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The study aims to enhance ethanol production by Wickerhamomyces subpelliculosus ZE75 isolated from marine sediment. In addition, analyzing the kinetic parameters of ethanol production and optimization of the fermentation conditions was performed. The marine yeast isolate ZE75 was selected as the front runner ethanol-producer, with an ethanol yield of 89.77 gL−1. ZE75 was identified relying on the phenotypic and genotypic characteristics of W. subpelliculosus. The genotypic characterization based on the Internal Transcribed Spacer (ITS) sequence was deposited in the GenBank database with the accession number OP715873. The maximum specific ethanol production rate (vmax) was 0.482 gg−1 h−1 at 175 gL−1 glucose concentration, with a high accuracy of R2 0.95. The maximum growth specific rates (μmax) were 0.141 h−1 obtained at 150 gL−1 glucose concentration with R2 0.91. Optimization of the fermentation parameters such as pH and salinity has been achieved. The highest ethanol yield 0.5637 gg−1 was achieved in a 100% natural seawater-based medium. The maximum ethanol production of 104.04 gL−1 was achieved at pH 4.5 with a specific ethanol rate of 0.1669 gg−1 h−1. The findings of the present study recommend the possibility of ethanol production from a seawater-based medium on a large scale using W. subpelliculosus ZE75. [ABSTRACT FROM AUTHOR]

In this article, the problem of real‐time estimation of the fermentative ethanol process is tackled. The considered observer is a model‐based technique that is robust regarding the model parameter uncertainties and inline noisy measurements. An unstructured kinetic model was used to describe the production of ethanol in a batch bioreactor for Saccharomyces cerevisiae. The biomass concentration was selected as the measured bioreactor´s output via an inline device, where the estimate variables were the substrate and ethanol concentrations. An experimental prototype was constructed to demonstrate the observer's real‐time performance. The experimental results show that the robust, smooth sliding mode observer performs better than the standard proportional sliding mode observer. [ABSTRACT FROM AUTHOR]

• SCF as a kind of periodic operation is analyzed for its superiority over the steady-state counterpart. • SCF as a kind of impulsive control is evaluated for possible repetition property in long-term operation. • Optimal on-off operation strategies are formulated with guaranteed periodic stability and superiority. Fuel ethanol is highlighted for being renewable and carbon neutral, but a multi-scale challenge exists that hampers its role as a future energy substitute, and with the breakthrough of cellulose pretreatment and enzyme hydrolysis technology, fermentation itself has become one of the main limiting factors for process enhancement. To tackle the problem, this paper studies periodic operation strategies to improve process performance and avoid possible instabilities. Firstly, superiority by periodic operation over the corresponding steady-state operation is verified. Then, to settle the problem of possible instabilities under impulsive control, stability of state-impulsive control is conducted and for the studied case, any self-cycling fermentation starts from batch mode with 50 % discharge-and-refill portion exhibits superior repeatability property, which allows continuous fermentation to operate under very slow overall dilution rate to cope with the extreme slow fermentation rate. Finally, on-off operation problem is formulated that is governed by a multidimensional nonlinear system, affine to the manipulated variable that is under the isoperimetric constraint; with the use of Pontryagin maximum principle, the optimal synthesis of the optimal strategies under integral constraint is computed out, and a practically plausible on-off operation strategy for ethanol fermentation is formulated with guaranteed periodic stability and optimality. [ABSTRACT FROM AUTHOR]

• Thermally self-sufficient heat pump-assisted dividing-wall column for biofuel recovery. • Eco-efficient product recovery after isopropanol – butanol – ethanol (IBE) fermentation. • Cost-effective (0.126 $/kgIBE) and energy-efficient (0.673 kWeh/kgIBE) downstream processing. • Heat pumping and heat integration for electrification of industrial biotechnology. • Large-scale downstream process for sustainable industrial biofuels production. Isopropanol-butanol-ethanol (IBE) fermentation is a superior biofuel production technology as compared to acetone-butanol-ethanol (ABE) fermentation due to the better fuel properties of the obtained products. However, low product concentrations, thermodynamic constraints and the presence of microorganisms lead to complex downstream processing that limits the competitiveness of this biofuel production method. Thus, this original research proposes a novel thermally self-sufficient and eco-efficient downstream process for industrial-scale recovery after IBE fermentation (74 ktonne/y capacity), from a highly dilute broth (>97 wt% water). Gas stripping and heat pump-assisted vacuum evaporation were implemented to separate valuable products from most of the broth. Furthermore, an advanced highly integrated heat pump-assisted azeotropic dividing-wall column was designed to recover high-purity (99 wt%) butanol biofuel and isopropanol – ethanol fuel supplement (89 wt%). The proposed purification process recovers over 99 % of biofuel products in a cost-effective (0.130 $/kg IBE) and energy-efficient way (0.673 kW e h/kg IBE) while allowing full recycle of biomass and most of the separated water. Besides improving yield by continuously recovering the inhibitory products, fermentation can be further enhanced by avoiding biomass loss and reducing water requirements. Lastly, the implemented energy-saving techniques ensure complete electrification of the proposed IBE recovery process. Therefore, the original results of this research study significantly contribute to the development of sustainable biofuel production processes. [Display omitted] [ABSTRACT FROM AUTHOR]

