activate ethanol fermentation by inducing the expression of fermentative genes; however, the mechanism of such transcriptional regulation has not been investigated so far. In plants exposed to low-oxygen conditions, ethanol fermentation is transcriptionally controlled by the ethylene response factors-VII (ERF-VIIs), whose stability is controlled in an oxygen-dependent manner by the Cys-Arg branch of the N-degron pathway. In this study, we investigated the role of ERF-VIIs in the regulation of the ethanol fermentation pathway in herbicide-treated Arabidopsis plants grown under aerobic conditions. Our results demonstrate that these transcriptional regulators are stabilized in response to herbicide treatment and are required for ethanol fermentation in these conditions. We also observed that mutants with reduced fermentative potential exhibit higher sensitivity to herbicide treatments, thus revealing the existence of a mechanism that mimics oxygen deprivation to activate metabolic pathways that enhance herbicide tolerance. We speculate that this signaling pathway may represent a potential target in agriculture to affect tolerance to herbicides that inhibit amino acid biosynthesis. [ABSTRACT FROM AUTHOR]

• Integration of a green biorefinery with ethanol fermentation was evaluated. • 90% of the green juice crude protein was recovered in the protein concentrate. • Nanofiltration raised the sugar composition in the brown juice from 2.4 to 13.6 %. • Purification of ethanol from concentrated brown juice used up to 45% less energy. • DWC was the best choice to purify ethanol among the tested extractive distillations. The objective of this study is to propose the integration of a green protein biorefinery with ethanol production and to highlight the potential of nanofiltration membrane technologies and extractive distillation in the separation and purification of valuable substances. Depending on the product and its purity, the nanofiltration membrane can help in decreasing energy expenditure and greenhouse gas emissions while extractive distillation overcomes the water–ethanol azeotrope formation. The aim is to enhance the sustainability of biorefinery processes by integrating new separation techniques and adopting process intensification. The concentration of the residual brown juice using nanofiltration reduced its water content and favored the sugar conversion into ethanol while concomitantly decreasing the formation of byproducts. It has been shown that these configurations (with the presence of nanofiltration and intensified) have lower energy consumption compared to the proposed classical sequence. The results of the simulations indicate that better results are obtained with the extractive distillation using a dividing-wall column, achieving around a 50 % reduction in energy consumption and a 58 % reduction in CO 2 emissions. [ABSTRACT FROM AUTHOR]

Bio-ethanol 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 plant biomass will be the next cheaper raw material for second generation fuel ethanol production. Saccharomyces cerevisiae and Schizosaccharomyces pombae are the common industrial yeast strains used for the production of ethanol from various substrates derived from agricultural commodities. In the present study, ethanol was produced from various substrates such as molasses, sugarcane juice, sugarcane syrup, jaggery, grapes, beet root, orange and rice substrates by using Saccharomyces cerevisiae and Schizosaccharomyces pombae. The experimental study shows that the biomass production declines progressively when the ethanol accumulates in the surrounding medium after the production of ethanol. However, the conversion of ethanol was achieved in batch fermentation process at optimum temperature and the internal pH of the fermentation medium at laboratory level. The yield percentage of ethanol for the above substrates were analyzed and compared for different carbon sources. Among different substrates, Jaggery and Molases showed higher ethanol yield as 95.2% and 93.5% respectively. Among the two species of yeast studied, Saccharomyces cerevisiae yielded higher percentage of ethanol than Schizosaccharomyces pombae in batch fermentation process. The present study concludes the effect of various carbon sources on production of ethanol with two different industrial yeast strains such as Saccharomyces cerevisiae and Schizosaccharomyces pombae are vary in similar conditions of ethanol production processprocess.

Objectives: To improve ethanolic fermentation performance of self-flocculating yeast, difference between a flocculating yeast strain and a regular industrial yeast strain was analyzed by transcriptional and metabolic approaches. Results: The number of down-regulated (industrial yeast YIC10 vs. flocculating yeast GIM2.71) and up-regulated genes were 4503 and 228, respectively. It is the economic regulation for YIC10 that non-essential genes were down-regulated, and cells put more “energy” into growth and ethanol production. Hexose transport and phosphorylation were not the limiting-steps in ethanol fermentation for GIM2.71 compared to YIC10, whereas the reaction of 1,3-disphosphoglycerate to 3-phosphoglycerate, the decarboxylation of pyruvate to acetaldehyde and its subsequent reduction to ethanol were the most limiting steps. GIM2.71 had stronger stress response than non-flocculating yeast and much more carbohydrate was distributed to other bypass, such as glycerol, acetate and trehalose synthesis. Conclusions: Differences between flocculating yeast and regular industrial yeast in transcription and metabolite profiling will provide clues for improving the fermentation performance of GIM2.71.

