ArticleFrontiers in microbiology2015
A new high-performance heterologous fungal expression system based on regulatory elements from the Aspergillus terreus terrein gene cluster.
Article in Frontiers in microbiology, 2015. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 37 papers.
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37 citing papers in PubMed, 125 citations in OpenAlex.
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- Total Heterologous Biosynthesis of Fungal Natural Products inJournal of natural products · 2022Review
- Enhanced production of terrein in marine-derived Aspergillus terreus by refactoring both global and pathway-specific transcription factors.Microbial cell factories · 2022Article
- Unprecedented Mushroom Polyketide Synthases Produce the Universal Anthraquinone Precursor.Angewandte Chemie (International ed. in English) · 2022Article
- Characterisation of ascocorynin biosynthesis in the purple jellydisc fungus Ascocoryne sarcoides.Fungal biology and biotechnology · 2022Article
- Construction of a novel filamentous fungal protein expression system based on redesigning of regulatory elements.Applied microbiology and biotechnology · 2022Article
- Identification of a lichen depside polyketide synthase gene by heterologous expression inMetabolic engineering communications · 2021Article
- Early Oxidative Transformations During the Biosynthesis of Terrein and Related Natural Products.Chemistry (Weinheim an der Bergstrasse, Germany) · 2021Article
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4 authors at 1 institution in 1 country.
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No grant is acknowledged in the PubMed record.
Abstract
Recently, the Aspergillus terreus terrein gene cluster was identified and selected for development of a new heterologous expression system. The cluster encodes the specific transcription factor TerR that is indispensable for terrein cluster induction. To identify TerR binding sites, different recombinant versions of the TerR DNA-binding domain were analyzed for specific motif recognition. The high affinity consensus motif TCGGHHWYHCGGH was identified from genes required for terrein production and binding site mutations confirmed their essential contribution to gene expression in A. terreus. A combination of TerR with its terA target promoter was tested as recombinant expression system in the heterologous host Aspergillus niger. TerR mediated target promoter activation was directly dependent on its transcription level. Therefore, terR was expressed under control of the regulatable amylase promoter PamyB and the resulting activation of the terA target promoter was compared with activation levels obtained from direct expression of reporters from the strong gpdA control promoter. Here, the coupled system outcompeted the direct expression system. When the coupled system was used for heterologous polyketide synthase expression high metabolite levels were produced. Additionally, expression of the Aspergillus nidulans polyketide synthase gene orsA revealed lecanoric acid rather than orsellinic acid as major polyketide synthase product. Domain swapping experiments assigned this depside formation from orsellinic acid to the OrsA thioesterase domain. These experiments confirm the suitability of the expression system especially for high-level metabolite production in heterologous hosts.
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