The heterologous expression of the laccase enzyme from H. tenuis

Abstract

Currently, there is a demand for alternative sources that reduce and/or control the effects of widespread pollution worldwide. Second generation ethanol is a biofuel that can be produced through the hydrolysis process of polysaccharides using lignocellulosic biomass with the addition of enzymatic complexes. For the conversion of lignocellulosic biomass into ethanol, this material must undergo the hydrolysis process to convert it into fermentable sugars. However, the enzymes currently used are costly, which limits the process. In this regard, the search for enzymes that can optimize second generation ethanol production is of paramount importance. Lignin is a polysaccharide constituent of biomass that can also be applied in the production of energy sources. Its application is underexplored and commercially underutilized due to its resistance to enzymatic and chemical hydrolysis, primarily due to its cross-linking with cellulose and other constituents of the cell wall. Laccases are a group of ligninolytic enzymes widely used for this purpose and have been the subject of numerous biotechnological studies in recent years. They have been employed in various industrial sectors, from the textile industry to the juice industry. Additionally, they are enzymes found in diverse organisms, from plants to insects. Among insects, termites are recognized for their high capacity for lignocellulose digestion. Therefore, this project aims to promote the cloning and heterologous expression in Pichia pastoris of a laccase identified in the termite species H. tenuis for possible use in second generation ethanol production. As a result of the project, in addition to its production, tests are expected to be conducted to establish enzymatic activity, determine the ideal pH and temperature for subsequent purification and utilization of this enzyme in the preparation of enzyme cocktails for second generation ethanol production.