Abstract Background Bioethanol from lignocellulosic materials is of great significance to the production of renewable fuels due to its wide sources. However, multiple inhibitors generated from pretreatments represent great challenges for its industrial-scale fermentation. Despite the complex toxicity mechanisms, lignocellulose-derived inhibitors have been reported to be related to the levels of intracellular reactive oxygen species (ROS), which makes oxidoreductase a potential target for the enhancement of the tolerance of yeasts to these inhibitors. Results A typical 2-Cys peroxiredoxin from Kluyveromyces marxianus Y179 (KmTPX1) was identified, and its overexpression was achieved in Saccharomyces cerevisiae 280. Strain TPX1 with overexpressed KmTPX1 gene showed an enhanced tolerance to oxidative stresses. Serial dilution assay indicated that KmTPX1 gene contributed to a better cellular growth behavior, when the cells were exposed to multiple lignocellulose-derived inhibitors, such as formic acid, acetic acid, furfural, ethanol, and salt. In particular, KmTPX1 expression also possessed enhanced tolerance to a mixture of formic acid, acetic acid, and furfural (FAF) with a shorter lag period. The maximum glucose consumption rate and ethanol generation rate in KmTPX1-expressing strain were significantly improved, compared with the control. The mechanism of improved tolerance to FAF depends on the lower level of intracellular ROS for cell survival under stress. Conclusion A new functional gene KmTPX1 from K. marxianus is firstly associated with the enhanced tolerance to multiple lignocellulose-derived inhibitors in S. cerevisiae. We provided a possible detoxification mechanism of the KmTPX1 for further theoretical research; meanwhile, we provided a powerful potential for application of the KmTPX1 overexpressing strain in ethanol production from lignocellulosic materials.

Methane and ethanol were co-produced from different feedstock, including a mixture of dairy manure and soybean straw (DMS), a mixture of pig manure and soybean straw (PMS), and soybean straw alone (SS), after anaerobic digestion times of 30 and 60 days in mesophilic conditions. Digesting DMS for 60 days led to the highest methane yield of 115.3 g/kg dry raw feed; however, the lowest ethanol yield of 88 g/kg dry raw feed was observed. After 30 days, SS yielded the lowest methane levels (45.2 g/kg dry raw feed) but the highest ethanol levels (113.5 g/kg dry raw feed). Analysis of the net energy balance showed that the highest net energy balance, 6549 kJ/kg of dry raw feedstock, was achieved from the digestion of DMS for 60 days. Overall, both the type of feedstock and length of digestion time played important roles in the integrated processing of methane and ethanol from livestock manure and straw.

This paper investigates the control of an experimentally validated model of production of bioethanol. The analysis of the open loop system revealed that the maximum productivity occurred at a periodic point. A robust control was needed to avoid instabilities that may occur when disturbances are injected into the process that may drive it toward or through the unstable points. A nonlinear model predictive controller (NLMPC) was used to control the process. Simulation tests were carried out using three controlled variables: the ethanol concentration, the productivity and the inverse of the productivity. In the third configuration, the controller was required to seek the maximum operating point through the optimization capability built in the NLMPC algorithm. Simulation tests presented overall satisfactory closed-loop performance for both nominal servo and regulatory control problems as well as in the presence of modeling errors. The third control configuration managed to steer the process toward the existing maximum productivity even when the process operation or its parameters changed. For comparison purposes, a standard PI controller was also designed for the same control objectives. The PI controller yielded satisfactory performance when the ethanol concentration was chosen as the controlled variable. When, on the other hand, the productivity was chosen as the controlled output, the PI controller did not work properly and needed to be adjusted using gain scheduling. In all cases, it was observed that the closed-loop response suffered from slow dynamics, and any attempt to speed up the feedback response via tuning may result in an unstable behavior.

To make biofuel production feasible from an economic point of view, several studies have investigated the main associated bottlenecks of the whole production process through approaches such as the “cradle to grave” approach or the Life Cycle Assessment (LCA) analysis, being the main constrains the feedstock collection and transport. Whilst several feedstocks are interesting because of their high sugar content, very few of them are available all year around and moreover do not require high transportation’ costs. This work aims to investigate if the “zero miles” concept could bring advantages to biofuel production by decreasing all the associated transport costs on a locally established production platform. In particular, a specific case study applied to the Technical University of Denmark (DTU) campus is used as example to investigate the advantages and feasibility of using the spent coffee grounds generated at the main cafeteria for the production of bioethanol on site, which can be subsequently used to (partially) cover the campus’ energy demands.

This study reports on the evaluation of a two-stage pretreatment process for preparing cotton gin trash (CGT) for conversation to ethanol. During the first stage, CGT was mixed with acid at 12% H2SO4 on solids and heated to 180 °C for 15 min in a pressurised stirred reactor. Pressed first-stage pretreated fibres were heated to 200 °C for 5 min during the second stage. The two-stage process facilitated excellent sugar recovery from both cellulose (84%) and hemicellulose (78%) fractions of CGT. Recombinant yeast GSF335 propagated on first-stage liquors yielded 41.8 g kg−1 of dry CGT and was compared with the commercial yeast during separate hydrolysis and fermentation (SHF) and simultaneous saccharification and fermentations (SSF). SHF of pretreated CGT fibre yielded the equivalent of 66.2 and 64.0 kg ethanol per tonne of unprocessed CGT with GSF335 and the commercial yeast, respectively. SSF ethanol yields for the commercial yeast were significantly lower (50.3 kg) while GSF335 correspondingly produced 63.8 kg ethanol per tonne of raw CGT. [ABSTRACT FROM AUTHOR]

