scholarly journals Biosynthesis of Fusarubins Accounts for Pigmentation of Fusarium fujikuroi Perithecia

2012 ◽  
Vol 78 (12) ◽  
pp. 4468-4480 ◽  
Author(s):  
Lena Studt ◽  
Philipp Wiemann ◽  
Karin Kleigrewe ◽  
Hans-Ulrich Humpf ◽  
Bettina Tudzynski
Keyword(s):  
Secondary Metabolites ◽  
Gene Cluster ◽  
Biosynthetic Pathway ◽  
Polyketide Synthase ◽  
Biological Function ◽  
Fusarium Fujikuroi ◽  
Diverse Group ◽  
Fruiting Bodies ◽  
Content Type ◽  
Encoding Gene

ABSTRACTFusarium fujikuroiproduces a variety of secondary metabolites, of which polyketides form the most diverse group. Among these are the highly pigmented naphthoquinones, which have been shown to possess different functional properties for the fungus. A group of naphthoquinones, polyketides related to fusarubin, were identified inFusariumspp. more than 60 years ago, but neither the genes responsible for their formation nor their biological function has been discovered to date. In addition, although it is known that the sexual fruiting bodies in which the progeny of the fungus develops are darkly colored by a polyketide synthase (PKS)-derived pigment, the structure of this pigment has never been elucidated. Here we present data that link the fusarubin-type polyketides to a defined gene cluster, which we designatefsr, and demonstrate that the fusarubins are the pigments responsible for the coloration of the perithecia. We studied their regulation and the function of the single genes within the cluster by a combination of gene replacements and overexpression of the PKS-encoding gene, and we present a model for the biosynthetic pathway of the fusarubins based on these data.

scholarly journals Genome-Based Cluster Deletion Reveals an Endocrocin Biosynthetic Pathway in Aspergillus fumigatus

2012 ◽  
Vol 78 (12) ◽  
pp. 4117-4125 ◽  
Author(s):  
Fang Yun Lim ◽  
Yanpeng Hou ◽  
Yiming Chen ◽  
Jee-Hwan Oh ◽  
Inhyung Lee ◽  
...  
Keyword(s):  
Aspergillus Fumigatus ◽  
Gene Cluster ◽  
Biosynthetic Pathway ◽  
Polyketide Synthase ◽  
Wild Type Strain ◽  
Global Regulator ◽  
Content Type ◽  
In The Wild ◽  
Secondary Metabolite Gene Cluster

ABSTRACTEndocrocin is a simple anthraquinone frequently identified in extracts of numerous fungi. Several biosynthetic schemes for endocrocin synthesis have been hypothesized, but to date, no dedicated secondary metabolite gene cluster that produces this polyketide as its major metabolite has been identified. Here we describe our biosynthetic and regulatory characterization of the endocrocin gene cluster inAspergillus fumigatus. This is the first report of this anthraquinone in this species. The biosynthetic genes required for endocrocin production are regulated by the global regulator of secondary metabolism, LaeA, and encode an iterative nonreducing polyketide synthase (encA), a physically discrete metallo-β-lactamase type thioesterase (encB), and a monooxygenase (encC). Interestingly, the deletion of a gene immediately adjacent toencC, termedencDand encoding a putative 2-oxoglutarate-Fe(II) type oxidoreductase, resulted in higher levels of endocrocin production than in the wild-type strain, whereas overexpression ofencDeliminated endocrocin accumulation. We found that overexpression of theencAtranscript resulted in higher transcript levels ofencA-Dand higher production of endocrocin. We discuss a model of theenccluster as one evolutionary origin of fungal anthraquinones derived from a nonreducing polyketide synthase and a discrete metallo-β-lactamase-type thioesterase.

scholarly journals The fungal gene cluster for biosynthesis of the antibacterial agent viriditoxin

