title: Genome organization and assessment of high copy number and increased expression of pectinolytic genes from Penicillium griseoroseum: a potential heterologous system for protein production.
abstract: 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.
authors.affiliation: Departamento de Microbiologia/BIOAGRO, Universidade Federal de Viçosa, Viçosa, MG, CEP 36570-000, Brazil.
authors.forename: Janaina Aparecida
authors.initials: JA
authors.lastname: Teixeira
authors.forename: Guilherme Bicalho
authors.initials: GB
authors.lastname: Nogueira
authors.forename: Marisa Vieira
authors.initials: MV
authors.lastname: de Queiroz
authors.forename: Elza Fernandes
authors.initials: EF
authors.lastname: de Araújo
mesh_terms: D001233:Aspergillus nidulans / Q000235:genetics; D001483:Base Sequence; D005656:Fungal Proteins / Q000096:biosynthesis / Q000235:genetics*; D018628:Gene Dosage; D015870:Gene Expression; D015966:Gene Expression Regulation, Fungal; D016681:Genome, Fungal; D010407:Penicillium / Q000201:enzymology / Q000235:genetics*; D011096:Polygalacturonase / Q000096:biosynthesis / Q000235:genetics*; D011133:Polysaccharide-Lyases / Q000096:biosynthesis / Q000235:genetics*; D011401:Promoter Regions, Genetic; D014176:Protein Biosynthesis; D014158:Transcription, Genetic
chemical_list: D005656:Fungal Proteins; D011096:Polygalacturonase; D011133:Polysaccharide-Lyases; C027349:pectin lyase
references.citation: Can J Microbiol. 2006 Nov;52(11):1070-7
references.pmid: 17215898
references.citation: J Appl Microbiol. 2011 Oct;111(4):818-25
references.pmid: 21726361
references.citation: Can J Microbiol. 2004 Nov;50(11):891-900
references.pmid: 15644906
references.citation: Genet Mol Biol. 2009 Jan;32(1):129-32
references.pmid: 21637657
references.citation: FEMS Yeast Res. 2009 Dec;9(8):1260-70
references.pmid: 19799640
references.citation: J Microbiol Biotechnol. 2012 Aug;22(8):1133-40
references.pmid: 22713991
references.citation: Can J Microbiol. 2010 Oct;56(10):831-7
references.pmid: 20962906
references.citation: Fungal Genet Biol. 2002 Feb;35(1):21-9
references.pmid: 11860262
references.citation: Trends Biotechnol. 2005 Sep;23(9):468-74
references.pmid: 15967521
references.citation: Nucleic Acids Res. 2001 May 1;29(9):e45
references.pmid: 11328886
references.citation: Mol Microbiol. 2001 Jun;40(6):1345-56
references.pmid: 11442833
references.citation: Methods. 2001 Dec;25(4):402-8
references.pmid: 11846609
references.citation: J Ind Microbiol Biotechnol. 2008 Mar;35(3):159-66
references.pmid: 18030511
references.citation: J Immunol Methods. 2003 Jul;278(1-2):261-9
references.pmid: 12957413
references.citation: J Appl Microbiol. 2008 Nov;105(5):1595-603
references.pmid: 19146495
references.citation: Int J Biol Sci. 2010 Feb 06;6(1):96-106
references.pmid: 20150979
references.citation: Int J Food Microbiol. 2000 Jun 30;58(1-2):59-63
references.pmid: 10898462
references.citation: Microbiol Rev. 1993 Jun;57(2):383-401
references.pmid: 8393130
references.citation: Nat Biotechnol. 2008 Oct;26(10):1161-8
references.pmid: 18820685
references.citation: Mol Gen Genet. 1993 Dec;241(5-6):573-8
references.pmid: 8264531
references.citation: FEMS Microbiol Lett. 1997 Dec 15;157(2):267-72
references.pmid: 9435107
references.citation: Anal Biochem. 1982 Jan 1;119(1):158-63
references.pmid: 7041693
references.citation: Mol Cell Biol. 1988 Dec;8(12):5513-20
references.pmid: 2854200
references.citation: Appl Biochem Biotechnol. 2013 Mar;169(6):1965-77
references.pmid: 23354503
references.citation: J Basic Microbiol. 2014 Feb;54(2):133-41
references.pmid: 23553535
references.citation: Nucleic Acids Res. 2003 May 15;31(10):e56
references.pmid: 12736322
references.citation: Arch Biochem Biophys. 1962 Apr;97:107-15
references.pmid: 13860094