Waterlogging stress is one of the major abiotic stresses affecting the productivity and quality of many crops worldwide. However, the mechanisms of waterlogging tolerance are still elusive in barley. In this study, we identify key differentially expressed genes (DEGs) and differential metabolites (DM) that mediate distinct waterlogging tolerance strategies in leaf and root of two barley varieties with contrasting waterlogging tolerance under different waterlogging treatments. Transcriptome profiling revealed that the response of roots was more distinct than that of leaves in both varieties, in which the number of downregulated genes in roots was 7.41-fold higher than that in leaves of waterlogging sensitive variety after 72 h of waterlogging stress. We also found the number of waterlogging stress-induced upregulated DEGs in the waterlogging tolerant variety was higher than that of the waterlogging sensitive variety in both leaves and roots in 1 h and 72 h treatment. This suggested the waterlogging tolerant variety may respond more quickly to waterlogging stress. Meanwhile, phenylpropanoid biosynthesis pathway was identified to play critical roles in waterlogging tolerant variety by improving cell wall biogenesis and peroxidase activity through DEGs such as Peroxidase (PERs) and Cinnamoyl-CoA reductases (CCRs) to improve resistance to waterlogging. Based on metabolomic and transcriptomic analysis, we found the waterlogging tolerant variety can better alleviate the energy deficiency via higher sugar content, reduced lactate accumulation, and improved ethanol fermentation activity compared to the waterlogging sensitive variety. In summary, our results provide waterlogging tolerance strategies in barley to guide the development of elite genetic resources towards waterlogging-tolerant crop varieties. [ABSTRACT FROM AUTHOR]

Background: The use of ionic liquids (ILs) to fractionate lignocelluloses for various bio-based chemicals productions is in the ascendant. On this basis, the protic ILs consisting of triethylammonium hydrogen sulfate ([TEA][HSO4]) possessed great promise due to the low price, low pollution, and high efficiency. In this study, the microwave-assistant [TEA][HSO4] fractionation process was established for corn stover fractionation, so as to facilitate the monomeric sugars production and supported the downstream acetone–butanol–ethanol (ABE) fermentation. Results: The assistance of microwave irradiation could obviously shorten the fractionation period of corn stover. Under the optimized condition (190 W for 3 min), high xylan removal (93.17 ± 0.63%) and delignification rate (72.90 ± 0.81%) were realized. The mechanisms for the promotion effect of the microwave to the protic ILs fractionation process were ascribed to the synergistic effect of the IL and microwaves to the depolymerization of lignocellulose through the ionic conduction, which can be clarified by the characterization of the pulps and the isolated lignin specimens. Downstream valorization of the fractionated pulps into ABE productions was also investigated. The [TEA][HSO4] free corn stover hydrolysate was capable of producing 12.58 g L−1 of ABE from overall 38.20 g L−1 of monomeric sugars without detoxification and additional nutrients supplementation. Conclusions: The assistance of microwave irradiation could significantly promote the corn stover fractionation by [TEA][HSO4]. Mass balance indicated that 8.1 g of ABE and 16.61 g of technical lignin can be generated from 100 g of raw corn stover based on the novel fractionation strategy. [ABSTRACT FROM AUTHOR]

Despite the significant progress in the research, the problem of finding an efficient method for producing bioethanol from renewable lignocellulosic waste materials remains unresolved. Our investigation aimed to assess the efficacy of ethanol production from sugar beet pulp (SBP) utilising various approaches, including pretreatment variations, enzymatic processes, and microbial hydrolysis. Our research involved using the post-cultivation concentrate of T. viride LOCK 0588 grown in the SBP-based medium as a source of enzymes. The SBP hydrolysis process was carried out for 48 h at 50 °C. The quantity of sugar released, up to 61 g dm−3, through the utilisation of this extract proved to be on par with the outcomes achieved by the application of the commercial Cellic Ctec2 preparation. The final yields of the ethanol production with the use of the coculture of S. cerevisiae (Ethanol Red) and Scheffersomyces stipitis LOCK 0047 strain were in the range 5.1 ± 0.11 kg 100 kg−1 ÷ 5.38 ± 0.11 kg 100 kg−1. These results provide a solid basis for improving larger-scale industrial procedures that involve converting SBP into bioethanol using a cost-efficient approach of microbial hydrolysis with T. viride and a blend of pentose and hexose fermenting yeast. [ABSTRACT FROM AUTHOR]

Growing demand for renewable and sustainable energy sources has prompted research into bioethanol production as a viable alternative to hydrocarbon fuel for automobile engines. Based on their high carbohydrate content, Chlorella vulgaris microalgae were chosen for this study as a feasible biomass for bioethanol synthesis. After conducting microwave-based acid hydrolysis with varying acid concentrations of sulfuric acid (H2SO4) for 1–15 min, this study found that 3% H2SO4 for 5 min yielded the highest yield of reducing sugar (6.77 g/L). In this work, the RSM approach of central composite design (CCD) was utilized for modeling and optimization of ethanol fermentation and the MATLAB Simulink approach was employed for the simulation of ethanol fermentation. This study compared the effects of using Saccharomyces cerevisiae yeast for fermentation at different fermentation times (12 to 48 h), temperatures (28 to 32°C), and size of inoculum (0.5–1.5 g/L). Based on the actual and predicted results, the best conditions for fermentation were 36 h of time, 30°C of temperature, and 1.5 g/L of inoculum size. This gave the maximum ethanol concentration of 3.31 g/L. Furthermore, the RSM method provides minimum error and maximum R² value results for ethanol production compared to MATLAB simulation prediction (MSP). In addition, residue biomass from biofuel production is evaluated for its suitability for a further form of biofuel production. This research explores the opportunities for zero-waste biomass utilization in the development of biofuels. [ABSTRACT FROM AUTHOR]