2019 ◽  
Author(s):  
Andrew S. Urquhart ◽  
Jinyu Hu ◽  
Yit-Heng Chooi ◽  
Alexander Idnurm
Keyword(s):  
Secondary Metabolites ◽  
Gene Cluster ◽  
Gene Disruption ◽  
Biosynthetic Pathway ◽  
Fluorescent Protein ◽  
Model Species ◽  
Paecilomyces Variotii ◽  
Bioinformatic Approaches ◽  
Green Fluorescent ◽  
Fungal Gene

AbstractBackgroundViriditoxin is one of the ‘classical’ secondary metabolites produced by fungi and that has antibacterial and other activities; however, the mechanism of its biosynthesis has remained unknown.ResultsHere, a gene cluster responsible for its synthesis was identified, using bioinformatic approaches from two species that produce viriditoxin and then through gene disruption and metabolite profiling. All eight genes in the cluster inPaecilomyces variotiiwere mutated, revealing their roles in the synthesis of this molecule and establishing its biosynthetic pathway which includes an interesting Baeyer-Villiger monooxygenase catalyzed reaction. Additionally, a candidate catalytically-inactive hydrolase was identified as being required for the stereoselective biosynthesis of (M)-viriditoxin. The localization of two proteins were assessed by fusing these proteins to green fluorescent protein, revealing that at least two intracellular structures are involved in the compartmentalization of the synthesis steps of this metabolite.ConclusionsThe full pathway for synthesis of viriditoxin was established by a combination of genomics, bioinformatics, gene disruption and chemical analysis processes. Hence, this work reveals the basis for the synthesis of an understudied class of fungal secondary metabolites and provides a new model species for understanding the synthesis of biaryl compounds with a chiral axis.

scholarly journals Genetic Manipulation of the Fusarium fujikuroi Fusarin Gene Cluster Yields Insight into the Complex Regulation and Fusarin Biosynthetic Pathway

2013 ◽  
Vol 20 (8) ◽  
pp. 1055-1066 ◽  
Author(s):  
Eva-Maria Niehaus ◽  
Karin Kleigrewe ◽  
Philipp Wiemann ◽  
Lena Studt ◽  
Christian M.K. Sieber ◽  
...  
Keyword(s):  
Gene Cluster ◽  
Biosynthetic Pathway ◽  
Genetic Manipulation ◽  
Fusarium Fujikuroi ◽  
Complex Regulation ◽  
Insight Into

scholarly journals Gut Symbionts from Distinct Hosts Exhibit Genotoxic Activity via Divergent Colibactin Biosynthesis Pathways

2014 ◽  
Vol 81 (4) ◽  
pp. 1502-1512 ◽  
Author(s):  
Philipp Engel ◽  
Maria I. Vizcaino ◽  
Jason M. Crawford
Keyword(s):  
Dna Damage ◽  
Secondary Metabolites ◽  
Biosynthetic Pathway ◽  
Tumor Formation ◽  
Microbial Interactions ◽  
Eukaryotic Cells ◽  
Dependent Manner ◽  
Bacterial Symbionts ◽  
Content Type ◽  
E Coli

ABSTRACTSecondary metabolites produced by nonribosomal peptide synthetase (NRPS) or polyketide synthase (PKS) pathways are chemical mediators of microbial interactions in diverse environments. However, little is known about their distribution, evolution, and functional roles in bacterial symbionts associated with animals. A prominent example is colibactin, a largely unknown family of secondary metabolites produced byEscherichia colivia a hybrid NRPS-PKS biosynthetic pathway that inflicts DNA damage upon eukaryotic cells and contributes to colorectal cancer and tumor formation in the mammalian gut. Thus far, homologs of this pathway have only been found in closely relatedEnterobacteriaceae, while a divergent variant of this gene cluster was recently discovered in a marine alphaproteobacterialPseudovibriostrain. Herein, we sequenced the genome ofFrischella perraraPEB0191, a bacterial gut symbiont of honey bees and identified a homologous colibactin biosynthetic pathway related to those found inEnterobacteriaceae. We show that the colibactin genomic island (GI) has conserved gene synteny and biosynthetic module architecture acrossF. perrara,Enterobacteriaceae, and thePseudovibriostrain. Comparative metabolomics analyses ofF. perraraandE. colifurther reveal that these two bacteria produce related colibactin pathway-dependent metabolites. Finally, we demonstrate thatF. perrara, likeE. coli, causes DNA damage in eukaryotic cellsin vitroin a colibactin pathway-dependent manner. Together, these results support that divergent variants of the colibactin biosynthetic pathway are widely distributed among bacterial symbionts, producing related secondary metabolites and likely endowing its producer with functional capabilities important for diverse symbiotic associations.

