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Abstract An imminent change in the world energy matrix makes it necessary to increase the production of renewable fuels. The United States and Brazil are the world's largest producers, but their production methods are very different, using different raw materials, ground corn and sugarcane juice, respectively. In recent years, strong investments have been made to expand the use of corn in Brazilian ethanol production. The combination of the sugar cane and corn ethanol industries has generated innovations in the sector, such as the "flex" mills, which are traditional sugar cane mills adapted to produce corn ethanol in the sugar cane off-season. Brazil has a portfolio of robust industrial yeasts for sugarcane ethanol production, naturally evolved and selected over the past 50 years. In this work, we analyze for the first time the performance of Brazilian industrial strains (BG-1, CAT-1, PE-2 and SA-1, widely used in sugarcane ethanol production) in corn ethanol production using different stress conditions. Ethanol Red yeast, traditionally used in corn ethanol plants around the world, was used as a control. In terms of tolerance to temperature (35 °C), strains BG-1 and SA-1 stood out. In fermentations with high solids concentration (35%), strain BG-1 reached ethanol contents higher than 19% w/v and had a productivity gain of 5.8% compared to fermentation at 30%. This was the first time that these industrial strains were evaluated using the high solids concentration of 35% and the results point to ways to improve the corn ethanol production process. Graphical Abstract

Dark fermentation (DF) is a promising biological process for hydrogen production from biomass. However, low hydrogen yield (HY) is a major hurdle impeding its use at large-scale operation. A potential way to mitigate this problem is to increase the concentration of substrate in the process to increase hydrogen production. The present study investigated the possibility of using high-solid DF to produce hydrogen from cassava processing wastes, i.e. , cassava pulp (CP) and cassava processing wastewater (CPW). CP was suspended in CPW and hydrolyzed enzymatically under optimum conditions of 150 g-CP/L, 29 U/g of α-amylase, 47 U/g of glucoamylase, and 60 FPU/g of cellulase. The hydrolysis performed at 50 °C for 24 h yielded a reducing sugar concentration of 117.7 ± 1.8 g/L, equivalent to 0.78 g-reducing-sugar/g-CP. Subsequent DF of CP-CPW enzymatic slurry, which contained 12.1% water insoluble solids, resulted in a cumulative production of 13.72 ± 0.22 L-H 2 , equivalent to 225.2 ± 3.7 mL-H 2 /g-VS. This was 83.1% of a maximum stoichiometric HY, based on carbohydrate content of CP and soluble metabolites production. The present study shows clearly the applicability of high-solid DF in the production of hydrogen from cassava processing wastes. • Hydrogen production from cassava pulp (CP) and cassava processing wastewater (CPW). • Enzymatic hydrolysis of CP-CPW suspension was statistically optimized. • Proper mixing improved hydrogen production in high-solid dark fermentation. • Hydrogen yield (HY) equivalent to 83.1% of a stoichiometric maximum HY was attained. [ABSTRACT FROM AUTHOR]

An imminent change in the world energy matrix makes it necessary to increase the production of renewable fuels. The United States and Brazil are the world's largest producers, but their production methods are very different, using different raw materials, ground corn and sugarcane juice, respectively. In recent years, strong investments have been made to expand the use of corn in Brazilian ethanol production. The combination of the sugar cane and corn ethanol industries has generated innovations in the sector, such as the "flex" mills, which are traditional sugar cane mills adapted to produce corn ethanol in the sugar cane off-season. Brazil has a portfolio of robust industrial yeasts for sugarcane ethanol production, naturally evolved and selected over the past 50 years. In this work, we analyze for the first time the performance of Brazilian industrial strains (BG-1, CAT-1, PE-2 and SA-1, widely used in sugarcane ethanol production) in corn ethanol production using different stress conditions. Ethanol Red yeast, traditionally used in corn ethanol plants around the world, was used as a control. In terms of tolerance to temperature (35 °C), strains BG-1 and SA-1 stood out. In fermentations with high solids concentration (35%), strain BG-1 reached ethanol contents higher than 19% w/v and had a productivity gain of 5.8% compared to fermentation at 30%. This was the first time that these industrial strains were evaluated using the high solids concentration of 35% and the results point to ways to improve the corn ethanol production process. [ABSTRACT FROM AUTHOR]

Synergistic effect of cellulase and hemicellulase (xylanase) was evaluated because lignocellulosic material is a heterogeneous complex of cellulose and hemicellulose. Various effects of HTec2 addition on enzymatic saccharification and fermentation were evaluated using two different substrates such as corn stover and rice straw. Corn stover and rice straw were pretreated by the LMAA (low-moisture anhydrous ammonia) method at the preselected same conditions (90 °C, 120 h, moisture content = 50%, NH3 loading = 0.1 g NH3/g). It was observed that the enzymatic saccharification yield of pretreated corn stover (76.4% for glucan digestibility) was higher than that of pretreated rice straw (70.9% for glucan) using CTec2 cellulase without HTec2 addition. Glucan digestibility of pretreated corn stover was significantly increased from 76.4% to 91.1% when the HTec2/CTec2 (v/v) increased from 0 to 10. However, it was interesting that the ethanol production was decreased from 89.9% to 76.3% for SSF and 118.0% to 87.9% for SSCF at higher HTec2/CTec2. As the glucan loading increased from 2.0% to 7.0%, the ethanol yields of both SSF and SSCF were decreased from 96.3% to 88.9% and from 116.6% to 92.4%, respectively. In addition, the smallest inoculum size (optical density of 0.25) resulted in the highest ethanol production (20.5 g/L).

