Construction and Identification of a Food-Grade Recombinant Cyberlindnera jadinii Strain Expressing Lignin Peroxidase

Abstract

Lignin peroxidase is one of a series of enzymes involved in lignin degradation and is secreted by white-rot fungi. However, white-rot fungi have a long growth cycle and low enzyme activity, which limits their application. An efficient and stable heterologous expression system might be the solution to overcome these critical limitations. In this study, the lignin peroxidase gene of white-rot fungus (Phanerodontia chrysosporium) was cloned and heterologously expressed in the food-grade yeast Cyberlindnera jadinii for the first time. The strain ZHMX4 expressing lignin peroxidase was constructed, and its genetic stability, enzyme activity and phenotypic analysis were evaluated. ZHMX4 retained and maintained genetic stability for up to 100 generations. Its lignin peroxidase activity reached a maximum at 96 h with 68.52 U/L. Phenotypic analysis (Biolog) showed obvious changes in the substrate metabolism of the engineered strain compared to Cyberlindnera jadinii. Thus, ZHMX4 is a food-grade genetically engineered strain with great potential for lignin degradation. This study successfully expressed lignin peroxidase in Cyberlindnera jadinii as a new approach for the biodegradation of lignin. These findings should be useful for further academic studies, industrial applications of lignin peroxidase and construction of artificial white-rot fungi.