• Humic substances (HS) consisted mainly of polyphenols and polysaccharides. • HS derived from rice straw promoted tomato growth. • HS exhibited an inhibitory effect on saccharification and fermentation. • HS production integrated with ethanol fermentation is a strategy for straw utilization. The aim of this study was to investigate the effects of bioactives derived from rice straw on tomato growth and develop an integrated process for utilization of straw. The humic substances (HS) was obtained from rice straw in this study. To investigate the effect of HS on tomato growth, tomato seedlings were sprayed with HS and the changes in morphology were measured. Compared with the control, the relative stem heights and leaf numbers of the tomato plants increased by 17.05% and 23.47% for polyphenols in HS and by 28.27% and 17.66% for polysaccharides in HS, respectively. Moreover, the inhibition of HS on cellulase activity and ethanol fermentation were confirmed in this study. When HS was added, the activities of FPase, CMCase and β-glucosidase decreased by 37.29%, 11.54% and 30.70%, respectively. Essentially, the production of HS could be considered as a removal step prior to the bioconversion of the straw into ethanol, and it increased the productivity of ethanol fermentation by 29.39%. The findings demonstrated that HS from rice straw could be successfully used as an environmentally friendly product to increase plant yield and, the residue after HS removal exhibited an improved efficiency of bioconversion to ethanol, making biofuels more attractive. [ABSTRACT FROM AUTHOR]

SPs could alleviate inhibition of phenolic acids, leading to high fermentability. • Soluble polysaccharides (SPs) were obtained from rice straw. • Xylan and glucan were the main components of SPs. • SPs enhanced the formation of polysaccharide-dependent extracellular matrix. • Extracellular matrix secreted by Saccharomyces cerevisiae maintained cellular integrity even under phenolic acid stress. • SPs increased ethanol production by S. cerevisiae. The bioconversion of rice straw into ethanol can alleviate the energy crisis and solve problems related to waste treatment. In this study, the effect of soluble polysaccharides (SPs) produced during rice straw saccharification on the formation of extracellular matrices (EMs) by the yeast Saccharomyces cerevisiae was investigated. SPs were characterized by high-performance liquid chromatography (HPLC) and fourier transform infrared spectroscopy (FT-IR). SPs reduced the inhibition of alcohol dehydrogenase activity by phenolic acids (PAs) and regulated the intracellular redox state, resulting in higher ethanol production. The results of flow cytometry, confocal laser scanning microscopy, and atomic force microscopy indicated that PAs changed microbial morphology and caused damage in microbial cell membranes. The protective effect of SPs against cell membrane damage could be attributed to the synthesis of polysaccharide-dependent extracellular matrix, which maintained cellular integrity even under phenolic acid stress. These findings provide new strategies to improve pretreatment and saccharification processes. [ABSTRACT FROM AUTHOR]

Wheat sourdough is a common traditional fermented food that is produced worldwide. However, product quality of spontaneous sourdough is not easy to control because it depends on natural fermentation and backslopping, about which little is known, notably after ten backslopping steps. To this end, we tracked the spontaneous fermentation of three sourdoughs made from wheat flours during 32 backslopping steps for 60 days. At 24 time points, the microbial community was analyzed by both culture-dependent and culture-independent methods and its chemical constituents were assessed. Dynamic changes were observed in the microbial community, which showed a common succession pattern among the three sourdoughs at the bacterial family level and differences at the species level. The bacterial communities evolved through three phases that were driven by different groups of lactic acid bacteria (LAB) species. The dynamism among the metabolites also differed, depending on the species composition of the LAB and yeast communities. In one sourdough, the growth of Saccharomyces cerevisiae was detected along with a concentration of increased ethanol, while in the other two sourdoughs, Wickerhamomyces anomalus was detected without ethanol production. Regarding the LAB communities, two sourdoughs were eventually co-dominated by Lactobacillus plantarum and L. brevis , while the other sourdough was eventually dominated solely by the heterolactic fermentative bacterium Lactobacillus fermentum , and ethanol was produced at the same level as lactic acid. Further research is needed to determine the bacterial and yeast species involved in the fermentation of sourdough, to help improve the design and quality control of the final product. • The first report to track spontaneous sourdough in detail for 60 days. • Lactobacillus fermentum stabilized heterolactic fermentation at late stage. • Lactic acid increases until the final when two Lactobacillus species co-dominate. • Ethanol increases with sugars exhaustion when Saccharomyces cerevisiae is retrieved. [ABSTRACT FROM AUTHOR]

N-Acetyltransferase Mpr1 was originally discovered as an enzyme that detoxifies l-azetidine-2-carboxylate through its N-acetylation in the yeast Saccharomyces cerevisiae Σ1278b. Mpr1 protects yeast cells from oxidative stresses possibly by activating a novel l-arginine biosynthesis. We recently constructed a stable variant of Mpr1 (N203K) by a rational design based on the structure of the wild-type Mpr1 (WT). Here, we examined the effects of N203K on ethanol fermentation of the sake yeast S. cerevisiae strain lacking the MPR1 gene. When N203K was expressed in the diploid Japanese sake strain, its fermentation performance was improved compared to WT. In a laboratory-scale brewing, a sake strain expressing N203K produced more ethanol than WT. N203K also affected the contents of flavor compounds and organic acids. These results suggest that the stable Mpr1 variant contributes to the construction of new industrial yeast strains with improved fermentation ability and diversity of taste and flavor. [ABSTRACT FROM AUTHOR]

