Ethanol Production Research Articles

Mix-method toolbox for monitoring greenhouse gas production and microbiome responses to soil amendments

In this study, we adopt an interdisciplinary approach, integrating agronomic field experiments with soil chemistry, molecular biology techniques, and statistics to investigate the impact of organic residue amendments, such as vinasse (a by-product of sugarcane ethanol production), on soil microbiome and greenhouse gas (GHG) production. The research investigates the effects of distinct disturbances, including organic residue application alone or combined with inorganic N fertilizer on the environment. The methods assess soil microbiome dynamics (composition and function), GHG emissions, and plant productivity. Detailed steps for field experimental setup, soil sampling, soil chemical analyses, determination of bacterial and fungal community diversity, quantification of genes related to nitrification and denitrification pathways, measurement and analysis of gas fluxes (N2O, CH4, and CO2), and determination of plant productivity are provided. The outcomes of the methods are detailed in our publications (Lourenço et al., 2018a; Lourenço et al., 2018b; Lourenço et al., 2019; Lourenço et al., 2020). Additionally, the statistical methods and scripts used for analyzing large datasets are outlined. The aim is to assist researchers by addressing common challenges in large-scale field experiments, offering practical recommendations to avoid common pitfalls, and proposing potential analyses, thereby encouraging collaboration among diverse research groups.•Interdisciplinary methods and scientific questions allow for exploring broader interconnected environmental problems.•The proposed method can serve as a model and protocol for evaluating the impact of soil amendments on soil microbiome, GHG emissions, and plant productivity, promoting more sustainable management practices.•Time-series data can offer detailed insights into specific ecosystems, particularly concerning soil microbiota (taxonomy and functions).

Which bioenergy with carbon capture and storage (BECCS) pathways can provide net-negative emissions?

Countries are considering different options to achieve net zero emissions including Bioenergy with carbon capture and storage (BECCS), which is the process of capturing and storing CO2 from processes that use bioenergy to produce heat, electricity, or biofuels. However, this technology faces sustainability concerns and possesses complex value chains of its emissions. Adding further to this complexity, the literature indicates two opposing views with respect to the potential of BECCS in terms of being able (or unable) to achieve negative emissions. Hence, this paper analyzes in detail a wide range of BECCS pathways in terms of their ability to achieve negative emissions along with their associated costs. Out of seven assessed pathways, our analysis shows that corn to ethanol and biomethane production from maize BECCS pathway in the USA, biomethane production from wet manure in Europe, and baling of straw pellets with trans-Atlantic shipment can achieve negative emissions at a cost of 50, 108, 159, and 232 dollars per ton of CO2 ($/tCO2) respectively. Other technologies like poplar pellets, forest residue, and agricultural residue with trans-Atlantic shipment are not able to achieve negative emissions.

The Macroalga Kappaphycus alvarezii as a Potential Raw Material for Fermentation Processes within the Biorefinery Concept: Challenges and Perspectives

Seaweed is a fast-growing biomass source that is currently studied as feedstock for sustainable industrial production in a wide variety of markets. Being composed mostly of polysaccharides, macroalgae can be integrated in biorefineries for obtaining bioproducts via fermentation. Kappaphycus alvarezii has been introduced experimentally to Brazil’s south coastline in 1995 and is now cultivated on a large scale to keep up with the high carrageenan demand in various industrial sectors. In this review article, an introduction is given on renewable biomass and environmental issues, focusing especially on third-generation biomass and its promising features and use advantages. Later on, the processing of K. alvarezii for the use of its saccharide portion for fermentative processes is approached. The current state of research conducted alongside challenges and hurdles in K. alvarezii hydrolysate fermentation processes provides insight into future studies needed to make new fermentation processes viable. Next, some fermentation products are discussed, and the metabolism of galactose in microorganisms is also presented to bring to light other possible fermentation products that are not yet, but can be, obtained from K. alvarezii. Finally, a simple and comprehensive scheme for K. alvarezii fermentation biorefinery is presented to demonstrate a generic example for a possible configuration for obtaining valuable bio-products. In the literature, production of ethanol and lactic acid were already reported from K. alvarezii. This review aims to help envision new industrial processes that can be developed for this most valuable macroalga.

