Heterologous expression of the avirulence gene ACE1 from the fungal rice pathogen Magnaporthe oryzae

Song, Zhiyong; Bakeer, Walid; Marshall, James W.; Yakasai, Ahmed A.; Khalid, Rozida Mohd.; Collemare, Jérôme; Skellam, Elizabeth; Tharreau, Didier; Lebrun, Marc; Lazarus, Colin M.; Bailey, Andy M.; Simpson, Thomas J.; Cox, Russell J.

doi:10.1039/c4sc03707c

Cited by 73 publications

(76 citation statements)

References 35 publications

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“…Formation of similar over-reduced products from PKS-NRPKS has been observed. 14 Hence, this suggests that PoxO is responsible for the Knoevenagel condensation. More importantly, the structure of 12, and that inferred for 13, reveal that the aliphatic chain may be derived from incorporation of a nonproteinogenic amino acid, ( S, E )-2-aminodec-4-enoic acid ( 14 ), by the NRPS domain of PoxE (Figure 3).…”

Section: Figurementioning

confidence: 95%

Collaborative Biosynthesis of Maleimide- and Succinimide-Containing Natural Products by Fungal Polyketide Megasynthases

Sato

Dander²,

Sato³

et al. 2017

J. Am. Chem. Soc.

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Fungal polyketide synthases (PKSs) can function collaboratively to synthesize natural products of significant structural diversity. Here we present a new mode of collaboration between a highly reducing PKS (HRPKS) and a PKS-nonribosomal peptide synthetase (PKS-NRPS) in the synthesis of oxaleimides from the Penicillium species. The HRPKS is recruited in the synthesis of an olefin-containing free amino acid, which is activated and incorporated by the adenylation domain of the PKS-NRPS. The precisely positioned olefin from the unnatural amino acid is proposed to facilitate a scaffold rearrangement of the PKS-NRPS product to forge the maleimide and succinimide cores of oxaleimides.

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Section: Figurementioning

confidence: 95%

Collaborative Biosynthesis of Maleimide- and Succinimide-Containing Natural Products by Fungal Polyketide Megasynthases

Sato

Dander²,

Sato³

et al. 2017

J. Am. Chem. Soc.

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“…Further secondary metabolites involved in the virulence of the rice blast fungus were believed to be produced by the polyketide synthase [PKS-nonribosomal peptide synthase (NRPS)] MoAce1p. The gene MoACE1 encodes an intracellular hybrid protein which was found to be involved in effects concerning the rice resistance gene Pi33 [7,8].…”

Section: Introductionmentioning

confidence: 99%

Unravelling the biosynthesis of pyriculol in the rice blast fungus Magnaporthe oryzae

et al. 2017

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Pyriculol was isolated from the rice blast fungus Magnaporthe oryzae and found to induce lesion formation on rice leaves. These findings suggest that it could be involved in virulence. The gene MoPKS19 was identified to encode a polyketide synthase essential for the production of the polyketide pyriculol in the rice blast fungus M. oryzae. The transcript abundance of MoPKS19 correlates with the biosynthesis rate of pyriculol in a time-dependent manner. Furthermore, gene inactivation of MoPKS19 resulted in a mutant unable to produce pyriculol, pyriculariol and their dihydro derivatives. Inactivation of a putative oxidase-encoding gene MoC19OXR1, which was found to be located in the genome close to MoPKS19, resulted in a mutant exclusively producing dihydropyriculol and dihydropyriculariol. By contrast, overexpression of MoC19OXR1 resulted in a mutant strain only producing pyriculol. The MoPKS19 cluster, furthermore, comprises two transcription factors MoC19TRF1 and MoC19TRF2, which were both found individually to act as negative regulators repressing gene expression of MoPKS19. Additionally, extracts of DMopks19 and DMoC19oxr1 made from axenic cultures failed to induce lesions on rice leaves compared to extracts of the wild-type strain. Consequently, pyriculol and its isomer pyriculariol appear to be the only lesioninducing secondary metabolites produced by M. oryzae wild-type (MoWT) under these culture conditions. Interestingly, the mutants unable to produce pyriculol and pyriculariol were as pathogenic as MoWT, demonstrating that pyriculol is not required for infection.