A thermotolerant ethanol fermenting yeast strain is a key requirement for effective ethanol production at high temperature. This work aimed to select a thermotolerant yeast producing a high ethanol concentration from molasses and increasing its ethanol production by mutagenesis. Saccharomyces cerevisiae DMKU 3-S087 was selected from 168 ethanol producing strains because it produced the highest ethanol concentration from molasses at 40 °C. Optimization of molasses broth composition was performed by the response surface method using Box–Behnken design. In molasses broth containing optimal total fermentable sugars (TFS) of 200 g/L and optimal (NH4)2SO4 of 1 g/L, with an initial pH of 5.5 by shaking flask cultivation at 40 °C ethanol, productivity and yield were 58.4 ± 0.24 g/L, 1.39 g/L/h and 0.29 g/g, respectively. Batch fermentation in a 5 L stirred-tank fermenter with 3 L optimized molasses broth adjusted to an initial pH of 5.5 and fermentation controlled at 40 °C and 300 rpm agitation resulted in 72.4 g/L ethanol, 1.21 g/L/h productivity and 0.36 g/g yield at 60 h. Strain DMKU 3-S087 improvement was performed by mutagenesis using ultraviolet radiation and ethyl methane sulfonate (EMS). Six EMS mutants produced higher ethanol (65.2 ± 0.48–73.0 ± 0.54 g/L) in molasses broth containing 200 g/L TFS and 1 g/L (NH4)2SO4 by shake flask fermentation at 37 °C than the wild type (59.8 ± 0.25 g/L). Among these mutants, only mutant S087E100-265 produced higher ethanol (62.5 ± 0.26 g/L) than the wild type (59.5 ± 0.02 g/L) at 40 °C. In addition, mutant S087E100-265 showed better tolerance to high sugar concentration, furfural, hydroxymethylfurfural and acetic acid than the wild type. [ABSTRACT FROM AUTHOR]

Abstract Very high-gravity (VHG) worts (24 °P) supplemented with wheat gluten hydrolysates (WGH) and their ultrafiltration fractions (Mw > 1 kDa and <1 kDa fractions) were used to evaluate their effects on physiological activity and ethanol fermentation of brewer's yeast. The results demonstrated that WGH and their ultrafiltration fractions supplementations could obviously improve the biomass and viability of yeast, and further enhance the sugar utilization and ethanol production under VHG fermentation. Supplementation with Mw < 1 kDa fraction exhibited the maximum biomass (6.90 g/L dry cell), sugar utilization (79.79%) and ethanol concentration (11.88%, v/v) during VHG fermentation. Meanwhile, the addition of Mw < 1 kDa fraction significantly decreased the plasma membrane permeability and intracellular reactive oxygen species (ROS) accumulation, and increased the mitochondrial membrane potential in cells during VHG fermentation. Moreover, Mw < 1 kDa fraction supplementation caused the highest accumulations of intracellular trehalose and glycerol, which were 12.10% and 31.22% higher than the control, respectively. These results illustrated that WGH (especially Mw < 1 kDa fraction) addition was an economic and efficient method to improve physiological activity and ethanol fermentation of brewer's yeast in VHG worts. Highlights • WGH and their ultrafiltration fractions were obtained as nitrogen sources of yeast. • WGH supplementation changed nitrogen metabolism of yeast during VHG fermentation. • Biomass and ethanol production were increased by WGH addition under VHG condition. • Physiological activity of yeast was improved by WGH addition under VHG condition. • Trehalose and glycerol in yeast accumulated by WGH addition under VHG condition. [ABSTRACT FROM AUTHOR]

Abstract In this study, edible filamentous fungi Aspergillus oryzae and Rhizopus oligosporus were co-cultured with fodder yeast Candida utilis using corn thin stillage to produce feed-grade biomass, amylases, and ethanol with the purpose of backset valorization. The co-culture of A. oryzae with C. utilis inoculated after 24 h of pure fungal growth provided the most promising results, yielding 18.6 g L−1 biomass with 41 and 31% of lipid and protein, respectively, with all of the essential amino acids. The microorganisms consumed nearly 34% of the cultivation medium solids, as well as 33% of glycerol and 12% of lactic acid, which allowed them to produce ca. 4.5 g L−1 ethanol and 1.7 U mL−1 amylase. The liquid fraction of the cultivation medium was used as 200 mL L−1 backset for the production of high-solids corn mash. The final ethanol concentrations reached 78.3, 90.6 and 108.2 g L−1 in the control sample (without backset), unfermented backset addition and fermented backset addition, respectively. This suggests that the yeast-fungi co-culture for backset treatment was effective in improving the core production stage in an ethanol-producing plant. Highlights • Co-culture of fungi and yeast on thin stillage for backset valorization was studied. • 33.9% solids, 12.5% lactic acid and 33.4% glycerol removed by A. oryzae / C. utilis. • 18.6 g L−1 biomass, 1.7 U mL−1 amylase and 4.5 g L−1 ethanol produced. • Cultivation liquid used as 200 mL L−1 for high solids corn mash production. • Ethanol yield and production rate greatly improved by fermented backset use. [ABSTRACT FROM AUTHOR]

