Molecular Basis for Mycophenolic Acid Biosynthesis in Penicillium brevicompactum

Regueira, Torsten Ulrik Bak; Kildegaard, Kanchana Rueksomtawin; Hansen, Bertel; Mortensen, Uffe Hasbro; Hertweck, Christian; Nielsen, Jens

doi:10.1128/aem.03015-10

Cited by 139 publications

(144 citation statements)

References 50 publications

(35 reference statements)

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“…A number of compounds from this genus have been characterised and used for drugs and mycotoxins. For example, the antibiotic penicillin is produced by Penicillium rubens (Houbraken et al 2011), the immunosuppressive drug mycophenolic acid is produced by Penicillium brevicompactum (Regueira et al 2011). There were many new compounds that have been found constantly in the following years with impressive anticancer and antifungal activities (Bladt et al 2013; Kozlovskii et al 2013; Tang et al 2015; Koul et al 2016).…”

Section: Introductionmentioning

confidence: 99%

Chemical diversity from the Tibetan Plateau fungi Penicillium kongii and P. brasilianum

Liu

et al. 2017

Mycology

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Two new secondary metabolites, kongiilines A and B (1, 7), and two asperphenamate derivatives, asperphenamates B and C (5–6), together with 16 known compounds (2–4, 8–20), were isolated from Tibetan Plateau fungi Penicillium kongii and Penicillium brasilianum. This is the first report on asperphenamates B and C as naturally occurring compounds, and that aspterric acid is isolated from P. brasilianum for the first time. Their structures were elucidated by different spectroscopic techniques including high-resolution electrospray ionisation mass spectrum, 1D nuclear magnetic resonance (NMR), and 2D NMR as well as electronic circular dichroism. Compounds 4, 5, and 10 exhibited cytotoxicity activities against human colon carcinoma HCT116 cell line with IC50 values of 88.16, 77.68, and 36.92 μM, respectively. Fungi from Tibetan Plateau represent important and rich resources for the investigation of new chemicals.

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Section: Introductionmentioning

confidence: 99%

Chemical diversity from the Tibetan Plateau fungi Penicillium kongii and P. brasilianum

Liu

et al. 2017

Mycology

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“…To demonstrate the potential of the presented USER vector set for heterologus expression and characterization of YFG in A. nidulans, we decided to investigate a recently discovered gene cluster from Penicillium brevicompactum that has been proposed to encode the enzymes required for MPA production (25). The mpaC gene in the cluster consists of 7,745 bp, including introns, and is predicted to encode a polyketide synthase (PKS) that catalyzes production of the polyketide 5-MOA, which is believed to be the first intermediate in MPA biosynthesis (2).…”

Section: Vol 77 2011 Expression and Characterization System For Aspmentioning

confidence: 99%

“…As a proof of concept, the vector set was used to express the heterologous mpaC gene from Penicillium brevicompactum (see the accompanying manuscript in this issue by Regueira et al [25]). mpaC is located in a gene cluster predicted to encode the biosynthetic enzymes for the production of the immunosuppressant mycophenolic acid.…”

mentioning

confidence: 99%

Versatile Enzyme Expression and Characterization System for Aspergillus nidulans, with the Penicillium brevicompactum Polyketide Synthase Gene from the Mycophenolic Acid Gene Cluster as a Test Case

Hansen

Salomonsen

Nielsen

et al. 2011

Appl Environ Microbiol

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105

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Assigning functions to newly discovered genes constitutes one of the major challenges en route to fully exploiting the data becoming available from the genome sequencing initiatives. Heterologous expression in an appropriate host is central in functional genomics studies. In this context, filamentous fungi offer many advantages over bacterial and yeast systems. To facilitate the use of filamentous fungi in functional genomics, we present a versatile cloning system that allows a gene of interest to be expressed from a defined genomic location of Aspergillus nidulans. By a single USER cloning step, genes are easily inserted into a combined targeting-expression cassette ready for rapid integration and analysis. The system comprises a vector set that allows genes to be expressed either from the constitutive PgpdA promoter or from the inducible PalcA promoter. Moreover, by using the vector set, protein variants can easily be made and expressed from the same locus, which is mandatory for proper comparative analyses. Lastly, all individual elements of the vectors can easily be substituted for other similar elements, ensuring the flexibility of the system. We have demonstrated the potential of the system by transferring the 7,745-bp large mpaC gene from Penicillium brevicompactum to A. nidulans. In parallel, we produced defined mutant derivatives of mpaC, and the combined analysis of A. nidulans strains expressing mpaC or mutated mpaC genes unequivocally demonstrated that mpaC indeed encodes a polyketide synthase that produces the first intermediate in the production of the medically important immunosuppressant mycophenolic acid.

