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The pretreatment and saccharification of dewaxed bagasse (DWB) has been investigated under various reaction conditions ranging 2000 to 3200 psi, at 70 ± 1 °C in supercritical carbon dioxide (SCC). This has been in attempt to transform the DWB into fermentable sugar and bioethanol in high yields. The effect of SCC mediated pretreatment and enzymatic hydrolysis on structural and morphological alterations in DWB has been ascertained through diverse analytical methods. The sugar has been released through cellulase (40 FPU/mL) mediated enzymatic hydrolysis of pretreated DWB in sodium acetate buffer (pH 4.7) within 1 h at SCC 2800 psi, 70 ± 1 °C. The released sugar was subsequently fermented in the presence of yeast (Saccharomyces crevices, 135 CFU) at 28 ± 1 °C over 72 h to afford the bioethanol. The SCC mediated process conducted in acetic acid:water media (1:1) at 2800 psi, 70 ± 1 °C over 6 h has afforded the pretreated DWB with maximum yield towards the production of fermentable sugar and bioethanol. The production of fermentable sugar and bioethanol has been electrochemically estimated through cyclic voltammetry (CV) and square wave voltammetry (SWV) over glassy carbon electrode in KOH (0.1 M). The electrochemical methods were found selective and in close agreement for estimation of the yields (%) of fermentable sugars and bioethanol. The yield (%) of fermentable sugar estimated from CV and SWV were 80.10 ± 5.34 and 79.00 ± 5.09 respectively. Whereas the yield (%) of bioethanol estimated from CV and SWV were 81.30 ± 2.78% and 78.6 ± 1.25% respectively. Present investigation delivers a SCC mediated green and sustainable method of pretreatment of DWB to afford the enhanced saccharification, to produce bioethanol in high yields.

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Due to the electrical nature of the cell, it is possible to modulate its behavior through the application of non-lethal external electric fields to improve fermentation processes. In this work, a microbial cell system with a chamber and two electrodes inside and connected to a voltage source was used. One of the electrodes was kept isolated to create an electric field without the flow of current. Cultures with two ethanol-producing microbial strains (Saccharomyces cerevisiae and Zymomonas mobilis) were conducted in this device. The application of voltages between 0 and 18 V was evaluated to determine the impact of the generated electric field on ethanol production. To analyze the possible effect of the field on the central carbon metabolism in each strain, biochemical-based kinetic models were formulated to describe the experimental fermentation kinetics obtained. It was found that low applied voltages did not have significant effects on growth rate in either strain, but all voltages evaluated increased substrate consumption and ethanol production rate in Z. mobilis, while only 18 V affected these rates in S. cerevisiae, indicating that Z. mobilis was the most sensitive to the electric field. At the end of the fermentation, significant increases in ethanol yields of 10.7% and 19.5% were detected for S. cerevisiae and Z. mobilis, respectively. The proposed mathematical models showed that substrate transport through the membrane catalyzed by the phosphotransferase system (PTS) for Z. mobilis and hexose transport proteins mechanism and hexokinase (HK) activity for S. cerevisiae and the transformation of pyruvate to ethanol, catalyzed by the decarboxylase (PDC) and alcohol dehydrogenase (ADH) enzymes, were the reactions most affected by the application of the external field.

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Various thermochemical and biochemical processes are resorted to transform agri-wastes into diverse green fuels. Current investigation encompassed three different types of biomass viz., gingelly, kodo millet and horse grams, whose desirability as biofuel feedstock have been largely unexplored till date. The existence of significant amount of cellulose (38.07 %), volatiles (75.19 %), calorific value (avg. 16.98 MJ/kg) in the gingelly biomass, demonstrates the effectiveness of the concerned biomass for utilization as feedstock in diverse industrial applications. The mean estimates of Eα were lower for kodo millet (approx. 150 kJ/mole), followed by gingelly (approx. 178 kJ/mole) and horse gram (approx. 180 kJ/mole). The mean estimates of ΔHα were 174.81 (FWO), 170.22 (KAS), 169.17 (S) and 170.40 (T) kJ/mol for the gingelly biomass. The mean estimates of ΔHα were 147.83 (FWO), 148.81 (KAS), 147.93 (S) and 149.04 (T) kJ/mol for kodo millet biomass, while for horse gram biomass, mean estimates of ΔHα were 178.91 (FWO), 169.61 (KAS), 168.56 (S) and 168.81 (T) kJ/mol. The minor difference of 3-4 kJ/mole between Aα and Hα, signifies the viability of the thermal disintegration process. From master plot, it's evident that the experimental curve intersects multiple theoretical curves, highlighting the intricate characteristics of the thermal disintegration process. The overall ethanol recovery was highest in gingelly as compared to both the biomasses. Gingelly biomass yielded an ethanol titer of 24.8 g/L after 24 h, resulting in a volumetric ethanol productivity of 1.03 g/L/h and an ethanol yield of 0.36 g/g.

