Lipolytic production from solid-state fermentation of the filamentous fungus Penicillium polonicum and its applicability as biocatalyst in the synthesis of ethyl oleate.

Lipases represent versatile biocatalysts extensively employed in transesterification reactions for ester production. Ethyl oleate holds significance in biodiesel production, serving as a sustainable alternative to petroleum-derived diesel. In this study, our goal was to prospect lipase and assess its efficacy as a biocatalyst for ethyl oleate synthesis. For quantitative analysis, a base medium supplemented with Rhodamine B, olive oil, and Tween 80 was used. Solid-state fermentation utilized crambe seeds of varying particle sizes and humidity levels as substrates. In the synthesis of ethyl oleate, molar ratios of 1:3, 1:6, and 1:9, along with a total enzymatic activity of 60 U in n-heptane, were utilized at temperatures of 30 °C, 37 °C, and 44 °C. Reactions were conducted in a shaker at 200 rpm for 60 min. As a result, we first identified Penicillium polonicum and employed the method of solid-state fermentation using crambe seeds as a substrate to produce lipase. Our findings revealed heightened lipolytic activity (22.5 Ug-1) after 96 h of fermentation using crambe cake as the substrate. Optimal results were achieved with crambe seeds at a granulometry of 0.6 mm and a fermentation medium humidity of 60%. Additionally, electron microscopy suggested the immobilization of lipase in the substrate, enabling enzyme reuse for up to 4 cycles with 100% enzymatic activity. Subsequently, we conducted applicability tests of biocatalysts for ethyl oleate synthesis, optimizing parameters such as the acid/alcohol molar ratio, temperature, and reaction time. We attained 100% conversion within 30 min at 37 °C, and our results indicated that the molar ratio proportion did not significantly influence the outcome. These findings provide a methodological alternative for the utilization of biocatalysts in ethyl oleate synthesis.

Aspergillus sclerotiorum lipolytic activity and its application in bioremediation of high-fat dairy wastewater environments.

Oils and grease (O&G) have low affinity for water and represent a class of pollutants present in the dairy industry. Enzyme-mediated bioremediation using biocatalysts, such as lipases, has shown promising potential in biotechnology, as they are versatile catalysts with high enantioselectivity and regioselectivity and easy availability, being considered a clean technology (white biotechnology). Specially in the treatment of effluents from dairy industries, these enzymes are of particular importance as they specifically hydrolyze O&G. In this context, the objective of this work is to prospect filamentous fungi with the ability to synthesize lipases for application in a high-fat dairy wastewater environment. We identified and characterized the fungal species Aspergillus sclerotiorum as a good lipase producer. Specifically, we observed highest lipolytic activity (20.72 U g-1) after 96 h of fermentation using sunflower seed as substrate. The fungal solid fermented was used in the bioremediation in dairy effluent to reduce O&G. The experiment was done in kinetic from 24 to 168 h and reduced over 90% of the O&G present in the sample after 168 h. Collectively, our work demonstrated the efficiency and applicability of fungal fermented solids in bioremediation and how this process can contribute to a more sustainable wastewater pretreatment, reducing the generation of effluents produced by dairy industries.