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The cellulase-producing Trichoderma reesei strain RL-P37 exhibits significant potential, yielding 7.3 g/L of cellulase in 241 hours. Microscopic investigations reveal a link between spore formation and enzyme production, suggesting the need for research into the intricate relationship between enzyme production, stress responses, and the nutritional prerequisites of fungi. Comparatively, the use of sodium hydroxide (NaOH) treatment, as opposed to water treatment, results in the reduction of micronutrient content and carbon source extraction as filtrate. Despite these challenges, research by He et al. (2021) highlights NaOH's efficiency in cellulose extraction from plant-based sources. Using NaOH pretreatment can be proven as effective by designing a proper cultivation method. The selection of inducers for enzyme induction gains importance, with soluble inducers, as emphasized by Zhang et al. (2022), exhibiting superior effectiveness. Hence, adopting soluble inducers in designing cultivation methods for improved enzyme production in shaking flasks is recommended. Enzymatic treatment of bio-waste, as outlined by Hu et al. (2021), shows promise in augmenting essential component content by breaking down plant cell walls and intercellular compartments. However, the feasibility of using an artificial bio-waste medium for cultivating Trichoderma reesei is questioned. Investigating the impact of micronutrient levels, particularly the inhibitory role of zinc, on fungal growth becomes essential. These findings underscore the necessity for ongoing research and optimization in cellulase production, emphasizing both strain productivity and cultivation methodologies.
As the population grows, so does the amount of biowaste. As demand for energy grows, biogas is a promising solution to the problem. Lignocellulosic materials are challenged of slow degradability due to the presence of polymers such as cellulose, lignin and hemicellulose. There are several pretreatment methods available to enhance the degradability of such materials, including enzymatic pretreatment. In this pretreatment, there are few parameters that can influence the results, the most important being the enzyme to solid ratio and the solid to liquid ratio. During this project, experiments were conducted to determine the optimal conditions for those two factors. It was discovered that a solid to liquid ratio of 31 g of buffer per 1 gram of organic dry matter produced the highest reducing sugar release in flasks when combined with 34 mg of protein per 1 gram of organic dry mass. Additionally, another experiment was carried out to investigate the impact of enzymatic pretreatment on biogas production using artificial biowaste as a substrate. Artificial biowaste produced 577,9 NL/kg oDM, while enzymatically pretreated biowaste produced 639,3 NL/kg oDM. This resulted in a 10,6% rise in cumulative biogas production compared to its use without enzymatic pretreatment. By the conclusion of the investigation, specific cumulative dry methane yields of 364,7 NL/kg oDM and 426,3 NL/kg oDM were obtained from artificial biowaste without and with enzymatic pretreatment, respectively. This resulted in a methane production boost of 16,9%. Additionally in case of the reactors with enzymatically pretreated substrate kinetic constant was lower more than double, where maximum volume of biogas increased, comparing to the reactors without enzymatic pretreatment.