Browsing by Author "Morales Valencia, Karla Pamela"
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Item Determinación del ph y temperatura óptimos de la enzima xilanasa de Trichoderma reesei (TrXynIV) empleando xilano de haya como sustrato(Universidad Técnica de Ambato. Facultad de Ciencia e Ingeniería en Alimentos y Biotecnología. Carrera de Biotecnología, 2024-08) Morales Valencia, Karla Pamela; Cerda Mejía, Liliana AlexandraIn recent years, the global population has doubled, leading to an increased demand for resources and environmental pollution due to the indiscriminate extraction of raw materials. This has generated the need to use more sustainable alternatives, such as lignocellulose, which is one of the most abundant sources of biomass. However, its complex structure requires costly and polluting pretreatments, making it less profitable. Despite this, alternative pretreatments have been developed, such as the use of enzymes capable of degrading lignocellulose. The fungus Trichoderma reesei is known for producing enzymes effective in the decomposition of biomass, such as xylanases from the GH30 family. Among these enzymes, XynVI stands out for its efficiency in lignocellulose degradation. In this project, the activity of xylanase XynVI was analyzed, from plasmid construction to its expression in E. coli BL21 (DE3) and purification by affinity chromatography (IMAC). Additionally, the optimal pH and temperature of the enzyme were evaluated using beechwood xylan as a substrate. As a result, deficient expression of the enzyme and low concentration after purification were observed. Nonetheless, XynVI showed an enzymatic activity of approximately 5 units per milligram of protein at a temperature of 60 degrees Celsius and pH of 5.1, with a difference of 10 degrees above the optimal temperature reported in the literature.