[Display omitted] • A combined fractionation with surfactants and diluted EDA was developed. • PEG8000 addition significantly enhanced lignin removal (68.9%) in the solid fraction. • PEG8000 promoted the preservation of β-O-4 bonds and aliphatic hydroxyl groups in the fractionated lignin. • 137.2 g of ABE solvents could be obtained from 1 kg dried corn stover. In this study, a surfactant-assisted diluted ethylenediamine (EDA) fractionation process was investigated for co-generation of technical lignin and biobutanol from corn stover. The results showed that the addition of PEG 8000 significantly enhanced cellulose recovery (88.9 %) and lignin removal (68.9 %) in the solid fraction. Moreover, the pulp achieved 86.5 % glucose yield and 82.6 % xylose yield in enzymatic hydrolysis. Structural characterization confirmed that the fractionation process promoted the preservation of active β-O-4 bonds (35.8/100R) in isolated lignin and functionalized the lignin through structural modification using EDA and surfactant grafting. The enzymatic hydrolysate of the pulps yielded a sugar solution for acetone-butanol-ethanol (ABE) fermentation, resulting in an ABE concentration of 15.4 g/L and an overall yield of 137.2 g/Kg of dried corn stalk. Thus, the surfactant-assisted diluted EDA fractionation has the potential to enhance the overall economic feasibility of second-generation biofuels production within the framework of biorefinery. [ABSTRACT FROM AUTHOR]

Background: Saccharomyces 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. Results: In 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. Conclusion: Our 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. [ABSTRACT FROM AUTHOR]

The scarcity of essential minerals in the wort used for bioethanol production has long been a persistent concern in industrial processes. This scarcity can negatively impact various aspects of production, including yeast cell vitality and sugar-to-ethanol conversion. In this study, we assessed the nutritional profile of key components of the industrial bioethanol production process and identified nitrogen, phosphorus, magnesium, manganese, zinc, and copper ions deficiencies. Supplementation of fermentation assays with TMNitrofós was carried out until reaching the maximum allowed for manganese (24.5%), zinc (49.4%), and copper (8.8%). Laboratory-scale fermentation assays were conducted under varying conditions of soluble solids in the wort (20, 24, and 26°Brix). These experiments aimed to assess the primary productivity parameters in response to the application of TMNitrofós. Results showed that fermentations using wort with 24 and 26°Brix supplemented with TMNitrofós led to 7% higher fermentative efficiency compared to the control group. The supplemented fermentations demonstrated higher levels of cellular vitality (76.6%) compared to the control group (55.3%). Considering a scale-up in the bioethanol production process with TMNitrofós supplementation, an increase of 266 m3 of ethanol per month was achieved compared to the control group, resulting in cost savings of R$ 586,879.20.Graphical Abstract: The scarcity of essential minerals in the wort used for bioethanol production has long been a persistent concern in industrial processes. This scarcity can negatively impact various aspects of production, including yeast cell vitality and sugar-to-ethanol conversion. In this study, we assessed the nutritional profile of key components of the industrial bioethanol production process and identified nitrogen, phosphorus, magnesium, manganese, zinc, and copper ions deficiencies. Supplementation of fermentation assays with TMNitrofós was carried out until reaching the maximum allowed for manganese (24.5%), zinc (49.4%), and copper (8.8%). Laboratory-scale fermentation assays were conducted under varying conditions of soluble solids in the wort (20, 24, and 26°Brix). These experiments aimed to assess the primary productivity parameters in response to the application of TMNitrofós. Results showed that fermentations using wort with 24 and 26°Brix supplemented with TMNitrofós led to 7% higher fermentative efficiency compared to the control group. The supplemented fermentations demonstrated higher levels of cellular vitality (76.6%) compared to the control group (55.3%). Considering a scale-up in the bioethanol production process with TMNitrofós supplementation, an increase of 266 m3 of ethanol per month was achieved compared to the control group, resulting in cost savings of R$ 586,879.20. [ABSTRACT FROM AUTHOR]

Agricultural residues are more suitable for biochemical conversion to produce bulk commodities preferentially through anaerobic fermentation with less energy consumption and lower contamination risk. Steam explosion is applicable for industrial production, and biological pretreatment is not suitable for fermentation under pure culture conditions due to its slow rate and contamination risk. Cellulases produced in situ and formulated with glucosidases are cost-effective for building up sugar platforms through simultaneous saccharification and co-fermentation (SSCF) and separate hydrolysis and co-fermentation (SHCF) to produce volatile and nonvolatile products, respectively. In situ utilization of CO 2 can credit cellulosic ethanol, and both yield and titer must be improved substantially for butanol to be produced as an advanced biofuel, but microbial lipids produced aerobically with intensive energy consumption and low yield cannot be affordable as feedstock for biodiesel production. Lignocellulose is an alternative to fossil resources, but its biochemical conversion is not economically competitive. While decentralized processing can reduce logistical cost for this feedstock, sugar platforms need to be developed with energy-saving pretreatment technologies and cost-effective cellulases, and products must be selected correctly. Anaerobic fermentation with less energy consumption and lower contamination risk is preferred, particularly for producing biofuels. Great effort has been devoted to producing cellulosic ethanol, but CO 2 released with large quantities during ethanol fermentation must be utilized in situ for credit. Unless titer and yield are improved substantially, butanol cannot be produced as an advanced biofuel. Microbial lipids produced through aerobic fermentation with low yield and intensive energy consumption are not affordable as feedstocks for biodiesel production. [ABSTRACT FROM AUTHOR]

Summary: This study focuses primarily on the real‐time monitoring of reducing sugar levels throughout solid‐state ethanol fermentation. A predictive model for accurate quantification of reducing sugar levels will be formulated through a comprehensive analysis of near‐infrared spectra across different fermentation stages. Building a predictive model using Adaptive Boosting (AdaBoost), The spectral data was pre‐processed using the Standard Normal Variate (SNV) method, followed by feature band selection using the Genetic Algorithm (GA), resulting in the identification of 181 characteristic wavelengths. The resulting GA‐AdaBoost model has excellent performance metrics: Rc2 of 0.989, RMSEC of 0.393, Rp2 of 0.978, RMSEP of 0.640, RPD of 6.651 and RER of 0.142. Comparative evaluations against Competitive Adaptive Reweighted Sampling (CARS), Successive Projection Algorithm (SPA), Uninformative Variable Elimination (UVE) and Principal Component Analysis (PCA) consistently underscore the superior efficacy of the GA‐AdaBoost model. These results robustly confirm its reliability in accurately quantifying reducing sugars in solid‐state fermentation processes. [ABSTRACT FROM AUTHOR]