In order to improve the slow ethanol fermentation during acetic acid fermentation process of black raspberry vinegar (BRV), the microbiological and physicochemical aspects of the effects of indigenous Saccharomyces cerevisiae JBCC-21A were examined. The selected S. cerevisiae JBCC-21A showed better growth and ethanol production rates than the commercial yeast strains. The ethanol production rate was 3-times faster than the traditional method. Acetic acid fermentation by S. cerevisiae JBCC-21A began 10 days earlier than the traditional method and reached up to 60 g/L acetic acid. Bacterial counts revealed Acetobacter pasteurianus was the only dominant species throughout the inoculated acetic acid fermentation. The physicochemical and functional properties of the fermented vinegar using indigenous S. cerevisiae JBCC-21A maintained a high quality similar to the traditional method, while being the faster fermentation process. Thus, it is suggested that inoculation of the indigenous S. cerevisiae strain in order to shorten the fermentation time without affecting the quality of traditional BRV. [ABSTRACT FROM AUTHOR]

Graphical abstract Highlights • Controlling extracellular redox potential enhances the tolerance of yeast to furfural. • Intracellular redox status shows a tight correlation with extracellular redox potential. • −100 mV is an optimal redox level to obtain high biomass and ethanol titer. Abstract Furfural is a major toxic byproduct present in the hydrolysate of lignocellulosic biomass, which inhibits the growth and ethanol fermentation of Saccharomyces cerevisiae. To enhance yeast tolerance to furfural, extracellular redox potential (ORP) regulation was employed for the fermentation system through controlling ORP at −150 mV, −100 mV and −50 mV, respectively, by adjusting its aeration. When ORP was controlled at −100 mV, yeast cells exhibited improved growth, furfural degradation and ethanol production. Analysis of intracellular redox pairs such as NADH/NAD+ and GSH/GSSG indicated a correlation between extracellular ORP and intracellular redox homeostasis. Moreover, intracellular reactive oxygen species (ROS) caused by furfural stress decreased under the redox control condition, which consequently conferred yeast cells tolerance to furfural. Comparative transcriptome analysis for yeast cells sampling under the ORP control at −100 mV further revealed that the expression of genes for proliferation was up-regulated to degrade furfural more efficiently. These results demonstrated that extracellular ORP regulation would be a strategy for enhancing yeast tolerance to furfural stress during cellulosic ethanol fermentation. [ABSTRACT FROM AUTHOR]

Abstract Methods for obtaining high-concentration (ca. 40 g/L) ethanol from cassava stem (CS) were explored by investigating the effects of acid hydrolysis conditions, hydrolysate concentration, and intermittent inoculation of yeast on ethanol fermentation. Two-step acid hydrolysis demonstrated that a reduction in autoclaving temperature from 121 °C to 111 °C remarkably lowered the furfural concentration in the hydrolysate, with only ±10% differences in the glucose and xylose concentrations. For the concentration of sugars in the CS hydrolysate, the acid hydrolysate prepared with CS concentration of 200 g/L was used instead of distilled water for the second step of acid hydrolysis using new CS biomass (i.e., two-times acid hydrolysis). This produced a concentrated hydrolysate containing 95 g/L of glucose and 0.95 g/L furfural. Saccharomyces cerevisiae strain IAM 4178 was inoculated into the concentrated CS hydrolysate after verifying its furfural tolerance. Furfural was rapidly degraded and glucose was converted to ethanol during the startup period, whereas cell density decreased to approximately 10% of the initial value and ethanol production ceased by 72 h. IAM 4178 was then re-inoculated intermittently to maintain the cell density at 108 cells/mL, which eventually resulted in the complete uptake of glucose and the production of high concentrations of ethanol, up to 37.5 g/L. Highlights • Two-time acid hydrolysis of cassava stem to produce high-concentration bioethanol. • Reduction of furfural in the hydrolysate by lowering the autoclaving temperature. • Concentration of glucose in the hydrolysate by two-time acid hydrolysis. • Maintenance of yeast cell density by intermittent yeast inoculation. • Complete uptake of glucose and production of 37.5 g/L of ethanol. [ABSTRACT FROM AUTHOR]

Cassava starch is an abundant feedstock for biological transformation to ethanol, however, its industrial processing needs further improvements to enhance efficiency and cost-effectiveness. In the present study, a low-cost catalyst (CC-SO3H) was synthesized by partial carbonization and sulfonation of crystalline cellulose, which was thermally stable and reactive at 160 °C in 5 times repeated batch of thermohydrolysis of cassava starch. The catalyst was studied for its potential role in the hydrolysis of cassava starch as a standard feedstock. It was shown that the milling of cassava starch in the presence of the CC-SO3H catalyst improved the solid-state reaction that enhanced porosity, increased surface area and decreased crystallinity of the starch granules. These phenomena caused the rapid thermohydrolysis of starch with an exceptionally high starch conversion rate (96.43%), glucose yield (93.12%), and glucose selectivity (95.32%) within 2 h of reaction at 160 "C, 10 bar. The highest ethanol yield (0.43 g ethanol/g total reducing sugars) was achieved at 96 h of fermentation corresponding to the highest ethanol concentration of 15.41 g/L from the fermentation of hydrolysate of mixed-milling/thermo-hydrolysis at 160 C for 2 h of cassava starch. In addition, the reaction kinetics showed the feasibility of this process for robust bioethanol production from starchy feedstocks. [ABSTRACT FROM AUTHOR]