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Fossil resources must be replaced by renewable resources in production systems to mitigate green-house gas emissions and combat climate change. Electro-fermentation utilizes a bioelectrochemical system (BES) to valorize industrial and municipal waste. Current electro-fermentation research is mainly focused on microbial electrosynthesis using CO2 for producing commodity chemicals and replacing petroleum-based infrastructures. However, slow production rates and low titers of metabolites during CO2-based microbial electrosynthesis impede its implementation to the real application in the near future. On the other hand, CO is a highly reactive gas and an abundant feedstock discharged from fossil fuel-based industry. Here, we investigated CO and CO2 electro-fermentation, using a CO-enriched culture. Fresh cow fecal waste was enriched under an atmosphere of 50% CO and 20% CO2 in N2 using serial cultivation. The CO-enriched culture was dominated by Clostridium autoethanogenum (≥89%) and showed electro-activity in a BES reactor with CO2 sparging. When 50% CO was included in the 20% CO2 gas with 10 mA applied current, acetate and ethanol were produced up to 12.9 ± 2.7 mM and 2.7 ± 1.1 mM, respectively. The coulombic efficiency was estimated to 148% ± 8% without an electron mediator. At 25 mA, the culture showed faster initial growth and acetate production but no ethanol production, and only at 86% ± 4% coulombic efficiency. The maximum optical density (OD) of 10 mA and 25 mA reactors were 0.29 ± 0.07 and 0.41 ± 0.03, respectively, whereas it was 0.77 ± 0.19 without electric current. These results show that CO electro-fermentation at low current can be an alternative way of valorizing industrial waste gas using a bioelectrochemical system.

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Lignocellulosic biomass, the most abundant natural resource on earth, can be used for cellulosic ethanol production but requires a pretreatment to improve enzyme access to the polymeric sugars while obtaining value from the other components. γ-Valerolactone (GVL) is a promising candidate for biomass pretreatment since it is renewable and bio-based. In the present work, the effect of a pretreatment based on GVL on the enzymatic saccharification of white birch was evaluated at a laboratory scale and the importance of the washing procedure for the subsequent saccharification was demonstrated. Both the saccharification yield and the production of cellulosic ethanol were higher using a noncommercial enzyme crude from Talaromyces amestolkiae than with the commercial cocktail Cellic CTec2 from Novozymes. Furthermore, the production of extracellular cellulases by T. amestolkiae has been optimized in 2 L bioreactors, with improvements ranging from 40% to 75%. Finally, it was corroborated by isoelectric focus that optimization of cellulase secretion by T. amestolkiae did not affect the pattern production of the main ß-glucosidases and endoglucanases secreted by this fungus.

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The persistent expansion in world energy and synthetic compounds requires the improvement of renewable alternatives in contrast to non-sustainable energy wellsprings. Lignocellulose is an encouraging feedstock to be utilized in biorefineries for its conversion into value-added products, including biomaterials, biofuels and several bio-based synthetic compounds. Aside from all categories, biofuel, particularly bioethanol is the most substantial fuel derived from lignocellulosic biomass and can be obtained through microbial fermentation. Generally, extreme settings are required for lignocellulosic pretreatment which results in the formation of inhibitors during biomassdegradation. Occasionally, lignin polymers also act as inhibitors and are left untreated during the pretreatment, engendering inefficient hydrolysis. The valorization of lignocellulosic biomass by laccases can be viewed as a fundamental trend for improving bioethanol production. However, one of the main obstacles for developing commercially viable biofuel industries is the cost of enzymes, which can be resolved by utilizing laccases derived from microbial sources. Microbial laccases have been considered an exceptionally integral asset for delignification and detoxification of pretreated LCB, which amplify the resultant fermentation and saccharification processes. This review provides a summary of microbial laccases and their role in valorizing LCB to bioethanol, compelling enthralling applications in bio-refining industries all across the globe.