Steam explosion coupling high-temperature short-time sterilization (SE-HTST) was exploited to modify cellulosic biomass medium properties and promote high-solid fermentation (HSF). Biomass characterization analysis showed that SE-HTST enlarged microstructural pores and cavities in solid media, providing more effective space for microbial growth. Meanwhile, SE-HTST helped to release glucose from the cellulose with 35.8 ± 4.5, 20.0 ± 2.3, and 12.3 ± 5.7 mg glucose/g dry medium at 24, 48, and 72 h of fermentation, which were 3.1, 2.3, and 1.5 times higher than that in medium from conventional thermal sterilization (CTS), respectively. SE-HTST increased the viable cell and spore number of Bacillus subtilis by 1.8 and 1.6 times at 72 h of fermentation compared to CTS. Moreover, the expressions of master transcriptional gene spo0A and the early sigma factors of sigF and sigE genes gradually increased in the SE-HTST medium, showing enhanced sporulation in HSF. Therefore, SE-HTST is an effective strategy for facilitating cellulose degradation, improving glucose nutrients in biomass medium, and promoting sporulation-regulatory gene expression during high-solid fermentation, which enhances the production of microbial ecological agents using B. subtilis significantly. • SE-HTST facilitated cellulose degradation and improved nutrients in biomass media. • High levels of spo0A, sigF, and sigE expression with SE-HTST indicated enhanced sporulation. • SE-HTST presented a porous medium and effective contact space for microbial cells. • The proposed model well elucidated microstructural changes of cellulosic biomass media. • SE-HTST improved biomass medium properties and facilitated fermentation performance. [ABSTRACT FROM AUTHOR]

Background: Conventional corn dry-grind ethanol production process requires exogenous alpha and glucoamylases enzymes to breakdown starch into glucose, which is fermented to ethanol by yeast. This study evaluates the potential use of new genetically engineered corn and yeast, which can eliminate or minimize the use of these external enzymes, improve the economics and process efficiencies, and simplify the process. An approach of in situ ethanol removal during fermentation was also investigated for its potential to improve the efficiency of high-solid fermentation, which can significantly reduce the downstream ethanol and co-product recovery cost. Results: The fermentation of amylase corn (producing endogenous α-amylase) using conventional yeast and no addition of exogenous α-amylase resulted in ethanol concentration of 4.1 % higher compared to control treatment (conventional corn using exogenous α-amylase). Conventional corn processed with exogenous α-amylase and superior yeast (producing glucoamylase or GA) with no exogenous glucoamylase addition resulted in ethanol concentration similar to control treatment (conventional yeast with exogenous glucoamylase addition). Combination of amylase corn and superior yeast required only 25 % of recommended glucoamylase dose to complete fermentation and achieve ethanol concentration and yield similar to control treatment (conventional corn with exogenous α-amylase, conventional yeast with exogenous glucoamylase). Use of superior yeast with 50 % GA addition resulted in similar increases in yield for conventional or amylase corn of approximately 7 % compared to that of control treatment. Combination of amylase corn, superior yeast, and in situ ethanol removal resulted in a process that allowed complete fermentation of 40 % slurry solids with only 50 % of exogenous GA enzyme requirements and 64.6 % higher ethanol yield compared to that of conventional process. Conclusions: Use of amylase corn and superior yeast in the dry-grind processing industry can reduce the total external enzyme usage by more than 80 %, and combining their use with in situ removal of ethanol during fermentation allows efficient high-solid fermentation. [ABSTRACT FROM AUTHOR]

Synergistic effect of cellulase and hemicellulase (xylanase) was evaluated because lignocellulosic material is a heterogeneous complex of cellulose and hemicellulose. Various effects of HTec2 addition on enzymatic saccharification and fermentation were evaluated using two different substrates such as corn stover and rice straw. Corn stover and rice straw were pretreated by the LMAA (low-moisture anhydrous ammonia) method at the preselected same conditions (90 °C, 120 h, moisture content = 50%, NH3 loading = 0.1 g NH3/g). It was observed that the enzymatic saccharification yield of pretreated corn stover (76.4% for glucan digestibility) was higher than that of pretreated rice straw (70.9% for glucan) using CTec2 cellulase without HTec2 addition. Glucan digestibility of pretreated corn stover was significantly increased from 76.4% to 91.1% when the HTec2/CTec2 (v/v) increased from 0 to 10. However, it was interesting that the ethanol production was decreased from 89.9% to 76.3% for SSF and 118.0% to 87.9% for SSCF at higher HTec2/CTec2. As the glucan loading increased from 2.0% to 7.0%, the ethanol yields of both SSF and SSCF were decreased from 96.3% to 88.9% and from 116.6% to 92.4%, respectively. In addition, the smallest inoculum size (optical density of 0.25) resulted in the highest ethanol production (20.5 g/L). [ABSTRACT FROM AUTHOR]

A lignin amphoteric surfactant and betaine could enhance the enzymatic hydrolysis of lignocellulose and recover cellulase. The effects of lignosulfonate quaternary ammonium salt (SLQA) and dodecyl dimethyl betaine (BS12) on enzymatic hydrolysis digestibility, ethanol yield, yeast cell viability, and other properties of high-solid enzymatic hydrolysis and fermentation of a corncob residue were studied in this research. The results suggested that SLQA and 1 g/L BS12 effectively improved the ethanol yield through enhancing enzymatic hydrolysis. SLQA had no significant effect on the yeast cell membrane and glucose fermentation. However, 5 g/L BS12 reduced the ethanol yield as a result of the fact that 5 g/L BS12 damaged the yeast cell membrane and inhibited the conversion of glucose to ethanol. Our research also suggested that 1 g/L BS12 enhanced the ethanol yield of corncob residue fermentation, which was attributed to the fact that lignin in the corncob adsorbed BS12 and decreased its concentration in solution to a safe level for the yeast.