Environmental and energy concerns have increased interest in renewable energy sources, particularly biofuels. Thus the fermentation of glucose from sulfuric acid-hydrolyzed corn stover for the production of bioethanol has been explored using a combined acid retardation and continuous-effect membrane distillation treatment process. This process resulted in the separation of the sugars and acids from the acid-catalyzed hydrolysate, the removal of most of the fermentation inhibitors from the hydrolysate and the concentration of the detoxified hydrolysate. The recovery rate of glucose from the sugar-acid mixture using acid retardation was greater than 99.12% and the sulfuric acid was completely recovered from the hydrolysate. When the treated corn stover hydrolysate, containing 100 g/L glucose, was used as a carbon source, 43.06 g/L of ethanol was produced with a productivity of 1.79 g/(L∙h) and a yield of 86.31%. In the control experiment, where glucose was used as the carbon source these values were 1.97 g/(L∙h) and 93.10% respectively. Thus the integration of acid retardation and a continuous-effect membrane distillation process are effective for the production of fuel ethanol from corn stover. [ABSTRACT FROM AUTHOR]

In this study, the effectiveness of the ethanol fermentation pretreatment for biomethanation was examined. A sequential batch biomethanation experiment was performed for 130 days using artificial food waste, which was saccharized and ethanol fermented without any evaporation of ethanol. This is compared with a control experiment without pretreatment. Methane yields of the control and pretreatment series were 420 and 460 mL/g of the added volatile solids (VS) with 53% and 68% of methane content, 0.082 and 0.014 g of volatile suspended solids/g VS added for sludge yield, and 87% and 94% for VS biodegradability, respectively. In terms of material balance, the generation of carbon dioxide during biomethanation from an ethanol fermented substrate is lower in volume than that without it, because carbon dioxide is released during ethanol fermentation. The lower sludge generation of the ethanol fermented substrate results from the biomass yield of ethanol and free energy obtained from the biomethanation of ethanol, which is lower than that obtained from the biomethanation of starch. [ABSTRACT FROM AUTHOR]

Direct hydrolysis of a towel to glucose was investigated using steam explosion and microwave-assisted treatment to find effective uses for cotton waste. The maximum glucose yield by direct hydrolysis (based on an untreated towel) was 18.8%, obtained at a steam pressure of 5.5 MPa (at 271 °C) and steaming time of 5 min using steam explosion. For the microwave-assisted treatment, with a 1.0 (w/w)% sulfuric acid catalyst, the maximum glucose yield by direct hydrolysis was 28.9%, obtained at a microwave heating temperature of 200 °C for 7 min. The maximum total glucose yield (from both direct hydrolysis and enzymatic hydrolysis of treated residue) was 78.0%, attained at a microwave heating temperature of 200 °C for 7 min with 0.5 (w/w)% sulfuric acid catalyst. Furthermore, the maximum total glucose and valuable water soluble chemicals (cellobiose, 5-hydroxymethylfurfural, formic acid, and levulinic acid), 94.1%, were achieved at heating temperature of 200 °C for 10 min with 0.5 (w/w)% sulfuric acid catalyst. Finally, ethanol, 84.5% of conversion rate, could be produced using supernatant (it contained glucose) and microwave treated residue (200 °C for 7 min with 0.25 (w/w)% sulfuric acid catalyst) as carbon source for Saccharomyces cerevisiae with less fermentation inhibition. [ABSTRACT FROM AUTHOR]

Highlights • Ratio of α-amylase to glucoamylase is important in saccharifying cassava pulp (CP). • Cold hydrolysis at 50 °C followed by 30 °C is effective in CP saccharification. • A model was developed for predicting the performance of cold hydrolysis process. • A sequential process for bioconversion of CP to ethanol was proposed. • The proposed process yielded 27.4 g-ethanol/L, with 57.8% fermentation efficiency. Abstract The present study investigated cold hydrolysis of cassava pulp (CP) and the use of cold hydrolysis with simultaneous saccharification and fermentation (SSF) for ethanol production. Cold hydrolysis of 100 g-CP/L at 50 °C for 2 h, followed by at 30 °C for 72 h resulted in the production of 71.5 ± 1.8 g/L of reducing sugar, with a yield of 0.72 g/g-CP. A mathematical model describing the cold hydrolysis process was subsequently developed. The model proved to be applicable for other cold hydrolysis systems with satisfactory results. The sequential process of cold hydrolysis at 50 °C for 2 h, followed by SSF at 30 °C for 72 h gave 27.4 g-ethanol/L, with a productivity of 0.37 g/(L h) and a fermentation efficiency of 57.58%. Based on the results, a bioconversion process for CP to ethanol was proposed. In this process, 1 kg of ethanol could be produced from 3.65 kg of CP without any nutrient supplementation. [ABSTRACT FROM AUTHOR]