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In the context of biorefinery, researchers have been looking for lignocellulosic biomasses and ideal treatments to produce economically viable biofuels. In this scenario, the bamboo culm appears as a plant matrix of great potential, given the high cellulose content of low crystallinity. Thus, the objective and differential of this work was to determine the best conditions for enzymatic hydrolysis of cellulose extracted from bamboo culm and to evaluate its potential application in the production of bioethanol through Separate Hydrolysis and Fermentation (SHF) and Saccharification and Simultaneous Fermentation (SSF) by Saccharomyces cerevisiae modified via CRISPR/Cas9. The average cellulose extraction yield was 41.87 % with an extraction efficiency of 86.76 %. In general, as the hydrolysis time increased, an increase in glucose production was observed in almost all assays, with higher hydrolysis efficiency values at 72 h. The results ranged from 2.09 to 19.8 g/L of glucose obtained with efficiency values of 10.47 to 99 %. The best conditions were found in test 5 (temperature of 36 °C and pH 5.0, with only 10 FPU/g of substrate Cellic Ctec2 Novozymes ® cocktail). It is observed that for all hydrolysis times the independent variables pH and temperature were significant under the hydrolysis efficiency, showing a negative effect, indicating that higher values of the same promote lower values of the response variable. For bioethanol production, a maximum concentration of 7.84 g/L was observed for the SSH process after 4 h of fermentation, while for the SSF process it was 12.6 g/L after 24 h of fermentation, indicating the large potential of the simultaneous process together with the application of bamboo culm biomass for high production of biofuel.

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Sweet potato residue (SPR) is the by-product of starch extraction from fresh sweet potatoes and is rich in carbohydrates, making it a suitable substrate for bioethanol production. An amylolytic industrial yeast strain with co-expressing α-amylase and glucoamylase genes would combine enzyme production, SPR hydrolysis, and glucose fermentation into a one-step process. This consolidated bioprocessing (CBP) shows great application potential in the economic production of bioethanol. In this study, a convenient heterologous gene integration method was developed. Eight copies of a Talaromyces emersonii α-amylase expression cassette and eight copies of a Saccharomycopsis fibuligera glucoamylase expression cassette were integrated into the genome of industrial diploid Saccharomyces cerevisiae strain 1974. The resulting recombinant strains exhibited clear transparent zones in the iodine starch plates, and SDS-PAGE analysis indicated that α-amylase and glucoamylase were secreted into the culture medium. Enzymatic activity analysis demonstrated that the optimal temperature for α-amylase and glucoamylase was 60-70°C, and the pH optima for α-amylase and glucoamylase was 4.0 and 5.0, respectively. Initially, soluble corn starch with a concentration of 100 g/L was initially used to evaluate the ethanol production capability of recombinant amylolytic S. cerevisiae strains. After 7 days of CBP fermentation, the α-amylase-expressing strain 1974-temA and the glucoamylase-expressing strain 1974-GA produced 33.03 and 28.37 g/L ethanol, respectively. However, the 1974-GA-temA strain, which expressed α-amylase and glucoamylase, produced 42.22 g/L ethanol, corresponding to 70.59% of the theoretical yield. Subsequently, fermentation was conducted using the amylolytic strain 1974-GA-temA without the addition of exogenous α-amylase and glucoamylase, which resulted in the production of 32.15 g/L ethanol with an ethanol yield of 0.30 g/g. The addition of 20% glucoamylase (60 U/g SPR) increased ethanol concentration to 50.55 g/L, corresponding to a theoretical yield of 93.23%, which was comparable to the ethanol production observed with the addition of 100% α-amylase and glucoamylase. The recombinant amylolytic strains constructed in this study will facilitate the advancement of CBP fermentation of SPR for the production of bioethanol.