scholarly journals Biosynthesis of a Complex Yersiniabactin-Like Natural Product via themicLocus in Phytopathogen Ralstonia solanacearum

2011 ◽  
Vol 77 (17) ◽  
pp. 6117-6124 ◽  
Author(s):  
Martin F. Kreutzer ◽  
Hirokazu Kage ◽  
Peter Gebhardt ◽  
Barbara Wackler ◽  
Hans P. Saluz ◽  
...  
Keyword(s):  
Natural Product ◽  
Gene Cluster ◽  
Ralstonia Solanacearum ◽  
Insertional Mutagenesis ◽  
Polyketide Synthase ◽  
Genome Mining ◽  
Content Type ◽  
A Genome ◽  
Iron Deficient ◽  
Peptide Synthetase

ABSTRACTA genome mining study in the plant pathogenic bacteriumRalstonia solanacearumGMI1000 unveiled a polyketide synthase/nonribosomal peptide synthetase gene cluster putatively involved in siderophore biosynthesis. Insertional mutagenesis confirmed the respective locus to be operational under iron-deficient conditions and spurred the isolation of the associated natural product. Bioinformatic analyses of the gene cluster facilitated the structural characterization of this compound, which was subsequently identified as the antimycoplasma agent micacocidin. The metal-chelating properties of micacocidin were evaluated in competition experiments, and the cellular uptake of gallium-micacocidin complexes was demonstrated inR. solanacearumGMI1000, indicating a possible siderophore role. Comparative genomics revealed a conservation of the micacocidin gene cluster in defined, but globally dispersed phylotypes ofR. solanacearum.

scholarly journals Pathway for the Biosynthesis of the Pigment Chrysogine by Penicillium chrysogenum

2017 ◽  
Vol 84 (4) ◽  
Author(s):  
Annarita Viggiano ◽  
Oleksandr Salo ◽  
Hazrat Ali ◽  
Wiktor Szymanski ◽  
Peter P. Lankhorst ◽  
...  
Keyword(s):  
Gene Cluster ◽  
Filamentous Fungi ◽  
Penicillium Chrysogenum ◽  
Biosynthetic Pathway ◽  
Biosynthetic Gene Cluster ◽  
Culture Broth ◽  
Yellow Pigment ◽  
Metabolic Potential ◽  
Content Type ◽  
Related Compounds

ABSTRACT Chrysogine is a yellow pigment produced by Penicillium chrysogenum and other filamentous fungi. Although the pigment was first isolated in 1973, its biosynthetic pathway has so far not been resolved. Here, we show that deletion of the highly expressed nonribosomal peptide synthetase (NRPS) gene Pc21g12630 ( chyA ) resulted in a decrease in the production of chrysogine and 13 related compounds in the culture broth of P. chrysogenum . Each of the genes of the chyA -containing gene cluster was individually deleted, and corresponding mutants were examined by metabolic profiling in order to elucidate their function. The data suggest that the NRPS ChyA mediates the condensation of anthranilic acid and alanine into the intermediate 2-(2-aminopropanamido)benzoic acid, which was verified by feeding experiments of a ΔchyA strain with the chemically synthesized product. The remainder of the pathway is highly branched, yielding at least 13 chrysogine-related compounds. IMPORTANCE Penicillium chrysogenum is used in industry for the production of β-lactams, but also produces several other secondary metabolites. The yellow pigment chrysogine is one of the most abundant metabolites in the culture broth, next to β-lactams. Here, we have characterized the biosynthetic gene cluster involved in chrysogine production and elucidated a complex and highly branched biosynthetic pathway, assigning each of the chrysogine cluster genes to biosynthetic steps and metabolic intermediates. The work further unlocks the metabolic potential of filamentous fungi and the complexity of secondary metabolite pathways.