Saccharomyces cerevisiae is a widely used strain for ethanol fermentation; meanwhile, efficient utilization of glucose could effectively promote ethanol production. The PFK1 gene is a key gene for intracellular glucose metabolism in S. cerevisiae. Our previous work suggested that although deletion of the PFK1 gene could confer higher oxidative tolerance to S. cerevisiae cells, the PFK1Δ strain was prone to contamination by other microorganisms. High interspecies microbial competition ability is vital for the growth and survival of microorganisms in co-cultures. The result of our previous studies hinted us a reasonable logic that the EMP (i.e., the Embden-Meyerhof-Parnas pathway, the glycolytic pathway) key gene PFK1 could be involved in regulating interspecies competitiveness of S. cerevisiae through the regulation of glucose utilization and ethanol production efficiency. The results suggest that under 2% and 5% glucose, the PFK1Δ strain showed slower growth than the S288c wild-type and TDH1Δ strains in the lag and exponential growth stages, but realized higher growth in the stationary stage. However, relative high supplement of glucose (10%) eliminated this phenomenon, suggesting the importance of glucose in the regulation of PFK1 in yeast cell growth. Furthermore, during the lag growth phase, the PFK1Δ strain displayed a decelerated glucose consumption rate (P < 0.05). The expression levels of the HXT2, HXT5, and HXT6 genes decreased by approximately 0.5-fold (P < 0.05) and the expression level of the ZWF1 exhibited a onefold increase in the PFK1Δ strain compared to that in the S. cerevisiae S288c wild-type strain (P < 0.05).These findings suggested that the PFK1 inhibited the uptake and utilization of intracellular glucose by yeast cells, resulting in a higher amount of residual glucose in the medium for the PFK1Δ strain to utilize for growth during the reverse overshoot stage in the stationary phase. The results presented here also indicated the potential of ethanol as a defensive weapon against S. cerevisiae. The lower ethanol yield in the early stage of the PFK1Δ strain (P < 0.001) and the decreased expression levels of the PDC5 and PDC6 (P < 0.05), which led to slower growth, resulted in the strain being less competitive than the wild-type strain when co-cultured with Escherichia coli. The lower interspecies competitiveness of the PFK1Δ strain further promoted the growth of co-cultured E. coli, which in turn activated the ethanol production efficiency of the PFK1Δ strain to antagonize it from E. coli at the stationary stage. The results presented clarified the regulation of the PFK1 gene on the growth and interspecies microbial competition behavior of S. cerevisiae and would help us to understand the microbial interactions between S. cerevisiae and other microorganisms. Key points: • PFK1Δ strain could realize reverse growth overshoot at the stationary stage • PFK1 deletion decreased ethanol yield and interspecific competitiveness • Proportion of E. coli in co-culture affected ethanol yield capacity of yeast cells [ABSTRACT FROM AUTHOR]

Background: The abundance of glucuronoxylan (GX) in agricultural and forestry residual side streams positions it as a promising feedstock for microbial conversion into valuable compounds. By engineering strains of the widely employed cell factory Saccharomyces cerevisiae with the ability to directly hydrolyze and ferment GX polymers, we can avoid the need for harsh chemical pretreatments and costly enzymatic hydrolysis steps prior to fermentation. However, for an economically viable bioproduction process, the engineered strains must efficiently express and secrete enzymes that act in synergy to hydrolyze the targeted polymers. Results: The aim of this study was to equip the xylose-fermenting S. cerevisiae strain CEN.PK XXX with xylanolytic enzymes targeting beechwood GX. Using a targeted enzyme approach, we matched hydrolytic enzyme activities to the chemical features of the GX substrate and determined that besides endo-1,4-β-xylanase and β-xylosidase activities, α-methyl-glucuronidase activity was of great importance for GX hydrolysis and yeast growth. We also created a library of strains expressing different combinations of enzymes, and screened for yeast strains that could express and secrete the enzymes and metabolize the GX hydrolysis products efficiently. While strains engineered with BmXyn11A xylanase and XylA β-xylosidase could grow relatively well in beechwood GX, strains further engineered with Agu115 α-methyl-glucuronidase did not display an additional growth benefit, likely due to inefficient expression and secretion of this enzyme. Co-cultures of strains expressing complementary enzymes as well as external enzyme supplementation boosted yeast growth and ethanol fermentation of GX, and ethanol titers reached a maximum of 1.33 g L− 1 after 48 h under oxygen limited condition in bioreactor fermentations. Conclusion: This work underscored the importance of identifying an optimal enzyme combination for successful engineering of S. cerevisiae strains that can hydrolyze and assimilate GX. The enzymes must exhibit high and balanced activities, be compatible with the yeast's expression and secretion system, and the nature of the hydrolysis products must be such that they can be taken up and metabolized by the yeast. The engineered strains, particularly when co-cultivated, display robust growth and fermentation of GX, and represent a significant step forward towards a sustainable and cost-effective bioprocessing of GX-rich biomass. They also provide valuable insights for future strain and process development targets. [ABSTRACT FROM AUTHOR]

The co-cultivation of sake yeast (AK25, K901, K1401, or K1801 strain) and the kuratsuki Bacillus A-10 and/or Priestia B-12 strains in koji solution was performed to demonstrate the effects of these two kuratsuki bacteria on sake taste. The results showed that the Brix and acidity patterns of sake preparations produced with and without these kuratsuki bacteria were very similar. This indicated that the addition of these kuratsuki bacteria did not inhibit ethanol fermentation or organic acid production by sake yeast. A taste recognition device showed that the effects of these kuratsuki bacteria on the saltiness and sourness of sake were greater than those on other taste properties. Astringency stimulation and saltiness of sake produced using the sake yeast K901 were increased by Bacillus A-10 and decreased by Priestia B-12. Except for these two cases, the taste intensities of sake preparations produced with the Bacillus A-10 and Priestia B-12 strains were very similar, but differed from those of sake produced with kuratsuki Kocuria. These results support our hypothesis that the flavor and taste of sake can be controlled by utilizing the interactions between kuratsuki bacteria and sake yeast. For crating the desired sake taste, a combination of kuratsuki bacteria and sake yeast should be considered. [ABSTRACT FROM AUTHOR]