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In this study, the efficiency of a series of biochar-supported Cu catalysts, biochar-supported Zn catalysts, and biochar-supported Cu-Zn catalysts was determined through bioethanol dehydrogenation to the high-value chemical, acetaldehyde. Each metal, with weight percentages of 10, 20, and 30, and the combination of Cu-Zn, including 10 wt% of Cu and Zn, 15 wt% of Cu - 5 wt% of Zn, and 15 wt% of Cu and Zn, were fully loaded onto biochar using an incipient wetness impregnation technique. Subsequently, all biocatalysts were subjected to bioethanol dehydrogenation reactions in a temperature range of 200-400 °C. The optimum metal loading for the catalyst was found to be the combination of 15 wt% Cu and 15 wt% Zn. This catalyst resulted in a reasonable acetaldehyde yield of 56.2%, an initial bioethanol conversion of 57.3%, and a very high acetaldehyde selectivity of 98.1% at a mild reaction temperature of 300 °C and ambient pressure. These results were attributed to the optimal concentration of weak-medium acid and medium base sites. Active acid and base sites were identified through temperature-programmed desorption of ammonia (NH3-TPD) and temperature-programmed desorption of carbon dioxide (CO2-TPD), respectively. Furthermore, the reaction stability test of the best biocatalyst (15Cu-15Zn/BB) was proven by maintaining this reaction at the same temperature (300 °C) for 10 h. However, the catalytic performance slightly decreased due to the coke formation of Cu species.

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Crop straws provide enormous lignocellulose resources transformable for sustainable biofuels and valuable bioproducts. However, lignocellulose recalcitrance basically restricts essential biomass enzymatic saccharification at large scale. In this study, the mushroom-derived cellobiohydrolase (LeGH7) was introduced into Trichoderma reesei (Rut-C30) to generate two desirable strains, namely GH7-5 and GH7-6. Compared to the Rut-C30 strain, both engineered strains exhibited significantly enhanced enzymatic activities, with ß-glucosidases, endocellulases, cellobiohydrolases, and xylanase activities increasing by 113 %, 140 %, 241 %, and 196 %, respectively. By performing steam explosion and mild alkali pretreatments with mature straws of five bioenergy crops, diverse lignocellulose substrates were effectively digested by the crude enzymes secreted from the engineered strains, leading to the high-yield hexoses released for bioethanol production. Notably, the LeGH7 enzyme purified from engineered strain enabled to act as multiple cellulases and xylanase at higher activities, interpreting how synergistic enhancement of enzymatic saccharification was achieved for distinct lignocellulose substrates in major bioenergy crops. Therefore, this study has identified a novel enzyme that is active for simultaneous hydrolyses of cellulose and xylan, providing an applicable strategy for high biomass enzymatic saccharification and bioethanol conversion in bioenergy crops.

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This study focuses on investigating sugar recovery from spoiled date fruits (SDF) for sustainable ethanol production using newly isolated yeasts. Upon their isolation from different food products, yeast strains were identified through PCR amplification of the D1/D2 region and subsequent comparison with the GenBank database, confirming isolates KKU30, KKU32, and KKU33 as Saccharomyces cerevisiae; KKU21 as Zygosaccharomyces rouxii; and KKU35m as Meyerozyma guilliermondii. Optimization of sugar extraction from SDF pulp employed response surface methodology (RSM), varying solid loading (20-40%), temperature (20-40 °C), and extraction time (10-30 min). Linear models for sugar concentration (R1) and extraction efficiency (R2) showed relatively high R2 values, indicating a good model fit. Statistical analysis revealed significant effects of temperature and extraction time on extraction efficiency. The results of batch ethanol production from SDF extracts using mono-cultures indicated varying consumption rates of sugars, biomass production, and ethanol yields among strains. Notably, S. cerevisiae strains exhibited rapid sugar consumption and high ethanol productivity, outperforming Z. rouxii and M. guilliermondii, and they were selected for scaling up the process at fed-batch mode in a co-culture. Co-cultivation resulted in complete sugar consumption and higher ethanol yields compared to mono-cultures, whereas the ethanol titer reached 46.8 ± 0.2 g/L.

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Transitioning from a fossil-based to a bio-based economy is crucial to climate action and achieving neutrality in greenhouse gas (GHG) emissions. Biofuel production is an essential land-based GHG mitigation alternative. However, it raises concerns about biodiversity conservation, competition with food production, and net GHG emissions associated with direct land-use change (dLUC). This study aims to assess how the location and conversion routes influence GHG emissions for sugarcane expansion in Brazil to supply ethanol demand projections for 2030. A consistent and significant reduction in GHG emissions is achievable by implementing a strategy that prioritizes the spatial distribution for ethanol biorefinery expansions based on georeferenced life cycle emissions, including dLUC emissions associated with sugarcane production. Because of conservative zoning for sugarcane expansion, dLUC emissions are not an overriding factor, representing less than 9.1 % of the total GHG mitigation potential. Despite that, accounting for georeferenced dLUC emissions when prioritizing expansion facilities leads to spatial differences. Regarding conversion routes and land requirements, using cellulosic biorefineries could meet future projected demand based on sugarcane production from 3.1 million hectares, mostly in currently degraded pastureland. Conventional refineries would require 5.5 million hectares to meet the same demand of 71 billion liters. Despite the 77 % higher land demand to produce the same volume of ethanol, conventional refineries with straw recovery could be considered if electricity generation is a priority. This study illustrates how Brazil can achieve GHG mitigation targets while attending to future energy demand and protecting areas with high biodiversity.