Isolation, identification, and tolerance analysis of yeast during the natural fermentation process of Sidamo coffee beans.

Yeast, which plays a pivotal role in the brewing, food, and medical industries, exhibits a close relationship with human beings. In this study, we isolated and purified 60 yeast strains from the natural fermentation broth of Sidamo coffee beans to screen for indigenous beneficial yeasts. Among them, 25 strains were obtained through morphological characterization on nutritional agar medium from Wallerstein Laboratory (WL), with molecular biology identifying Saccharomyces cerevisiae strain YBB-47 and the remaining 24 yeast strains identified as Pichia kudriavzevii. We investigated the fermentation performance, alcohol tolerance, SO2 tolerance, pH tolerance, sugar tolerance, temperature tolerance, ester production capacity, ethanol production capacity, H2S production capacity, and other brewing characteristics of YBB-33 and YBB-47. The results demonstrated that both strains could tolerate up to 3% alcohol by volume at a high sucrose mass concentration (400g/L) under elevated temperature conditions (40 ℃), while also exhibiting a remarkable ability to withstand an SO2 mass concentration of 300g/L at pH 3.2. Moreover, S. cerevisiae YBB-47 displayed a rapid gas production rate and strong ethanol productivity. whereas P. kudriavzevii YBB-33 exhibited excellent alcohol tolerance. Furthermore, this systematic classification and characterization of coffee bean yeast strains from the Sidamo region can potentially uncover additional yeasts that offer high-quality resources for industrial-scale coffee bean production.

Evaluation of sugar content and bioethanol production of Ethiopian local varieties “Nech Tinkish” and “Hawaye” sweet sorghum (Sorghum bicolor (L.))

Diversifying the use of climate-smart crops such as sweet sorghum has the potential to solve integrated food, bioenergy, feed, and land management problems. The study’s purpose is to quantify the sugar content of Nech Tinkish (v1) and Hawaye (v2) Ethiopian sweet sorghum varieties and investigate the interaction effect of fermentation parameters to determine their capacity for ethanol production. Sweet sorghum varieties were analyzed to determine their difference in °Brix content by extracting their juices. The juice was clarified using milk lime. Its total soluble sugars, total carbohydrates, and reducing sugars were determined using a digital refractometer, phenol sulfuric acid, and 3,5-dinitrosalicylic acid, respectively. A completely randomized factorial was employed to evaluate ethanol production capacity, and the ethanol content was estimated using a potassium dichromate solution. The °Brix results revealed that v2 had a higher sugar concentration than v1. Additionally, the estimated carbohydrate content of the juice ranged from 37.402 to 157.641 g/L. The estimated reducing sugar also varied from 4.644 to 33.412 g/L. Therefore, the estimated reducing sugar showed the hydrolysis of sweet sorghum juice by invertase and sulfuric acid produced more fermentable sugars. Fermentation at 30 °C with pH 4.5 incubated for 4 days yields the highest ethanol, and v2 yields higher (15.31%) ethanol, compared to v1 produced 14.85%. This study showed a basis for the existence of two sugar-rich climate smart sweet sorghum varieties with an extraordinary amount of sugar used as a source of biofuel and food simultaneously in a single plot of land.

Digestibility and nutritional parameters of maize ethanol coproducts and xylanase for broiler diets