Bioethanol is an organic compound resulted from the fermentation of sugar substrates by microorganisms which is used as alternative energy sources. During bioethanol fermentation yeast are exposed to various fermentation stresses, including temperature, osmotic, and oxidative stresess. Such conditions may decrease ethanol production. We previously isolated fermentation-stress tolerance yeast isolates from traditional Balinese beverages, identified as Wickerhamomyces anomalus BT2, BT5, and BT6. However no data available regarding the bioethanol production of those isolates. Our study indicates that these strains could utilize various sugar substrates (glucose, xylose, maltose, sucrose) in oxidative fermentative media. The highest value of substrate utilization efficiency following 48 hours fermentation was shown by BT6 on glucose (61.02%), BT 2 on xylose (55.44%) and maltose (60.90%). Measurement of ethanol production by Gas Chromatography showed that the strains were able to produce higher ethanol on the glucose substrate than other substrates. For instance, BT6 could produce the highest ethanol production (5.00 g/L) amongst strains tested by using glucose as substrate. Yet, the particular strains could only produce 0.30 g/L and 0.65 g/L by using xylose and maltose, respectively. For further genetic engineering purposes, we detected ADH1 gene from all three isolates, with high homology to the alcohol dehydrogenase from Saccharomyces cerevisiae, Geobacillus stearothermophilus and Pseudomonas aeruginosa. Further strain development can be carried out targeting the ADH1 gene, important for ethanol fermentation. [ABSTRACT FROM AUTHOR]

To improve ethanol production, carbon and microbial sources were derived from corncob and molasses. Before investigating simultaneous saccharification and fermentation (SSF), the boiled corncob was vacuum-ensiled and the molasses was diluted. The optimal ensiling time of 4 days increased not only the amount of fermentable sugar but also the population of beneficial microorganisms. The proper dilution of molasses at a ratio of 1:3 molasses to boiled water reduced both the contamination problem and the inhibition of ethanol fermentation. The combination of the ensiled corncob and the diluted molasses could provide a substrate that is suitable for SSF without the addition of enzymes, supplements, or yeast strains. The substrate contained the optimal concentration of initial sugar (174.631 ± 0.975 g/L) and viable microorganisms (1.9×106 ± 0.1×106 CFU/mL of total bacteria and 1.0x105 ± 0.1×105CFU/mL of yeast) that could continue to hydrolyze and ferment carbohydrates in the substrate for the SSF. Using a ratio of 1:3 of ensiled corncob to diluted molasses, a pH of 4.5, and a temperature of 27 °C for 72 h, the SSF produced the highest concentration of ethanol at 93.345 ± 0.062 g/L. This work provides a cost-effective process that requires no enzyme or yeast, is simply technology, and is friendly to the environment, while the remaining solid fraction can be utilized as a suitable feedstock for biogas production. [ABSTRACT FROM AUTHOR]

Saccharomyces cerevisiae is a widely used strain for ethanol fermentation; meanwhile, efficient utilization of glucose could effectively promote ethanol production. The PFK1 gene is a key gene for intracellular glucose metabolism in S. cerevisiae. Our previous work suggested that although deletion of the PFK1 gene could confer higher oxidative tolerance to S. cerevisiae cells, the PFK1Δ strain was prone to contamination by other microorganisms. High interspecies microbial competition ability is vital for the growth and survival of microorganisms in co-cultures. The result of our previous studies hinted us a reasonable logic that the EMP (i.e., the Embden-Meyerhof-Parnas pathway, the glycolytic pathway) key gene PFK1 could be involved in regulating interspecies competitiveness of S. cerevisiae through the regulation of glucose utilization and ethanol production efficiency. The results suggest that under 2% and 5% glucose, the PFK1Δ strain showed slower growth than the S288c wild-type and TDH1Δ strains in the lag and exponential growth stages, but realized higher growth in the stationary stage. However, relative high supplement of glucose (10%) eliminated this phenomenon, suggesting the importance of glucose in the regulation of PFK1 in yeast cell growth. Furthermore, during the lag growth phase, the PFK1Δ strain displayed a decelerated glucose consumption rate (P < 0.05). The expression levels of the HXT2, HXT5, and HXT6 genes decreased by approximately 0.5-fold (P < 0.05) and the expression level of the ZWF1 exhibited a onefold increase in the PFK1Δ strain compared to that in the S. cerevisiae S288c wild-type strain (P < 0.05).These findings suggested that the PFK1 inhibited the uptake and utilization of intracellular glucose by yeast cells, resulting in a higher amount of residual glucose in the medium for the PFK1Δ strain to utilize for growth during the reverse overshoot stage in the stationary phase. The results presented here also indicated the potential of ethanol as a defensive weapon against S. cerevisiae. The lower ethanol yield in the early stage of the PFK1Δ strain (P < 0.001) and the decreased expression levels of the PDC5 and PDC6 (P < 0.05), which led to slower growth, resulted in the strain being less competitive than the wild-type strain when co-cultured with Escherichia coli. The lower interspecies competitiveness of the PFK1Δ strain further promoted the growth of co-cultured E. coli, which in turn activated the ethanol production efficiency of the PFK1Δ strain to antagonize it from E. coli at the stationary stage. The results presented clarified the regulation of the PFK1 gene on the growth and interspecies microbial competition behavior of S. cerevisiae and would help us to understand the microbial interactions between S. cerevisiae and other microorganisms. Key points: • PFK1Δ strain could realize reverse growth overshoot at the stationary stage • PFK1 deletion decreased ethanol yield and interspecific competitiveness • Proportion of E. coli in co-culture affected ethanol yield capacity of yeast cells [ABSTRACT FROM AUTHOR]