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Bioethanol production from lignocellulosic materials is hindered by the high costs of pretreatment and the enzymes. The present study aimed to evaluate whether co-cultivation of four selected cellulolytic fungi yields higher cellulase and xylanase activities compared to the monocultures and to investigate whether the enzymes from the co-cultures yield higher saccharification on selected plant materials without thermo-chemical pretreatment. The fungal isolates, Trichoderma reesei F118, Penicillium javanicum FS7, Talaromyces sp. F113, and Talaromyces pinophilus FM9, were grown as monocultures and binary co-cultures under submerged conditions for 7 days. The cellulase and xylanase activities of the culture filtrates were measured, and the culture filtrates were employed for the saccharification of sugarcane leaves, Guinea grass leaves, and water hyacinth stems and leaves. Total reducing sugars and individual sugars released from each plant material were quantified. The co-culture of Talaromyces sp. F113 with Penicillium javanicum FS7 and of T. reesei F118 with T. pinophilus FM9 produced significantly higher cellulase activities compared to the corresponding monocultures whereas no effect was observed on xylanase activities. Overall, the highest amounts of total reducing sugars and individual sugars were obtained from Guinea grass leaves saccharified with the co-culture of T. reesei F118 with T. pinophilus FM9, yielding 63.5% saccharification. Guinea grass leaves were found to be the most susceptible to enzymatic saccharification without pre-treatment, while water hyacinth stems and leaves were the least. Accordingly, the study suggests that fungal co-cultivation could be a promising approach for the saccharification of lignocellulosic materials for bioethanol production.

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Enzymatic hydrolysis contributes to obtaining fermentable sugars using pretreated lignocellulose materials for bioethanol generation. Unfortunately, the pretreatment of lignocellulose causes low substrate enzymatic hydrolysis, which is due to the structure changes of lignin to produce main phenolic by-products and non-productive cellulase adsorption. It is reported that modified lignin enhances the speed of enzymatic hydrolysis through single means to decrease the negative effects of fermentation inhibitors or non-productive cellulase adsorption. However, a suitable modified lignin should be selected to simultaneously reduce the fermentation inhibitors concentration and non-productive cellulase adsorption for saving resources and maximizing the enzymatic hydrolysis productivity. Meanwhile, the adsorption micro-mechanisms of modified lignin with fermentation inhibitors and cellulase remain elusive. In this review, different pretreatment effects toward lignin structure, and their impacts on subsequent enzymatic hydrolysis are analyzed. The main modification methods for lignin are presented. Density functional theory is used to screen suitable modification methods for the simultaneous reduction of fermentation inhibitors and non-productive cellulase adsorption. Lignin-fermentation inhibitors and lignin-cellulase interaction mechanisms are discussed using different advanced analysis techniques. This article addresses the gap in previous reviews concerning the application of modified lignin in the enhancement of bioethanol production. For the first time, based on existing studies, this work posits the hypothesis of applying theoretical simulations to screen efficient modified lignin-based adsorbents, in order to achieve a dual optimization of the detoxification and saccharification processes. We aim to improve the integrated lignocellulose transformation procedure for the effective generation of cleaner bioethanol.