ABSTRACT 1. The objective of this study was to determine the nutritional and energy values of four maize distiller’s dried grains with solubles (DDGS) and one maize high protein distiller’s dried grains (HP-DDG) from ethanol production plants in Brazil; to evaluate the digestibility, performance, nitrogen balance and energy values for broiler chickens fed diets containing these coproducts (Experiment I); and to evaluate the effects of xylanase inclusion in diets containing maize DDGS for broilers on energy availability, digestibility, nitrogen balance and gastrointestinal morphometry (Experiment II). 2. For each experiment, 180 broiler chickens aged 17 and 30 days with initial weights of 450 ± 18 g and 1228 ± 33 g, respectively, were used; the chickens were distributed into 36 metabolism cages. The experimental design consisted of complete randomised blocks, with six replications per treatment and five birds per experimental unit. The treatments consisted of a basal diet (BD) and five test diets containing maize ethanol coproducts (Experiment I) one BD and five test diets containing DDGS with inclusions of 0, 8,000, 16,000, 24,000 and 32,000 BXU/kg xylanase (Experiment II). In Experiment I, HP-DDG and DDGS2 presented higher AME and AMEn values (14.1 and 13.9 MJ/kg and 13.4 and 13.3 MJ/kg, respectively), than did the other coproducts (p < 0.05). Compared with DDGS1 and DDGS3, DDGS4 and HP-DDG had higher digestible CP values (p < 0.05). In Experiment II, the inclusion of the enzyme quadratically affected the values of digestible CP and digestible EE (p < 0.05), with the maximum values occurring with the inclusion of 18 750 and 22,170 BXU/kg of xylanase, respectively. 3. The digestible NDF and digestible MM values linearly increased with the inclusion of xylanase (p < 0.05). The addition of xylanase had no effect on gastrointestinal morphometry (p > 0.05). It was concluded that the inclusion of between 18,000 and 22,000 BXU/kg of xylanase resulted in better digestible CP and digestible EE values.

Economic Research on Ethanol Feed-Use Coproducts: A Review, Synthesis, and Path Forward

During the mid-2000s to the early 2010s, the domestic ethanol industry witnessed substantial growth, with ethanol coproducts emerging as vital elements for plant profitability and livestock feeding. Initially serving as supplementary revenue streams, coproducts from ethanol production have evolved into diverse value-added offerings, bolstering revenue streams, and sustaining profit margins. This study reviews existing economic research on ethanol coproducts, detailing methodologies, product focus, and research locations. Initially gathering 972 articles from 9 databases, 110 articles were synthesized. We find that most studies primarily examined the growth and future of the ethanol industry with a limited focus on specific coproducts. Feed-use distillers’ grains, especially dried distillers’ grains, were the most widely published while newer coproducts like pelletized, de-oiled, and high-protein distillers’ grains were relatively understudied. Non-feed-use products were notably overlooked, highlighting the need for exploration beyond conventional applications. The evolving market landscape for ethanol co-products has surpassed published academic understanding of the economic tradeoffs necessitating further research into product dynamics, pricing, marketing, market structures, and regulatory frameworks. This highlights and underscores the importance of investigating value-added grains across diverse commodities and geographic contexts to inform strategic decision-making and policy formulation.

A new Zymomonas mobilis platform strain for the efficient production of chemicals

BackgroundZymomonas mobilis is well known for its outstanding ability to produce ethanol with both high specific productivity and with high yield close to the theoretical maximum. The key enzyme in the ethanol production pathway is the pyruvate decarboxylase (PDC) which is converting pyruvate to acetaldehyde. Since it is widely considered that its gene pdc is essential, metabolic engineering strategies aiming to produce other compounds derived from pyruvate need to find ways to reduce PDC activity.ResultsHere, we present a new platform strain (sGB027) of Z. mobilis in which the native promoter of pdc was replaced with the IPTG-inducible PT7A1, allowing for a controllable expression of pdc. Expression of lactate dehydrogenase from E. coli in sGB027 allowed the production of D-lactate with, to the best of our knowledge, the highest reported specific productivity of any microbial lactate producer as well as with the highest reported lactate yield for Z. mobilis so far. Additionally, by expressing the L-alanine dehydrogenase of Geobacillus stearothermophilus in sGB027 we produced L-alanine, further demonstrating the potential of sGB027 as a base for the production of compounds other than ethanol.ConclusionWe demonstrated that our new platform strain can be an excellent starting point for the efficient production of various compounds derived from pyruvate with Z. mobilis and can thus enhance the establishment of this organism as a workhorse for biotechnological production processes.