Highlights • Lignocellulosic waste can serve as a microbial carrier in bioconversion processes. • Lignocellulosic carriers were hydrothermally modified for microbial immobilization. • Specific surface area and porosity of corn stalks were improved by the modification. • Conditions of 200 °C for 20 min with a 1 cm size were optimal for modifying the carriers. • Modified carriers showed better immobilization and ethanol fermentation performance. Abstract Microbial immobilization is a key approach to enhance fermentation performance, especially with the advantages of low costs and the good surface properties of the lignocellulosic wastes, which can serve as natural carriers. This study investigated the capability of hydrothermal modification to improve the physicochemical properties of corn stalk carriers for microbial immobilization. Hydrothermal modification can degrade lignocellulosic structure by dissolving part of the components at different conditions. A temperature of 200 °C for 20 min with a carrier side length of 1 cm were selected as the optimal modification conditions. With these conditions, the specific surface area and porosity of the modified corn stalk carriers were 2.44 m2/g and 0.011 cm3/g, respectively, which were twice the values of untreated carriers. Immobilization tests showed that bacteria and yeast grew stably inside the modified carriers and that the immobilized biomass was 49% greater than that of the untreated carriers. After fermentation with bacteria, ethanol production in the modified carrier system was increased by 57% and 99% compared to the untreated carrier system and free bacteria, respectively. These findings confirmed that hydrothermal modification is efficient to improve lignocellulosic carrier properties to achieve better immobilization and fermentation performance, thus benefiting to the utilization of lignocellulosic wastes and their bioconversion processes. [ABSTRACT FROM AUTHOR]

Highlights • A continuous fermentation process with a delayed product inhibition is considered. • Stability analysis of the control system with PI algorithm and process delay is given. • The negative effect of measurement noises is considered. • The process is stabilized for any delay and for a sufficiently high controller gain. • Filtering of the process variable reduces the effect of noises, but affects stability. Abstract The paper addresses the problem of stabilization of ethanol concentration in continuous fermentation processes. The stability analysis is performed for a class of continuous fermentation processes assuming that the possible oscillatory behavior results from a delayed response of microorganisms to inhibitory effects of ethanol concentration and that the specific growth rate is a monotonic function of substrate and a monotonic function of product concentrations. Treating dilution rate as the manipulated variable, it is shown that the classical PI controller is able to stabilize the oscillating process in the whole range of operating regions. Moreover, for a sufficiently large controller gain, the closed-loop system is asymptotically stable for any process delay. However, a high-gain controller may lead to extensive control signal activity and wear of actuator in the presence of measurement noises. To reduce the negative effect of measurement noises, the measured process variable is filtered by using the first order filter. Then, it is shown that the closed-loop system with the filter can also be stabilized for any process delay. The simulation results confirmed the effectiveness of the proposed closed-loop strategy with and without filtering. [ABSTRACT FROM AUTHOR]

Highlights • Alkali and acid pretreated biomass slurries were mixed for ethanol production. • This approach saved 40–50% alkali/acid use. • This approach improved fermentability. Abstract Alkali and acid pretreatment based processes for lignocellulosic biofuel production typically consume substantial amount of alkali and acid. In addition, alkali pretreatment generates compounds with high toxicity rendering washing/detoxification inevitable. In this work, alkali (AL) and dilute acid (DA) pretreatment based processes were compared side by side for ethanol production from corn stover (CS) using commercial enzymes and Zymomonas mobilis 8b strain. Next, AL and DA pretreated CS slurries (without removing liquid stream) were mixed to neutralize for enzymatic hydrolysis and ethanol fermentation. The digestibility and fermentability of the mixed pretreated biomass were systematically studied and compared to unmixed pretreated biomass. Results showed that mixed biomass process saved 40–50% alkali/acid use and enhanced fermentability compared to AL-CS. This work suggests that it could be beneficial to have both alkali and acid pretreatments operating in a biorefinery, which could accommodate various feedstocks. [ABSTRACT FROM AUTHOR]

Lignocellulosic materials are good feedstocks for ethanol production. Mathematical models give information about kinetic-metabolic nature of fermentation. The objectives of this study were to determine the chemical composition of non-detoxified tea processing waste hydrolysate (ND-TPWH) and detoxified tea processing waste hydrolysate (D-TPWH), to generate the ethanol from the D-TPWH, and to model the ethanol fermentation in D-TPWH using models including modified Gompertz, re-modified Gompertz, modified logistic, re-modified logistic, modified Richards, re-modified Richards, Stannard, Baranyi, Weibull, and Morgan-Mercer-Flodin. Results indicated that 11.91% of d -glucuronic acid, 7.28% of acetic acid, 98.12% of hydroxymethylfurfural, and 76.88% of phenolics were adsorbed by detoxification process. Ethanol yields by Saccharomyces cerevisiae and Scheffersomyces stipitis (ATCC 58784 and ATCC 58785) were 35.9, 38.98, and 33.87%, respectively. Regarding modeling, depending on the model comparison results including root-mean-square-error, mean-absolute-error, and regression coefficient, the experimental data of ethanol production and sugar consumption were successfully forecasted using Baranyi and Weibull models for S. cerevisiae ; using Morgan-Mercer-Flodin model for S. stipitis (ATCC 58784); and using Stannard model for S. stipitis (ATCC 58785), respectively. Consequently, this was the first report on the ethanol production from D-TPWH and its modeling. TPW can be a good feedstock for ethanol production by the xylose-fermenting yeasts. Suitable flexible models could be applied for more progress of ethanol production process in D-TPWH. [ABSTRACT FROM AUTHOR]

