title: New promoters for strain engineering of Penicillium chrysogenum.
abstract: Filamentous fungi such as Aspergillus and Penicillium are widely used as hosts for the industrial products such as proteins and secondary metabolites. Although filamentous fungi are versatile in recognizing transcriptional and translational elements present in genes from other filamentous fungal species, only few promoters have been applied and compared in performance so far in Penicillium chrysogenum. Therefore, a set of homologous and heterologous promoters were tested in a reporter system to obtain a set of potential different strengths. Through in vivo homologous recombination in Saccharomyces cerevisiae, twelve Aspergillus niger and P. chrysogenum promoter-reporter pathways were constructed that drive the expression of green fluorescent protein while concurrent expression of the red fluorescent protein was used as an internal standard and placed under control of the PcPAF promoter. The pathways were integrated into the genome of P. chrysogenum and tested using the BioLector system for fermentation. Reporter gene expression was monitored during growth and classified according to promoter strength and expression profile. A set of novel promoters was obtained that can be used to tune the expression of target genes in future strain engineering programs.
authors.affiliation: Molecular Microbiology, Groningen Biomolecular Sciences and Biotechnology Institute, University of Groningen, Groningen, The Netherlands.
authors.forename: Fabiola
authors.initials: F
authors.lastname: Polli
authors.affiliation: DSM Biotechnology Centre, Delft, The Netherlands.
authors.forename: Ben
authors.initials: B
authors.lastname: Meijrink
authors.affiliation: DSM Biotechnology Centre, Delft, The Netherlands; Synthetic Biology and Cell Engineering, Groningen Biomolecular Sciences and Biotechnology Institute, University of Groningen, Groningen, The Netherlands.
authors.forename: Roel A L
authors.initials: RAL
authors.lastname: Bovenberg
authors.affiliation: Molecular Microbiology, Groningen Biomolecular Sciences and Biotechnology Institute, University of Groningen, Groningen, The Netherlands; Kluyver Centre for Genomics of Industrial Fermentations, Julianalaan 67, 2628BC Delft, The Netherlands. Electronic address: a.j.m.driessen@rug.nl.
authors.forename: Arnold J M
authors.initials: AJM
authors.lastname: Driessen
mesh_terms: D000595:Amino Acid Sequence; D001234:Aspergillus niger / Q000235:genetics; D005285:Fermentation; D015870:Gene Expression; D005800:Genes, Fungal; D017930:Genes, Reporter; D005818:Genetic Engineering; D016681:Genome, Fungal; D010408:Penicillium chrysogenum / Q000235:genetics*; D011401:Promoter Regions, Genetic; D012441:Saccharomyces cerevisiae / Q000235:genetics
keywords: Aspergillus niger; Fermentation; Penicillium chrysogenum; Promoters