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“…Similar oxidation products 12A and 13A have been reported in the heterologous production in A. oryzae of desmethylbassianin (13) 15 and PKS-NRPS product 13 from Magnaporthe grisea. 16 In all cases including 1 and 2a/2b, oxidations occurred at the terminal of linear polyketide precursors 12 and 13 ( Figure 4). …”

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confidence: 99%

Reconstitution of biosynthetic machinery of fungal polyketides: unexpected oxidations of biosynthetic intermediates by expression host

Fujii

Ugai

Ichinose

et al. 2016

Bioscience, Biotechnology, and Biochemistry

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AbstractsReconstitution of whole biosynthetic genes in Aspergillus oryzae has successfully applied for total biosynthesis of various fungal natural products. Heterologous production of fungal metabolites sometimes suffers unexpected side reactions by host enzymes. In the studies on fungal polyketides solanapyrone and cytochalasin, unexpected oxidations of terminal olefin of biosynthetic intermediates were found to give one and four by-products by host enzymes of the transformants harboring biosynthetic genes. In this paper, we reported structure determination of by-products and described a simple solution to avoid the undesired reaction by introducing the downstream gene in the heterologous production of solanapyrone C. Initial introduction using pTAex3 harboring sol1 in A. oryzae NSAR1 has successfully gave transformants AO-sol1. AO-sol1 produced desmethylprosolanapyrone I (5) in 120 mg/kg whose methylation product (Me 2 SO 4 , acetone) was identical to the synthetic prosolanapyrone I (6) (Figure 2). 14 To the AO-sol1, we introduced the second gene sol2 with a plasmid pPTRI-sol2. The resultant AO-sol1/2 produced 6 and significant amount of by-products 11a-11d (Figure 3). Based on their characteristic UV spectra, we speculated that all of them possessed 2-pyrone moiety derived from 6. Molecular formula C 18 H 26 O 5 of a major product 11a suggested that 11a Keywordshad the same carbon framework of 6. Compared its 1 H NMR spectrum with that of 6, up-field shifted doublet methyl signal (1.13 ppm), and two oxymethine signals (3.85, 4.01 ppm) were newly observed in place of a terminal propenyl moiety in 6, suggesting that the terminal olefin in 6 was oxidized to a vicinal diol in 11a. Structure of 11a was further confirmed by several 2D-NMR data as shown in Figure 3. Similarly, structures of three additional metabolites were determined in a similar manner as shown in the Figure 3. Length of the side chain on the pyrone ring in 11b (C7), 11c (C5) and 11d (C3) suggested that putative C9-product derived by the oxidative cleavage of vic-diol in 11a further degraded by β-oxidation to give 11b-11d. Thus we speculated that host monooxygenase oxidized terminal olefin in 6 to epoxide 10 in which hydrolysis underwent to give diol 11a as in the case of the total biosynthesis of paxilline. 3a Further oxidations were also likely catalyzed by oxidases in A. oryzae. Again, we faced a similar problem which occurred in the cytochalasin project.To avoid the side reaction, we decided to introduce the downstream gene sol6. Our initial attempt to introduce sol6 to the AO-sol1/2 failed to produce prosolanapyrone II (7). 14 When we reexamined DNA sequence of gene cluster with a program 2ndFind which has recently developed for rapid identification of natural product gene cluster, we found the previous prediction of sol6 intron was not correct. 13 Based on this obsevation, we prepared the plasmids pUSA-sol2 and pUSA2-sol2/6 which were subsequently introduced into AO-sol1 to yield AO-sol1/2 and AO-sol1/2/6. We gratifyingly found that the AO-...

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Heterologous expression of the avirulence gene ACE1 from the fungal rice pathogen Magnaporthe oryzae

Abstract: Heterologous expression of key components of the Magnaporthe grisea ACE1 gene cluster produces a potential precursor of cryptic avirulence signalling compounds that induce resistance to M. grisea in rice.

Cited by 73 publications

References 35 publications

Collaborative Biosynthesis of Maleimide- and Succinimide-Containing Natural Products by Fungal Polyketide Megasynthases

Collaborative Biosynthesis of Maleimide- and Succinimide-Containing Natural Products by Fungal Polyketide Megasynthases

Unravelling the biosynthesis of pyriculol in the rice blast fungus Magnaporthe oryzae

Reconstitution of biosynthetic machinery of fungal polyketides: unexpected oxidations of biosynthetic intermediates by expression host

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