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Upgrading biomass-derived bioethanol to higher-order alcohols using conventional biotechnological approaches is challenging. Herein, a novel, magnetic metal-organic-framework-based cofactor regeneration system was developed using ethanol dehydrogenase (EtDH:D46G), NADH oxidase (NOX), formolase (FLS:L482S), and nicotinamide adenine dinucleotide (NAD+) for converting rice straw-derived bioethanol to acetoin. A magnetic zeolitic imidazolate framework-8@Fe3O4/NAD+ (ZIF-8@Fe3O4/NAD+) regeneration system for cell-free cascade reactions was introduced and used to encapsulate EtDH:D46G, NOX, and FLS:L482S (ENF). ZIF-8@Fe3O4/NAD+ENF created an efficient microenvironment for three-step enzyme cascades. Under the optimized conditions, the yield of acetoin from 100 mM bioethanol using ZIF-8@Fe3O4/NAD+ENF was 90.4 %. The regeneration system showed 97.1 % thermostability at 50 °C. The free enzymes retained only 16.3 % residual conversion, compared with 91.2 % for ZIF-8@Fe3O4/NAD+ENF after ten cycles. The magnetic metal-organic-framework-based cofactor regeneration system is suitable for enzymatic cascade biotransformations and can be extended to other cascade systems for potential biotechnological applications.

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The conversion of bioethanol to ethylene in gas phase and atmospheric pressure was investigated over γ-Al2O3 supported copper and nickel catalysts. These catalysts were prepared by co-precipitation and pre-treated with hydrogen at 450 °C. Six catalysts were studied at 450 °C under a nitrogen atmosphere. It was found that the monometallic Cu/γ-Al2O3 catalyst exhibited the highest ethylene concentration, with a selectivity of around 90 %. The bioethanol conversion obtained was between 57 %-86 %. Another catalyst that exhibited high concentration values was the NiCu1 : 7 bimetallic catalyst. The catalysts were characterised using XRD, SEM, EDS, TEM, TGA, FTIR, Raman, and N2-physisoption techniques. Furthermore, the Cu/γ-Al2O3 catalyst was studied under different reduction temperatures and gas flow conditions. It was found that the catalysts reduced at 350 °C and 35 ml/min N2 flow presented ethylene concentrations between (0.18-0.21) g/L. Moreover, the catalyst deactivation was identified to be first order and the equation of the Cu/γ-Al2O3 catalyst deactivation model was determined. Carbonaceous deposits over the used sample were not detected by Raman and FTIR. It was determined that the Cu/γ-Al2O3 catalyst deactivation could be mainly attributed to the blocking of the catalytic sites by strongly adsorbed compounds and hydroxylation of the catalyst surface.

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Microalgal biomass for biofuel production, integration into functional food, and feed supplementation has generated substantial interest worldwide due to its high growth rate, non-competitiveness for agronomic land, ease of cultivation in containments, and presence of several bioactive molecules. In this study, genetic engineering tools were employed to develop transgenic lines of freshwater microalga Chlorella vulgaris with a higher starch content, by up-regulating ADP-glucose pyrophosphorylase (AGPase), which is a rate-limiting enzyme in starch biosynthesis. Expression of the Escherichia coli glgC (AGPase homolog) gene in C. vulgaris led to an increase in total carbohydrate content up to 45.1% (dry cell weight, DCW) in the transgenic line as compared to 34.2% (DCW) in the untransformed control. The starch content improved up to 16% (DCW) in the transgenic alga compared to 10% (DCW) in the control. However, the content of total lipid, carotenoid, and chlorophyll decreased differentially in the transgenic lines. The carbohydrate-rich biomass from the transgenic algal line was used to produce bioethanol via yeast fermentation, which resulted in a higher ethanol yield of 82.82 mg/L as compared to 54.41 mg/L from the untransformed control. The in vitro digestibility of the transgenic algal starch revealed a resistant starch content of up to 7% of total starch. Faster growth of four probiotic bacterial species along with a lowering of the pH of the growth medium indicated transgenic alga to exert a positive prebiotic effect. Taken together, the study documents the utilization of genetically engineered C. vulgaris with enriched carbohydrates as bioethanol feedstock and functional food ingredients.

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The goal of the current work was to optimize the growth parameters needed to manufacture agarase enzyme from a non-marine PI strain of Bacillus subtilis on an agar-based medium. Using Plackett-Burman design (PBD), nine process parameters were evaluated, and agar, peptone, and yeast-extract were identified as the most significant independent factors influencing agarase production with confidence levels more than 90%. To evaluate the optimal concentrations of the indicated process parameters on agarase production, the Box-Behnken design (BBD) was applied. After optimization, B. subtilis strain PI produced 119.8 U/ml of agarase, representing a 1.36-fold increase. In addition the agar hydrolysate fermented products contain the liberated oligosaccharide acts as strong antioxidant which has 62.4% scavenging activity. Also, the agarase yields increased (1141.12, 1350.253, 1684.854 and 1921.863 U/ml) after substitution the agar with algal biomass of Carolina officinalis at different concentrations (2, 5, 10 and 15%), respectively. After completing the saccharification process, the resulted hydrolysate was used to produce ethanol through fermentation with Pichia pastoris yeast strain as an economical method giving yields (6.68317, 7.09748, 7.75648 and 8.22332 mg/ml), that are higher than using yeast extract peptone dextrose (YPD) medium (4.461 mg/ml).