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“…Initial annotation of the eight proposed ORFs that constitutes the putative MPA biosynthetic cluster in P. brevicompactum suggested the following candidates for the established biosynthetic reactions: (i) polyketide synthase (PKS), mpaC; (ii) cytochrome P450, mpaD; (iii) prenyl transferase, mpaA; (iv) oxidative cleaving, mpaH; (v) and finally O-methyltransferase, mpaG (23). However, the lactonization occurring at the second step in MPA biosynthesis is not a standard cytochrome P450 reaction, which prompted us to investigate this step in detail.…”

Section: Resultsmentioning

confidence: 99%

“…1A). However, the biosynthesis remained unelucidated at the genetic level until the recent discovery of a putative MPA biosynthetic cluster in Penicillium brevicompactum (23). The defined cluster contained eight putative open reading frames (ORFs), including mpaC, which encodes a polyketide synthase catalyzing the production of 5-methylorsellinic acid (5-MOA), the first step in MPA biosynthesis (9,23).…”

mentioning

confidence: 99%

Involvement of a Natural Fusion of a Cytochrome P450 and a Hydrolase in Mycophenolic Acid Biosynthesis

Hansen

Mnich

Nielsen

et al. 2012

Appl Environ Microbiol

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Mycophenolic acid (MPA) is a fungal secondary metabolite and the active component in several immunosuppressive pharmaceuticals. The gene cluster coding for the MPA biosynthetic pathway has recently been discovered in Penicillium brevicompactum, demonstrating that the first step is catalyzed by MpaC, a polyketide synthase producing 5-methylorsellinic acid (5-MOA). However, the biochemical role of the enzymes encoded by the remaining genes in the MPA gene cluster is still unknown. Based on bioinformatic analysis of the MPA gene cluster, we hypothesized that the step following 5-MOA production in the pathway is carried out by a natural fusion enzyme MpaDE, consisting of a cytochrome P450 (MpaD) in the N-terminal region and a hydrolase (MpaE) in the C-terminal region. We verified that the fusion gene is indeed expressed in P. brevicompactum by obtaining full-length sequence of the mpaDE cDNA prepared from the extracted RNA. Heterologous coexpression of mpaC and the fusion gene mpaDE in the MPA-nonproducer Aspergillus nidulans resulted in the production of 5,7-dihydroxy-4-methylphthalide (DHMP), the second intermediate in MPA biosynthesis. Analysis of the strain coexpressing mpaC and the mpaD part of mpaDE shows that the P450 catalyzes hydroxylation of 5-MOA to 4,6-dihydroxy-2-(hydroxymethyl)-3-methylbenzoic acid (DHMB). DHMB is then converted to DHMP, and our results suggest that the hydrolase domain aids this second step by acting as a lactone synthase that catalyzes the ring closure. Overall, the chimeric enzyme MpaDE provides insight into the genetic organization of the MPA biosynthesis pathway.

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Molecular Basis for Mycophenolic Acid Biosynthesis in Penicillium brevicompactum

Cited by 139 publications

References 50 publications

Chemical diversity from the Tibetan Plateau fungi Penicillium kongii and P. brasilianum

Chemical diversity from the Tibetan Plateau fungi Penicillium kongii and P. brasilianum

Versatile Enzyme Expression and Characterization System for Aspergillus nidulans, with the Penicillium brevicompactum Polyketide Synthase Gene from the Mycophenolic Acid Gene Cluster as a Test Case

Involvement of a Natural Fusion of a Cytochrome P450 and a Hydrolase in Mycophenolic Acid Biosynthesis

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