Genome organization and assessment of high copy number and increased expression of pectinolytic genes from Penicillium griseoroseum: a potential heterologous system for protein production

• Published in: Journal of industrial microbiology & biotechnology Vol. 41; no. 10; pp. 1571 - 1580
• Main Authors: Teixeira, Janaina Aparecida, Nogueira, Guilherme Bicalho, de Queiroz, Marisa Vieira, de Araújo, Elza Fernandes
• Format: Journal Article
• Language: English
• Published: Berlin/Heidelberg Springer-Verlag 01-10-2014; Springer Berlin Heidelberg; Springer; Oxford University Press
• Subjects: Analysis; Aspergillus nidulans; Aspergillus nidulans - genetics; Bacteria; Base Sequence; Biochemistry; Bioinformatics; Biological and medical sciences; Biomedical and Life Sciences; Biotechnology; Chromosomes; Enzymes; Fundamental and applied biological sciences. Psychology; Fungal Proteins - biosynthesis; Fungal Proteins - genetics; Fungi; Gene Dosage; Gene Expression; Gene Expression Regulation, Fungal; genes; Genetic Engineering; Genetics and Molecular Biology of Industrial Organisms; Genome, Fungal; Genomes; Growth conditions; industrial applications; Inorganic Chemistry; Life Sciences; Microbiology; Mycotoxins; Penicillin; Penicillium; Penicillium - enzymology; Penicillium - genetics; polygalacturonase; Polygalacturonase - biosynthesis; Polygalacturonase - genetics; Polysaccharide-Lyases - biosynthesis; Polysaccharide-Lyases - genetics; Promoter Regions, Genetic; Protein Biosynthesis; protein secretion; Protein synthesis; proteinases; Proteins; Studies; transcription (genetics); Transcription, Genetic; translation (genetics); PFGE; Genome organization; Protein production; Gene expression; Pectinase; Enzyme; Copy number; Penicillium; Polygalacturonase; Fungi; Glycosylases; Gene; Glycosidases; Production; Penicillium griseoroseum; Hydrolases; Fungi Imperfecti; Recombinant protein
• DOI: 10.1007/s10295-014-1486-4
• EISSN: 1476-5535
• ISSN: 1367-5435

The fungus Penicillium griseoroseum has the potential for application on an industrial scale as a host for the production of homologous and heterologous proteins, mainly because it does not produce some mycotoxins or secrete proteases under the growth conditions for pectinase production. However, for the fungus to be used effectively as an expression heterologous system, an understanding of the organization of its genome, as well as the mechanisms of gene expression and protein production, is required. In the present study, the size of the P. griseoroseum genome was estimated to be 29.8–31.5 Mb, distributed among four chromosomes. An analysis of plg1 and pgg2 pectinolytic genes expression and copy number in recombinant multi-copy strains of P. griseoroseum demonstrated that an increase in the number of gene copies could increase enzyme production, but the transcription could be affected by the gene integration position. Placing a copy of the plg1 gene under the control of the gpd promoter of Aspergillus nidulans yielded a 200-fold increase in transcription levels compared to the endogenous gene, and two copies of the pgg2 gene produced an 1100-fold increase compared with the endogenous gene. These results demonstrated that transcription, translation, and protein secretion in the fungus P. griseoroseum respond to an increased number of gene copies in the genome. The processing capacity and efficiency of protein secretion in P. griseoroseum are consistent with our premise that this fungus can be used for the industrial-scale production of several enzymes.