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A substantial amount of process waste is generated during the manufacture of soft-wheat products (SWPs), such as biscuits/cookies, crackers, wafers, and cakes. A small portion of waste is reused in specific biscuits, whereas the rest is usually discarded. This study aimed to investigate the suitability of this waste for the co-production of bioethanol and fatty acid methyl esters (FAMEs or biodiesel). Two groups of waste generated in the SWP industry were included in the study: (a) the waste of low-moisture (<10%) biscuits, crackers, and wafer sheets with no fillings and/or coatings, and (b) the waste of high-moisture (>10%) biscuits, crackers, wafers, and cakes with fillings and/or coatings. The study involved extracting each sample with hexane, and the recovered fat was converted to the FAME through alkali-catalyzed transesterification. The remaining carbohydrate-rich fraction was then converted to bioethanol through amylolytic hydrolysis and yeast fermentation. A great portion (92.42%-93.17%) of the fat was extracted from the wastes and converted to the FAME with adequate yields (13.81-14.55 g FAME/g waste, dm) and acceptable conversion efficiencies (85.19%-89.04%). However, bioethanol production from the defatted carbohydrate-rich fractions proceeded rather slowly, yielding only 16.54-18.02 (g ethanol per g of waste, dm), corresponding to fermentation efficiencies ranging from 43.32% to 48.29%. Upon the co-production of FAME and ethanol, a considerable amount (50.93%-53.08%) of waste solids remained in the residue fraction. These findings indicated that production of the FAME with adequate yields and conversion efficiencies is viable from the SWP industry wastes; however, bioethanol yields and fermentation efficiencies are rather limited, which warrants further investigation. PRACTICAL APPLICATION: The soft-wheat processing industry generates 1%-5% of total production as waste. The waste was studied to produce FAME and bioethanol. The fat was extracted from the waste and converted to FAME. Bioethanol yields and fermentation efficiencies are limited due to dough modifiers and antimicrobial additives used in SWP production. Further research is required to improve ethanol yield.

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Seaweed, a valuable marine resource widely cultivated worldwide, can be vulnerable to stress and microbiome alterations, resulting in the decay of seaweeds and substantial economic losses. To investigate the seaweed-microbiome interaction, our study aimed to isolate marine bacteria and fungi that can cause Ice-Ice disease and evaluate their enzymatic characteristics for potential application in bioethanol production from seaweed biomass. Three red seaweed species (Gracilaria edulis, Kappaphycus alvarezii, and Eucheuma cottonii) were obtained for our study and placed in separate culture tanks. Among the 18 isolated marine microbial species, 12 tested positive for agar and carrageenan activity: six exhibited both activities, three displayed only agar activity, and three only carrageenan activity. DNA sequencing of the positive microbes identified ten bacteria and two yeast species. The 3,5-Dinitrosalicylic acid (DNSA) assay results revealed that the identified bacterial Caldibacillus kokeshiiformis strain FJAT-47861 exhibited the highest carrageenase activity (0.76 units/ml), while the yeast Pichia fermentans strain PM79 demonstrated the highest agarase activity (0.52 units/ml). Notably, Pichia fermentans strain PM79 exhibited the highest overall agarase and carrageenase activity, averaging 0.63 units/ml. The average carrageenase activity of all six positive microbes was 1.5 times higher than their agarase activity. These findings suggest that the 12 isolated microbes hold potential for bioethanol production from macroalgae, as their agarase and carrageenase activity indicates their ability to break down seaweed cell wall carbohydrates, causing ice-ice disease. Moreover, these results provide exciting prospects for harnessing the bioconversion capabilities of these microbes, paving the way for sustainable and efficient bioethanol production from seaweed resources. Supplementary Information: The online version contains supplementary material available at 10.1007/s12088-024-01205-w.