Methane Production from Sugarcane Vinasse Biodigestion: An Efficient Bioenergy and Environmental Solution for the State of São Paulo, Brazil

This study mapped the bioenergy production from sugarcane vinasse according to the mesoregions of the State of São Paulo (SP), Brazil, assessing the magnitude of biogas-derived electricity and biomethane production and estimating the greenhouse gas (GHG) emissions. SP holds 45% of the Brazilian ethanol-producing plants, in which 1.4 million m3 of carbon-rich vinasse are generated daily. The electricity generated from vinasse has the potential to fully supply the residential consumption (ca. 6.5 million inhabitants) in the main sugarcane-producing mesoregions of the state (Ribeirão Preto, São José do Rio Preto, Bauru, Araçatuba and Presidente Prudente). In another approach, biomethane could displace almost 3.5 billion liters of diesel, which represents a 26% abatement in the annual state diesel consumption. Energetically exploiting biogas is mandatory to prevent GHG-related drawbacks, as the eventual emission of methane produced under controlled conditions (261.2 × 106 kg-CO2eq d−1) is ca. 7-fold higher than the total emissions estimated for the entire ethanol production chain. Meanwhile, replacing diesel with biomethane can avoid the emission of 45.4 × 106 kg-CO2eq d−1. Implementing an efficient model of energy recovery from vinasse in SP has great potential to serve as a basis for expanding the utilization of this wastewater in Brazil.

Optimizing Bioethanol (C2H5OH) Yield of Sweet Sorghum Varieties in a Semi-Arid Environment: The Impact of Deheading and Deficit Irrigation

Bioethanol production offers promise in mitigating environmental impacts from ethanol consumption despite water scarcity. This study endeavors to evaluate the nuanced influence of different deheading times (45 days before harvest, 21 days before harvest, and no deheading) along with varying water regimes on select sweet sorghum cultivars (Honey, Willy, MN1500, and Atlas), focusing on yield traits, theoretical ethanol production, and water productivity. Findings underscore the substantial impact of cultivation practices on bioethanol yield. A water deficit ranging from 30% to 70% resulted in a discernible reduction in stalk yields of 17.86% to 18.54% and in sugar yields of 0.2 to 0.31 Mg ha−1, accompanied by a corresponding decline in theoretical ethanol yield of 120.9 to 180.9 L ha−1. Additionally, notable enhancements in Brix and sugar content of 16.32% to 18.42% and 16.81% to 19.03%, respectively, were observed across both seasons. Of particular significance, the Honey variety, subjected to a 30% water deficit and deheading at 21 days before harvest, demonstrated exceptional growth and yield characteristics. These empirical insights furnish valuable guidance for optimizing sweet sorghum cultivation practices, thereby augmenting sustainable bioethanol production and propelling forward the frontier of renewable energy technologies towards a more environmentally sustainable future.

Fermentation of Rice Straw and Its Hydrolysate with Sludge for Ethanol Production

Fermentation of Rice Straw and Its Hydrolysate with Sludge for Ethanol Production

Mo4+-Doped CuS Nanosheet-Assembled Hollow Spheres for CO2 Electroreduction to Ethanol in a Flow Cell.

Electrocatalytic CO2 reduction reaction (CO2RR) to ethanol has been widely researched for potential commercial application. However, it still faces limited selectivity at a large current density. Herein, Mo4+-doped CuS nanosheet-assembled hollow spheres are constructed to address this issue. Mo4+ ion doping modifies the local electronic environments and diversifies the binding sites of CuS, which increases the coverage of linear *COL and produces bridge *COB for subsequent *COL-*COH coupling toward ethanol production. The optimal Mo9.0%-CuS can electrocatalyze CO2 to ethanol with a faradaic efficiency of 67.5% and a partial current density of 186.5 mA cm-2 at -0.6 V in a flow cell. This work clarifies that doping high valence transition metal ions into Cu-based sulfides can regulate the coverage and configuration of related intermediates for ethanol production during the CO2RR in a flow cell.