[Display omitted] • Structures of DESs affected the pretreatment efficiency and ethanol fermentation. • Lactic acid/guanidine hydrochloride effectively removed lignin and xylan. • The hydrolysates of various DES pulps had impressive ethanol yields. • 302.75 g ethanol and 118.98 g lignin were harvested from 1 kg dried biomass. Deep eutectic solvents (DESs) offer a potential opportunity in biomass utilization industries. This work emphasized the impact of hydrogen bond donors (HBD) and acceptors (HBA) on deconstruction and valorization of rice straw. Acidity, alkyl chain length, hydrogen bonding ability and functional groups of HBD and HBA appeared to be important factors affecting the fractionated pulps and lignins, which further influenced ethanol fermentation. Among the candidate DESs, lactic acid/guanidine hydrochloride (LGH) was proved to be the most suitable one due to the excellent delignification and xylan removal. For the downstream fermentation process, 0.47 g g−1 of bioethanol with 0.55 g/L h−1 of productivity can be obtained from the LGH pulp's hydrolysate. Mass balance showed 302.8 g bioethanol and 119.0 g technical lignin can be co-generated from 1 kg dried rice straw. This "green" valorization strategy offers a promising scheme in biorefinery of lignocelluloses. [ABSTRACT FROM AUTHOR]

In this study, sorghum meal underwent 2–10 kGy electron beam irradiation (EBI) to investigate alterations in its elemental composition, relevant physicochemical characteristics, and enzymatic hydrolysis fermentation properties. The results indicated a decrease in amylose content and an increase in reducing sugar content in sorghum meal after EBI, along with severe damage to the internal structure of starch granules, leading to increased solubility in water. Furthermore, irradiation enhanced the shear resistance and gelatinization stability of sorghum meal paste. Most importantly, EBI-induced degradation of sorghum starch molecules enhanced their susceptibility to interact with α-amylase, significantly improving the enzymatic hydrolysis and alcohol fermentation efficiency of sorghum meal. At an irradiation dose of 8 kGy, raw and cooked sorghum meal exhibited a 43.40% and 33.33% increase in ethanol concentration and a 43.44% and 30.29% increase in ethanol yield, respectively. This finding indicated that irradiation is an effective pretreatment method for starch-based ethanol fermentation. [Display omitted] • Using EBI as a pretreatment for sorghum meal ethanol fermentation. • Sorghum starch molecules degraded after EBI treatment. • EBI notably enhanced sorghum meal's enzymatic saccharification efficiency. • The ethanol concentration and yield peaked at an EBI dose of 8 kGy. • EBI coupled with high-temperature cooking notably improved fermentation efficiency. [ABSTRACT FROM AUTHOR]

The WGS of Klebsiella sp. SWET4 revealed a genome of 5,665,821 bp by assembling 78 scaffolds with N50 and L50 values of 371571 and 5, respectively. BUSCO and ortholog analysis indicated the completeness of the genome and familiarity with Klebsiella variicola. Annotation result confirmed the presence of 5 major cellulose metabolizing genes (bcsZ, bglC, bglA, celA, chbA), besides 4 key xylan degrading genes (xynB, xynT, xylA, xylB) and 4 principal ethanol fermentation (nifJ, adhE, acs, adh1) genes. The lignolytic potential of SWET4 was evident in the kinetic study and the presence of yfeX / efeB , katG , katE , etc. genes were confirmed. Expression study with qPCR indicated overexpression of cellulolytic genes (268.73-fold of bglA , 122.78-fold of bglC , and 45.88-fold of bcsZ) in cellulose broth when compared to glucose. However, less activity was observed for xylanolytic genes (9.19-fold of xylA , 4.29-fold of xylB , and 2.89-fold of xynB) in xylan broth. Additionally, the kinetic study revealed that K s was significantly lower for cellulose (7.79 and 0.09 g/L) compared to xylan (193.78 and 3.95 g/L) for primary and secondary growth phases, respectively which confirmed the higher preference of cellulose by SWET4. Further, ethanol estimation by HPLC exhibited an increase in ethanol concentration in banana peel fermentation broth from 0.1 g/L (30 h) to 0.35 g/L (56 h). The adh1 gene expression was also found to increase by 84.45-fold, whereas adhE increased by 1.62-fold confirming their influence on ethanol production. This study confirmed the lignocellulolytic and ethanologenic properties of SWET4 enabling the direct biotransformation of cellulosic biomass to ethanol. [Display omitted] • The WGS of Klebsiella sp. SWET4 was reviled the genome size of 5665821 bp. • Completeness of the constructed genome was studied by BUSCO and ortholog analysis. • bglA exhibited 268.73-fold and xylA 9.19-fold of overexpression by qPCR study. • adh1 exhibited 84.4-fold increase resulting 35-fold increase in ethanol production. • Metabolic pathway from plant polysaccharide to ethanol was established. [ABSTRACT FROM AUTHOR]

Pretreatment of lignocellulosic biomass by breaking down its complex structure is an essential step to facilitate enzymatic hydrolysis of lignocellulose and subsequent bioethanol fermentation. Herein, an eco-friendly pretreatment strategy was developed by applying deep eutectic solvent (DES) cocktail. Attributing to the synergistic effect of two DESs (e.g., betaine-glycerol and choline chloride-oxalic acid (or choline chloride-acetamide)), an impressive 89.51% digestibility for glucan and 75.43% for xylan were achieved. Characterization of pretreated corn stover showed that DES cocktail changed substrate surface structure and enlarged structure pores, which could improve the accessibility of cellulase. The pretreated corn stover was also employed in ethanol fermentation and achieved the yield of 73.35%. Recycling experiments demonstrated the potential reusability of DES cocktail in a sustainable manner towards various biomass sources. These findings highlight the effectiveness and versatility of the developed DES cocktail pretreatment method, paving the way for sustainable bioethanol production from lignocellulosic feedstocks. [Display omitted] • Betaine-glycerol based DES cocktail was developed for corn stover pretreatment. • DES cocktail pretreatment increased glucan recovery up to 13.1% compared to single DES pretreatment. • The saccharification rate of enzyme-digested glucose after optimization of pretreatment temperature was as high as 89.51%. • Ethanol yield reached 73.61% with a maximum increase of 10.16% after DES cocktail pretreatment. • DES cocktails maintained ∼83% pretreatment effect after four recovery cycles. [ABSTRACT FROM AUTHOR]

