Получение промышленно важных ферментов на основе экспрессионной системы гриба Penicillium verruculosum

Abstract

The review provides the data on the creation of highly active recombinant strains producers of a wide range of enzymes for various fields of industry and agriculture using the expression system of the Penicillium verruculosum filamentous fungus: endoglucanases, β-glucanases, xylanases, mannanases, phytases, α-galactosidases, pectin lyases, polygalacturonases, dextranases, exo-inulinases and chitinases. The recipient strain, P. verruculosum B1-537(ΔniaD), has a high secretory capacity (up to 60 g/L of extracellular protein in the culture liquid), and the recombinant producer strains are characterized by high productivity of target enzymes. The obtained enzyme preparations contain, as a rule, 30--70% of the target recombinant enzymes (in some cases, up to 80%) of the total protein pool. The P. verruculosum expression system makes it easy to transform expression constructs containing target heterologous or homologous genes, functionally linked to the promoter and terminator of the strong inducible promoter of the cellobiohydrolase-1 (cbh1) gene, and the genetic transformation of the recipient strain can be carried out simultaneously using several vector constructs. The incorporation of target genes into the chromosome of the recipient strain is an integrative process, and the foreign recombinant proteins encoded by them are not subjected to proteolysis. The technique of primary selection of positive transformants on agar media, as well as secondary selection during their cultivation in shaking flasks and fermenters, is simple and reliable in use, partly owing to standardized parameters: the nutrient composition of the fermentation medium and the culturing conditions of recombinant strains producing various target enzymes. The time spent on creating a producer of the target enzyme, from setting the problem to obtaining a recombinant strain, takes from 3 to 6 months. It is important to emphasize that recombinant P. verruculosum strains are successfully used in industrial processes as producers of various target enzymes. expression system, cloning, industrial enzymes, feed additives, Penicillium verruculosum The work was supported by the Ministry of Science and Higher Education of the Russian Federation (State Assignment 0104-2019-0009).