Boosting Solar‐Driven CO2 Conversion to Ethanol via Single‐Atom Catalyst with Defected Low‐Coordination Cu‐N2 Motif

In this paper, we report a high‐performance carbon nitride supported Cu single‐atom catalyst featuring defected low‐coordination Cu‐N2 motif (Cu‐N2‐V). Lead many recently reported photocatalysts and its Cu‐N3 and Cu‐N4 counterparts, Cu‐N2‐V exhibits superior photocatalytic activity for CO2 reduction to ethanol, delivering 69.8 μmol g‐1 h‐1 ethanol production rate, 97.8% electron‐based ethanol selectivity, and a yield of ~10 times higher than Cu‐N3 and Cu‐N4. Revealed by the extensive experimental investigation combined with the DFT calculation, the superior photoactivity of Cu‐N2‐V stems from its unique defected Cu‐N2 configuration. Firstly, Cu in Cu‐N2‐V exist in Cu+/Cu2+ dual valence states, although predominantly in Cu+. The Cu+ sites support CO2 activation and the Cu+/Cu2+ sites are conducive for strong *CO adsorption and subsequent *CO‐*CO dimerization enabling C‐C coupling. Secondly, the Cu sites in Cu‐N2‐V are rich in electrons and thus highly active. Together they dictate the rate‐determining step on CO2 photoreduction to ethanol and lower the Gibbs free energy change. Furthermore, the defected configuration also promotes light adsorption and charge separation efficiency. Collectively, these make Cu‐N2‐V an effective and high‐performance catalyst for solar‐driven CO2 conversion to ethanol. This study also reveals the valence state change of Cu in Cu‐N2‐V during the CO2 photoreduction reaction.

Boosting Solar-Driven CO2 Conversion to Ethanol via Single-Atom Catalyst with Defected Low-Coordination Cu-N2 Motif.

In this paper,we report a high-performance carbon nitride supported Cu single-atom catalyst featuring defected low-coordination Cu-N2 motif (Cu-N2-V). Lead many recently reported photocatalysts and its Cu-N3 and Cu-N4 counterparts, Cu-N2-V exhibits superior photocatalytic activity for CO2 reduction to ethanol, delivering 69.8 μmol g-1 h-1 ethanol production rate, 97.8% electron-based ethanol selectivity, and a yield of ~10 times higher than Cu-N3 and Cu-N4. Revealed by the extensive experimental investigation combined with the DFT calculation, the superior photoactivity of Cu-N2-V stems from its unique defected Cu-N2 configuration. Firstly, Cu in Cu-N2-V exist in Cu+/Cu2+ dual valence states, although predominantly in Cu+. The Cu+ sites support CO2 activation and the Cu+/Cu2+ sites are conducive for strong *CO adsorption and subsequent *CO-*CO dimerization enabling C-C coupling. Secondly, the Cu sites in Cu-N2-V are rich in electrons and thus highly active. Together they dictate the rate-determining step on CO2 photoreduction to ethanol and lower the Gibbs free energy change. Furthermore, the defected configuration also promotes light adsorption and charge separation efficiency. Collectively, these make Cu-N2-V an effective and high-performance catalyst for solar-driven CO2 conversion to ethanol. This study also reveals the valence state change of Cu in Cu-N2-V during the CO2 photoreduction reaction.

Nematode community structure in sugarcane fields under continuous vinasse fertigation

Sugarcane vinasse is a residue from ethanol production, which has high content of nutrients and organic matter, improving chemical, physical and biological changes in the soil. This study evaluated nematode community structure, food web, metabolic footprint, and soil quality in sugarcane fields under continuous vinasse fertigation. Soil samples were collected from four sugarcane fields in Pernambuco State, Brazil, under continuous vinasse fertigation for nematode identification at the genus (plant parasites) and family (free-living) level, physical and chemical analyses of the soil. Nine genera of plant-parasitic nematodes and five families of free-living nematodes were identified in sugarcane fields. Plant-parasitic nematodes were dominant, except in the field with five years of continuous vinasse application. The genera Mesocriconema, Meloidogyne, Helicotylenchus and Pratylenchus were frequent in all fields. Predators and fungivores nematodes were few abundant, although, according to the soil food web, all sugarcane fields were low to moderately disturbed environments. A positive correlation was observed between the microbial respiration and Rhabdtidae in fields with five years of vinasse application, and between the electrical conductivity of the soil and Meloidogyne in fields without vinasse application. We highlight those alterations in the structure and composition of the nematode community, and soil characteristics that can occur due to vinasse fertigation in sugarcane fields.