Several lignocellulosic wastes are generated in the pulp and paper industry (PPI), such as small wood chips (pinchips) and paper sludge, presenting a high cellulose content suitable to be converted into biofuels or bio-products in a forest biorefinery scheme. In this work, two schemes of biorefinery were proposed for their valorization, processing small eucalyptus wood pinchips in two different strategies: (i) autohydrolysis at 230ºC, and (ii) autohydrolysis at 195ºC followed by organosolv process (47.7% ethanol-water, 198ºC for 60 min). More than 95% of cellulose was recovered in both schemes. In the combined process, 76% of delignification was achieved and 78% of xylan was solubilized as xylooligosaccharides. To reduce operational cost of lignocellulosic biomass-to-ethanol fermentation, the mixture of the treated eucalyptus pinchips from two processes with sludge was also proposed to increase the initial glucan content and to supply a rich source of nitrogen (present in the sludge). For that, two experimental designs were carried out for ethanol production by simultaneous saccharification and fermentation (SSF) process. Ethanol from SSF assays using sludge as co-substrate at 0.6 g of sludge/g of treated wood pinchips and 16 FPU/g of pretreated solids allowed to obtain 59 g/L (90% of conversion) and 46 g/L (96% of conversion) when blended with the wood from autohydrolysis and with the wood from autohydrolysis followed by organosolv, respectively. Overall, this study shows an alternative process valorization of biomasses derived from PPI for production of advanced biofuels and bio-products (such as xylooligosaccharides and lignin) contributing to achieving a circular economy. • Multi-waste valorization approach was proposed to produce biofuel and bioproducts. • Autohydrolysis and organosolv processes allowed > 90% glucan recoveries. • Higher than 40 g/L of ethanol and 90% of yield were obtained in both schemes developed. • Blending with sludge improve ethanol production from pretreated eucalyptus wood. [ABSTRACT FROM AUTHOR]

Biodegradable mulch film has become an encouraging alternative to conventional petroleum–based plastic, but it is likely to generate more microplastics (MPs) than conventional films with lower resistance and thus profoundly affects the soil environment. However, the effects of biodegradable (BMPs) and conventional polyethylene MPs (PE–MPs) on soil microbial carbon (C) cycle remain unclear. Here, metagenomic sequencing was used to investigate microbial functional and relative taxonomic traits in response to PE–MP and BMP addition (5 % w/w) via a soil microcosm experiment. The results showed that the presence of BMPs, rather than PE–MPs, significantly changed soil microbial C cycling patterns at the functional and taxonomic levels. In soils, BMP addition significantly increased the abundances of genes regulating aerobic respiration, C decomposition (intracellular), C fixation, fermentation and CO oxidation pathways compared to those in the control and PE–MP added soils. Specifically, BMPs promoted soil starch and PHA (liable C) degradation, acetate and ethanol fermentation, Calvin–Benson–Bassham (CBB) cycle and 3–hydroxypropionate/4–hydroxybutyrate (3HP/4HB) associated with C fixation. Most of these C cycling pathways were further strengthened in the microplastisphere of BMPs. Furthermore, MPs selectively enriched specific taxa on their surface and subsequently increased their abundance in soils; Bradyrhizobium , Ramlibacter and Variovorax (enriched by BMPs) positively regulated labile C degradation, and Amycolatopsis and Nocardia (enriched by PE–MPs) promoted recalcitrant C degradation. Moreover, Pseudonocardia and Variovorax that were enriched by BMPs improved C dissimilation and assimilation by regulating fermentation and C fixation processes, respectively. Taken together, these results improve the understanding of the influence of conventional and biodegradable MPs on soil C cycling pathways and related microbial communities, highlighting that the presence of BMPs rather than PE–MPs accelerates soil C turnover. • BMPs rather than PE–MPs significantly affected microbial functions involved in soil C cycle. • BMPs addition promoted multiple processes of microbial carbon metabolisms. • Proteobacteria and its genera enriched by BMPs improved liable C degradation. • PE – MPs enriched Amycolatopsis and Nocardia that regulated recalcitrant C decomposition. • Pseudonocardia and Variovorax significantly regulated fermentation and C fixation metabolisms. [ABSTRACT FROM AUTHOR]

Sugarcane molasses is one of the main raw materials for bioethanol production, and Saccharomyces cerevisiae is the major biofuel-producing organism. In this study, a batch fermentation model has been used to examine ethanol titers of deletion mutants for all yeast nonessential genes in this yeast genome. A total of 42 genes are identified to be involved in ethanol production during fermentation of sugarcane molasses. Deletion mutants of seventeen genes show increased ethanol titers, while deletion mutants for twenty-five genes exhibit reduced ethanol titers. Two MAP kinases Hog1 and Kss1 controlling the high osmolarity and glycerol (HOG) signaling and the filamentous growth, respectively, are negatively involved in the regulation of ethanol production. In addition, twelve genes involved in amino acid metabolism are crucial for ethanol production during fermentation. Our findings provide novel targets and strategies for genetically engineering industrial yeast strains to improve ethanol titer during fermentation of sugarcane molasses. • Screening 4797 gene mutants for ethanol fermentation titers of sugarcane molasses. • Forty-two genes involved in ethanol production in sugarcane molasses fermentation. • Seventeen mutants show increased, but 25 mutants exhibit reduced, ethanol yields. • Two MAP kinases Hog1 and Kss1 negatively regulate ethanol production. • Amino acid metabolism are crucial for ethanol production during fermentation. [ABSTRACT FROM AUTHOR]

