Journal Article

A Penicillium rubens platform strain for secondary metabolite production

Polli, Fabiola, Schütze, Tabea, Bovenberg, Roel A. L., de Vries, Maaike, Driessen, Arnold J. M., Meyer, Vera, Pohl, Carsten, Viggiano, Annarita, Mózsik, László, Jung, Sascha

2020

Overview

We present a Penicillium rubens strain with an industrial background in which the four highly expressed biosynthetic gene clusters (BGC) required to produce penicillin, roquefortine, chrysogine and fungisporin were removed. This resulted in a minimal secondary metabolite background. Amino acid pools under steady-state growth conditions showed reduced levels of methionine and increased intracellular aromatic amino acids. Expression profiling of remaining BGC core genes and untargeted mass spectrometry did not identify products from uncharacterized BGCs. This platform strain was repurposed for expression of the recently identified polyketide calbistrin gene cluster and achieved high yields of decumbenone A, B and C. The penicillin BGC could be restored through in vivo assembly with eight DNA segments with short overlaps. Our study paves the way for fast combinatorial assembly and expression of biosynthetic pathways in a fungal strain with low endogenous secondary metabolite burden.

• Publisher: Nature Publishing Group UK,Nature Publishing Group
• Subject: 631/61/191/2018 / 631/61/252/22 / Amino acids / Biosynthetic Pathways - genetics / Gene clusters / Gene Expression Regulation, Enzymologic / Gene Expression Regulation, Fungal / Genome, Fungal / Genomics - methods / Growth conditions / Humanities and Social Sciences / Mass spectrometry / Mass spectroscopy / Metabolic Engineering / Metabolites / Methionine / multidisciplinary / Multigene Family / Penicillin / Penicillium / Penicillium - classification / Penicillium - genetics / Penicillium - metabolism / Peptide Synthases - genetics / Peptide Synthases - metabolism / Science / Science (multidisciplinary) / Secondary Metabolism / Transcriptome