Co-expression of β-Glucanase Genes of Various Origin as an Approach to the Creation of Recombinant Yeast Strains Producing Enzymes with Optimal Properties

Abstract

The co-expression of genes encoding β-glucanases from Paenibacillus jamilae and Rhizomucor miehei in the Komagataella kurtzmanii methylotrophic yeast system has been carried out. The recombinant strains that simultaneously produced both enzymes with a total activity of 1132 and 1464 units for 96 h of cultivation in flasks were obtained. The combination of enzyme properties made it possible to achieve a substantial extension of the pH range of β-glucanase activity, as well as to bring the temperature and pH optima of the enzymes closer to those optimal for use in feed production. A vector has been developed that allows the efficient simultaneous expression of genes due to the sequential integration of expression cassettes into the recipient strain chromosome. It was also shown that a change in the ratio of the number of genes integrated into the chromosome of the host cell allows to directionally influence the total properties of recombinant enzymes. co-expression, β-glucanase, β-glucan, Paenibacillus jamilae, Rhizomucor miehei, Komagataella kurtzmanii, Pichia pastoris The work was financially supported by the Ministry of Science and Education (Project Unique Identifier RFMEFI60717X0179) using the Multipurpose Scientific Installation of National Bio-Resource Center «All-Russian Collection of Industrial Microorganisms», NRC «Kurchatov Institute» - GOSNIIGENETIKA.