Quantitative assessment of ash effects on densifying lignocellulosic biomass with chemicals followed by autoclave (DLCA) pretreatment

Ash in lignocellulose is critical in interfering biomass pretreatment. Densifying Lignocellulosic biomass with Chemicals followed by Autoclave (DLCA) is a novel and efficient pretreatment method, whereas the specific effects of ash on DLCA pretreatment remain unclear. Herein, we quantitatively investigated the impacts of ash contents on the pretreatment effectiveness, enzymatic digestibility, and fermentability of DLCA pretreated corn stover (DLCA-CS). With the ash contents increasing from 3.36 % to 9.50 %, the DLCA-CS with addition of sulfuric acid (sa) showed decreased pretreatment efficiency and enzymatic digestibility, whereas these characteristics of DLCA-CS with addition of calcium hydroxide (ch) were improved with increasing ash contents. Further pH measurement during the biomass pretreatment revealed that the anions involved in ash neutralized the sulfuric acid, thus decreasing the pretreatment efficiency and enzymatic digestibility of DLCA(sa)-CS, while the anions can cooperated with the calcium hydroxide, thus increasing the pretreatment efficiency and enzymatic digestibility of DLCA(ch)-CS. The highest ethanol production of DLCA(sa)-CS (60.7 g/L) and DLCA(ch)-CS (66.5 g/L) at 25 % solid loading was achieved with a low ash content of 3.36 % without detoxification step. This study highlights the importance of controlling the ash contents for enhancing the efficiency of DLCA pretreatment.

The time-varying dynamic impact of US renewable energy prices on agricultural prices in China: the case of fuel ethanol.

This paper intends to look into the time-varying dynamic impact of US fuel ethanol, one of the renewable energy sources, on the prices of agricultural products (specifically corn, soybeans, rice, and wheat) in China based on monthly price data from January 2000 to January 2023. To achieve this, a time-varying parameter vector autoregressive (TVP-VAR) model is employed, which takes into account structural changes in emergencies through time-varying parameters. The empirical results show that the equal-interval impulse responses of price fluctuations in agricultural commodities are primarily positive to variations in fuel ethanol prices and production. And the intensity and direction of the effects vary at distinct time lags. Additionally, the magnitude of these responses is most pronounced in the short term for all agricultural commodities except for corn, and the duration of the impulse responses at different time points is generally longer for corn prices compared to other commodities. The study also reveals that the influence of US fuel ethanol on Chinese agricultural commodity prices is not substantial on the whole. Therefore, there is a necessity to advance the growth of biofuels and provide policy support and financial subsidies for agricultural products earmarked for food production. These actions could shed insights into the progression of Chinese renewable energy and food policies, ensuring the stability of the market in the long run.

Optimal trade-off between boosted tolerance and growth fitness during adaptive evolution of yeast to ethanol shocks