The unwashed alkali-treated lignocellulose can be directly enzymatically hydrolyzed and fermented via pH adjustment with acids. The use of acids would give a burden on production cost. Furfural residue (FR) which is the acidic solid waste from lignocellulose-derived furfural production process was employed in this study as a pH regulator. The corn cob-derived FR was used to adjust the pH value of alkali-treated corn stover (PCS) to 4.8 for enzymatic hydrolysis and ethanol fermentation. The unwashed PCS adjusted by FR got higher enzymatic hydrolysis efficiency (EHE) than the washed PCS samples. Meanwhile, the mixing of PCS and FR had a synergistic effect on the EHE of PCS. The fermentation of enzymatic hydrolysate from unwashed PCS-FR mixture at 20% solid concentration could attain ethanol production of 26.54 ± 0.02 mg/mL with a yield of 89.53 ± 0.08%. This work created a novel recycling way of FR as a pH regulator for improving the bioconversion of alkali-treated lignocellulose. It also provided a novel clue for the valuable valorization of wastes from corn production. [ABSTRACT FROM AUTHOR]

• Upgrading DAP by post-hydrolysis for carbon efficient conversion of lignocellulose. • A radar chart method was used for the comprehensive evaluation of DAP performance. • One-pot post-hydrolysis and SSF without detoxification and sterilization. • Preserved 45.2 % of original β-O-4 linkages in fractionated lignin. • 66 % of lignocellulose carbon was recovered as ethanol, xylose and reactive lignin. Dry acid pretreatment (DAP) as a promising process for industrial biorefinery provide an efficient bioconversion of cellulose without free wastewater, although the partial xylan and lignin degrade to inhibitors or recondense. A biorefinery strategy for carbon efficient conversion of lignocellulose into bioethanol, xylose, and reactive lignin was developed by upgrading DAP with post-hydrolysis. The results showed that lignocellulose after mild DAP (175 °C, acid dosage of 15 mg/g dry material) obtained higher xylan recovery and lower inhibitors than that of general DAP. Subsequently, post-hydrolysis, simultaneous saccharification and ethanol fermentation were performed at solids loading of 20 wt% without detoxification and sterilization, resulting in xylose and ethanol yield of 71.8 % and 67.6 %. The fractionated lignin presented more reactive β-aryl ether linkages and less condensation than that from DAP. 66 % of lignocellulose carbon was recovered as ethanol, xylose and reactive lignin. This upgrading biorefinery strategy provided an easy-to-operate process for integrated utilization of lignocellulose. [ABSTRACT FROM AUTHOR]

Lipid-extracted algae (LEA) can be processed via acetone, butanol and ethanol fermentation to produce butanol as an additive to algal biofuel, but direct fermentation was ineffective. In this study, nitrogen and phosphorus regulation for algae cultivation was applied to modify the biochemical composition of algal biomass and hence improve butanol production from LEA. The results show that when nitrogen starvation and excess phosphorus were used during algae cultivation, a significant rise of starch productivity and decline of protein productivity for algal biomass were obtained, which resulted in the highest butanol productivity of 20.16 mg L−1 d−1, a 5.48-fold increase over traditional one-stage cultivation. Mechanism analysis suggested that nitrogen starvation enhanced lipid and starch contents and reduced protein content, which modified C/N ratio of LEA, prevented acid crash and improved butanol production; excess phosphorus modified amino acid contents of phenylalanine and arginine, which strengthened fluxes directing to butanol generation. Through this investigation, the approach of using nitrogen starvation and excess phosphorus to modify biochemical composition of algal biomass and subsequently improve butanol production of LEA was demonstrated to be applicable. [Display omitted] [ABSTRACT FROM AUTHOR]

Hydrocotyle verticillata can tolerate varying degrees of flooding, up to complete submergence, and is at the same time extremely sensitive to drought. Understanding the structural and biochemical principles of these unusual tolerance limits is of particular importance. We analyzed the effect of soil flooding, complete submergence (rooted plants and floating stems), and dehydration on root anatomy, alcohol dehydrogenase (ADH), heat shock proteins 70 (HSP70), hydrogen peroxide, and DNA integrity using light microscopy, biochemical and histological methods. It was shown that anatomical traits of adventitious roots with a triarch stele were similar in the plants growing under different conditions; the single-layered and thin-walled epidermis formed relatively short root hairs; essential air spaces were absent in the cortex parenchyma. Results on ADH clearly showed that anaerobic energetic metabolism in root apices and individual rhizoderma cells of the mature root zone was normal for this species in optimal and suboptimal conditions, while leaves changed metabolism to anaerobic in response to submergence. Alterations in the protein spectrum were accompanied by adequate up-regulation of HSP70 under different levels of flooding and dehydration/rehydration. These results appear to indicate a flooding adaptation strategy for H. verticillata based primarily on metabolic plasticity rather than morphoanatomical adaptations. Most notably, the resistance of this species to long-term submergence has been associated with strong ADH induction in leaves, transient activation of 70 kDa isoform of HSP70 and induction of 66 kDa isoform, as well as a significant delay in the accumulation of hydrogen peroxide and DNA degradation. • Hydrocotyle verticillata tolerates varying degrees of flooding up to submergence. • Essential air spaces were absent in the root cortex parenchyma. • Ethanol fermentation was normal for roots and was induced in submerged leaves. • Up-regulation of HSP70 responded to both flooding and dehydration. • Delayed ROS production and DNA degradation were associated with leaf flood resistance. [ABSTRACT FROM AUTHOR]