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Construction of Penicillium crustosum as an expression host for fungal secondary metabolite biosynthesis

Affiliation
Institut für Pharmazeutische Biologie und Biotechnologie, Fachbereich Pharmazie, Philipps-Universität Marburg, 35037 Marburg, Germany
Zhou, Jenny;
Affiliation
Institut für Pharmazeutische Biologie und Biotechnologie, Fachbereich Pharmazie, Philipps-Universität Marburg, 35037 Marburg, Germany
Chen, Xiaoling;
Affiliation
Institut für Pharmazeutische Biologie und Biotechnologie, Fachbereich Pharmazie, Philipps-Universität Marburg, 35037 Marburg, Germany
Li, Shu-Ming

Heterologous expression of (silent/cryptic) genes and biosynthetic gene clusters (BGCs) has been proven to be one of the most promising strategies for the discovery of fungal secondary metabolites. However, successful transformation and expression of BGCs are often hindered by host-dependent factors, such as a high secondary metabolite background or low gene targeting efficiencies. Penicillium crustosum PRB-2 is a fast-growing fungus, which predominantly produces terrestric acid, clavatol, and their derivatives peniphenones and penilactones. To construct P. crustosum as an expression host with a clean secondary metabolite background, the two highly expressed BGCs for the production of terrestric acid and clavatol were inactivated. Furthermore, genetic manipulation in P. crustosum was facilitated by deletion of ligD to enhance gene targeting, and by deletion of pyrG and pcribo to expand the repertoire of selection markers. Finally, the commonly used Aspergillus nidulans wA integration site, consisting of the wA pigment gene and its flanking regions, was chosen for replacement of the P. crustosum pigment gene pcr4401.5 Hence, constructs designed for heterologous expression in A. nidulans can be also used for P. crustosum and transformants can be easily identified by their albino phenotypes.

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