BackgroundThe selection of Saccharomyces cerevisiae strains with higher alcohol tolerance can potentially increase the industrial production of ethanol fuel. However, the design of selection protocols to obtain bioethanol yeasts with higher alcohol tolerance poses the challenge of improving industrial strains that are already robust to high ethanol levels. Furthermore, yeasts subjected to mutagenesis and selection, or laboratory evolution, often present adaptation trade-offs wherein higher stress tolerance is attained at the expense of growth and fermentation performance. Although these undesirable side effects are often associated with acute selection regimes, the utility of using harsh ethanol treatments to obtain robust ethanologenic yeasts still has not been fully investigated.ResultsWe conducted an adaptive laboratory evolution by challenging four populations (P1–P4) of the Brazilian bioethanol yeast, Saccharomyces cerevisiae PE-2_H4, through 68–82 cycles of 2-h ethanol shocks (19–30% v/v) and outgrowths. Colonies isolated from the final evolved populations (P1c–P4c) were subjected to whole-genome sequencing, revealing mutations in genes enriched for the cAMP/PKA and trehalose degradation pathways. Fitness analyses of the isolated clones P1c–P3c and reverse-engineered strains demonstrated that mutations were primarily selected for cell viability under ethanol stress, at the cost of decreased growth rates in cultures with or without ethanol. Under this selection regime for stress survival, the population P4 evolved a protective snowflake phenotype resulting from BUD3 disruption. Despite marked adaptation trade-offs, the combination of reverse-engineered mutations cyr1A1474T/usv1Δ conferred 5.46% higher fitness than the parental PE-2_H4 for propagation in 8% (v/v) ethanol, with only a 1.07% fitness cost in a culture medium without alcohol. The cyr1A1474T/usv1Δ strain and evolved P1c displayed robust fermentations of sugarcane molasses using cell recycling and sulfuric acid treatments, mimicking Brazilian bioethanol production.ConclusionsOur study combined genomic, mutational, and fitness analyses to understand the genetic underpinnings of yeast evolution to ethanol shocks. Although fitness analyses revealed that most evolved mutations impose a cost for cell propagation, combination of key mutations cyr1A1474T/usv1Δ endowed yeasts with higher tolerance for growth in the presence of ethanol. Moreover, alleles selected for acute stress survival comprising the P1c genotype conferred stress tolerance and optimal performance under conditions simulating the Brazilian industrial ethanol production.

Key role of K+ and Ca2+ in high-yield ethanol production by S. Cerevisiae from concentrated sugarcane molasses

BackgroundSaccharomyces cerevisiae is an important microorganism in ethanol synthesis, and with sugarcane molasses as the feedstock, ethanol is being synthesized sustainably to meet growing demands. However, high-concentration ethanol fermentation based on high-concentration sugarcane molasses—which is needed for reduced energy consumption of ethanol distillation at industrial scale—is yet to be achieved.ResultsIn the present study, to identify the main limiting factors of this process, adaptive laboratory evolution and high-throughput screening (Py-Fe3+) based on ARTP (atmospheric and room-temperature plasma) mutagenesis were applied. We identified high osmotic pressure, high temperature, high alcohol levels, and high concentrations of K+, Ca2+, K+ and Ca2+ (K+&Ca2+), and sugarcane molasses as the main limiting factors. The robust S. cerevisiae strains of NGT-F1, NGW-F1, NGC-F1, NGK+, NGCa2+ NGK+&Ca2+-F1, and NGTM-F1 exhibited high tolerance to the respective limiting factor and exhibited increased yield. Subsequently, ethanol synthesis, cell morphology, comparative genomics, and gene ontology (GO) enrichment analysis were performed in a molasses broth containing 250 g/L total fermentable sugars (TFS). Additionally, S. cerevisiae NGTM-F1 was used with 250 g/L (TFS) sugarcane molasses to synthesize ethanol in a 5-L fermenter, giving a yield of 111.65 g/L, the conversion of sugar to alcohol reached 95.53%. It is the highest level of physical mutagenesis yield at present.ConclusionOur results showed that K+ and Ca2+ ions primarily limited the efficient production of ethanol. Then, subsequent comparative transcriptomic GO and pathway analyses showed that the co-presence of K+ and Ca2+ exerted the most prominent limitation on efficient ethanol production. The results of this study might prove useful by promoting the development and utilization of green fuel bio-manufactured from molasses.

Comprehensive Evaluation of Fuel Ethanol Production from Poplar Wood Fermentation and its Coupled Pathway with Lignin Gasification Based on Energy-Environment-Economy

Comprehensive Evaluation of Fuel Ethanol Production from Poplar Wood Fermentation and its Coupled Pathway with Lignin Gasification Based on Energy-Environment-Economy