High-gravity fermentation, used for ethanol production from sugarcane, corn, and mixed substrates, offers several benefits. Yeast, a rapidly multiplying unicellular microorganism, can be adapted for high sugar and ethanol tolerance on a lab scale. However, different substrates can enhance fermentation efficiency. Our study consisted of two experiments. In the first, we compared simple batch feeding with a fed-batch system for yeast selection in high-gravity fermentation. We ran eight cycles with increasing initial sugar contents (50 to 300 g L⁻¹). No significant differences were observed in the first seven cycles, but in the eighth, the fed-batch system showed lower glycerol and fructose contents and higher cell viability than the simple batch system. In the second experiment, we used the fed-batch system with 300 g L⁻¹ from sugarcane, corn, and mixed wort. The results showed that mixed wort produced higher ethanol contents and greater fermentation efficiency compared to corn and sugarcane as substrates. In conclusion, our findings indicate that the fed-batch system is more suitable for high-gravity fermentation on a lab scale, and the combination of sugarcane juice and corn can enhance fermentation efficiency, paving the way for integrating these substrates in industrial ethanol production.

This study investigated improvements in sugarcane ethanol production by adapting yeast strains for very high gravity fermentation. Two yeast strains (C22 and Y904) were adapted in eight fermentation cycles with increasing initial sugar content from 56.2 to 296.1 g L⁻¹ (Experiment 1). After the last cycle, the “adapted” yeasts were recycled in a wort containing 296.1 g L⁻¹ initial sugar and compared with their respective strains that were not subjected to the adaptation process (Experiment 2). Fermentative parameters were analyzed and the osmotic stress on yeast cell morphology was assessed by scanning electron microscopy (SEM). In Experiment 1, along the fermentation cycles, strain Y904 showed a decrease in cell viability after sugar concentration of 223 g L⁻¹. SEM images showed that Y904 cells were wrinkled after this cycle. In the case of strain C22, no differences in cell viability were observed along the cycles. However, for both strains, the residual sugars were relatively high and the ethanol content was below the maximum potential. In Experiment 2, for strain Y904, no differences were observed between adapted and non-adapted yeasts in terms of ethanol content, cell viability, and morphology. In the case of strain C22, cell viability and final ethanol content were significantly higher in the adapted yeast, which had cells less damaged by the osmotic stress. In conclusion, the study supports the importance of yeast strain selection and adaptation for efficient VHG fermentation, by demonstrating the superior performance of yeast strain C22 in response to increasing initial sugar content.

Despite being considered renewable, corn (Zea mays) ethanol still generates much debate over the use of fossil fuels in its production and is considered less sustainable than sugarcane (Saccharum spp.) ethanol. In Brazil, corn ethanol is starting to be produced in the Center-West and is expected to increase with the RenovaBio, a promising policy for biofuels adoption. In this context, energy cane (Saccharum spp.) is a biomass crop with high yields that can provide bagasse to supply the energy demand of the corn ethanol industry and provide juice with about 10% sugar content. However, the effects of introducing its juice in the production process are unknown. For these reasons, the objective of this study was to assess the effects of adding energy cane juice in corn ethanol production. Energy cane juice brings several advantages: (i) It provides sugars that can reduce by almost 50% the amount of corn and enzymes used, (ii) reduces the amount of water needed for ethanol production, and (iii) increases significantly the fermentation efficiency from 86.4% to 90.8% by providing minerals that support yeast growth. Therefore, energy cane can be integrated into the corn ethanol production process, making the fermentation more efficient and the production systems more sustainable.

Recently, in Brazil, corn ethanol industries are being installed and the integration with sugar/energy-cane has been proposed, using bagasse for cogeneration and the juice to dilute the corn. However, this integration may have some limitations, such as the quality of the cane juice and potential contamination by microorganisms brought with the cane from the field. In this article, we first tested the effects of mixing energy cane juice with corn on fermentative parameters. We also assessed the effects of Lactobacilli. contamination on organic acids produced during the fermentation and fermentation parameters and proposed the use of ionizing radiation to replace antibiotics as a disinfection control method. Our results showed that mixing energy cane juice with corn does not have any negative effect on fermentation parameters, including ethanol production. The contamination with Lactobacilli. considerably increased the production of acetic, lactic, and succinic acid, reducing the pH and ethanol content from 89.2 g L⁻¹ in the sterilized treatment to 72.9 g L⁻¹ in the contaminated treatment. Therefore, for the integration between corn and cane to be applied on an industrial scale, it is essential to have effective disinfection before fermentation. Ionizing radiation (20 kGy) virtually disinfected the wort, showing itself to be a promising technology; however, an economic viability study for adopting it in the industry should be carried out.

Yeast recycling, which is a common practice in sugarcane ethanol plants, could be expanded if it could be successfully implemented in corn-based ethanol production. However, the process of recycling the yeast remaining after fermentation is hampered by contaminating microorganisms that reduce the fermentation efficiency and compete with the yeast for the fermentable sugars. Currently, antibiotics are used to control microbial contamination. This study proposes chlorine dioxide and electron beam irradiation as alternative control methods for completely inactivating contaminants and minimizing their effect on recycled yeast. For that, wort sterilization using electron radiation (20 kGy) and treatment with a chemical biocide, namely chlorine dioxide (125 mg kg⁻¹), were compared with non-treated wort. Five fermentation cycles were performed using fed-batch systems with 300 g L⁻¹ of fermentable sugars. The results obtained in this study showed the inactivation of contaminants under the effect of electron beam irradiation, which led to an increase in the productivity, yield, and efficiency of fermentation by 0.21 g L⁻¹h⁻¹, 2.6%, and 4.7%, respectively. However, ClO₂ did not show promising results in reducing contamination or improving fermentative parameters. Thus, electron beam irradiation of contaminated wort may be a suitable alternative to chemical biocides and would allow the use of recycled yeast in corn-based ethanol plants.