The development of sustainable bioethanol fuel production from food waste has increasingly become an attractive topic. Food waste is recognized as the most available and costless feedstock. Therefore, ethanol production has been adopted as cost-efficient and an ecological way for FW disposal. This paper reviewed the microorganisms utilized for ethanol fermentation, the effect of enzymatic hydrolysis on ethanol concentration, optimization of accurate process parameters, and recycling of huge volumes of stillage for ethanol production towards reducing any incurred environmental burdens and minimizing the cost. The statistical tools which may enhance the process efficiency had been presented. Also, the perspective and the future development were introduced. All these aimed to fully utilize the food waste and also reduce the cost for side-product in this process; proper operation conditions and the control methods for stillage recycling were considered as the methods to improve ethanol fermentation from food waste. [ABSTRACT FROM AUTHOR]

Graphical abstract Highlights • Content of higher alcohols is closely related to the type of raw material used. • Trace-amount alcohols are mainly associated with method of starch liberation. • Initial pH of sweet mashes has an effect on the content of 1- and 2-propanol. Abstract The qualitative and quantitative profile of volatile compounds in agricultural distillate depends on numerous factors. Many such compounds, including higher alcohols, occur naturally, due to the metabolism of yeast during ethanol fermentation. Some are difficult to remove in the rectification process. For these reasons, it is desirable to reduce the synthesis of volatile compounds, among others higher alcohols, during the fermentation process. The results of the presented study showed that the raw materials, the method of starch liberation, and the initial pH of the sweet mashes affected the concentrations of higher alcohols in the distillates. Higher concentrations of 1-butanol, 1-propanol, and 2-propanol were found in distillates obtained from mashes prepared by the thermal-pressure method, while the remaining compounds occurred at higher concentrations in distillates obtained using the pressureless starch liberation method. The type of raw materials used had an influence on the concentration of the majority of higher alcohols. The content of 2-propanol, 2-methyl-1-propanol, 1-propanol, 1-butanol, 3-methyl-1-butanol, and 2-phenylethanol was lower in distillates obtained from mashes with an initial pH of 4, in comparison with those obtained from mashes with the pH set at 5.5. [ABSTRACT FROM AUTHOR]

Abstract: Dekkera bruxellensis is continuously changing its status in fermentation processes, ranging from a contaminant or spoiling yeast to a microorganism with potential to produce metabolites of biotechnological interest. In spite of that, several major aspects of its physiology are still poorly understood. As an acetogenic yeast, minimal oxygen concentrations are able to drive glucose assimilation to oxidative metabolism, in order to produce biomass and acetate, with consequent low yield in ethanol. In the present study, we used disulfiram to inhibit acetaldehyde dehydrogenase activity to evaluate the influence of cytosolic acetate on cell metabolism. D. bruxellensis was more tolerant to disulfiram than Saccharomyces cerevisiae and the use of different carbon sources revealed that the former yeast might be able to export acetate (or acetyl‐CoA) from mitochondria to cytoplasm. Fermentation assays showed that acetaldehyde dehydrogenase inhibition re‐oriented yeast central metabolism to increase ethanol production and decrease biomass formation. However, glucose uptake was reduced, which ultimately represents economical loss to the fermentation process. This might be the major challenge for future metabolic engineering enterprises on this yeast. [ABSTRACT FROM AUTHOR]

Maize starch was irradiated by a Co60 irradiator with different doses. The morphology and physicochemical properties of native and irradiated starches were investigated. Scanning electron microscopy showed that the shape and size of starch granules did not change after irradiation. However, the irradiated starch granules were easily destroyed by dissolution. Irradiation also caused the change of starch color, the decrease in the pH value, light transmittance, stability index, degree of polymerization, total sugar content, and the increase in the swelling index and the reducing sugar content. In this study, irradiated maize starch was also used as material for ethanol fermentation to investigate its potential as a pretreatment method. Results showed that the ethanol yield of cooked and raw starch fermentation using irradiated starch increased by 20.41 % and 5.18 %, respectively, and the ethanol concentration increased by 3 % and 2 %. This finding indicated that irradiation effectively improved the utilization rate of maize starch, making it an effective pretreatment method for ethanol fermentation. [ABSTRACT FROM AUTHOR]

We demonstrate a technique that uses microdroplets for culturing and selecting bacterial cultures in a model biotechnological application. We propose an assay for determination of ethanol concentration that provides increased dynamic range and is compatible with droplet microfluidic screening technologies. The assay comprises two enzymes—alcohol oxidase (AOX) and horseradish peroxidase (HRP)—and a colorimetric readout system of phenol-4-sulfonic acid (PSA) and 4-aminoantipyrine (4-AAP). The microdroplet method provides high repeatability (a relative error of measured ethanol concentration < 5%), high specificity for ethanol, low consumption of reagents and wide dynamic range (1–70 g·L-1) compared to existing assays. We report the use of this method in a screen of ethanol generation efficiency of Zymomonas mobilis (strain 3881) against the concentration of glucose in the culture media.