scholarly journals Heme Protein and Hydroxyarginase Necessary for Biosynthesis of d-Cycloserine

2012 ◽  
Vol 56 (7) ◽  
pp. 3682-3689 ◽  
Author(s):  
Takanori Kumagai ◽  
Kisho Takagi ◽  
Yusuke Koyama ◽  
Yasuyuki Matoba ◽  
Kosuke Oda ◽  
...  
Keyword(s):  
Biosynthetic Pathway ◽  
Culture Medium ◽  
Biosynthetic Gene Cluster ◽  
Heme Protein ◽  
Content Type ◽  
Streptomyces Lavendulae ◽  
Spectroscopic Analyses ◽  
Encoding Gene ◽  
Oxide Synthase ◽  
Hydrolysis Of

ABSTRACTWe have recently cloned ad-cycloserine (DCS) biosynthetic gene cluster that consists of 10 genes, designateddcsA∼dcsJ, fromStreptomyces lavendulaeATCC 11924 (16). In the predicted pathway of hydroxyurea (HU) formation in DCS biosynthesis,l-arginine (L-Arg) must first be hydroxylated, prior to the hydrolysis ofNω-hydroxy-l-arginine (NHA) by DcsB, an arginase homolog. The hydroxylation of L-Arg is known to be catalyzed by nitric oxide synthase (NOS). In this study, to verify the supply route of HU, we created adcsB-disrupted mutant, ΔdcsB. While the mutant lost DCS productivity, its productivity was restored by complementation ofdcsB, and also by the addition of HU but not NHA, suggesting that HU is supplied by DcsB. A NOS-encoding gene,nos, fromS. lavendulaechromosome was cloned, to create anos-disrupted mutant. However, the mutant maintained the DCS productivity, suggesting that NOS is not necessary for DCS biosynthesis. To clarify the identity of an enzyme necessary for NHA formation, adcsA-disrupted mutant, designated ΔdcsA, was also created. The mutant lost DCS productivity, whereas the DCS productivity was restored by complementation ofdcsA. The addition of NHA to the culture medium of ΔdcsAmutant was also effective to restore DCS production. These results indicate that thedcsAgene product, DcsA, is an enzyme essential to generate NHA as a precursor in the DCS biosynthetic pathway. Spectroscopic analyses of the recombinant DcsA revealed that it is a heme protein, supporting an idea that DcsA is an enzyme catalyzing hydroxylation.

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

2012 ◽  
Vol 78 (14) ◽  
pp. 4908-4913 ◽  
Author(s):  
Bjarne Gram Hansen ◽  
Ewelina Mnich ◽  
Kristian Fog Nielsen ◽  
Jakob Blæsbjerg Nielsen ◽  
Morten Thrane Nielsen ◽  
...  
Keyword(s):  
Cytochrome P450 ◽  
Gene Cluster ◽  
Mycophenolic Acid ◽  
Polyketide Synthase ◽  
Fusion Gene ◽  
Bioinformatic Analysis ◽  
Terminal Region ◽  
Ring Closure ◽  
Content Type ◽  
Penicillium Brevicompactum

ABSTRACTMycophenolic 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 inPenicillium 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 inP. brevicompactumby obtaining full-length sequence of thempaDEcDNA prepared from the extracted RNA. Heterologous coexpression ofmpaCand the fusion genempaDEin the MPA-nonproducerAspergillus nidulansresulted in the production of 5,7-dihydroxy-4-methylphthalide (DHMP), the second intermediate in MPA biosynthesis. Analysis of the strain coexpressingmpaCand thempaDpart ofmpaDEshows 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.

scholarly journals Moorea producens gen. nov., sp. nov. and Moorea bouillonii comb. nov., tropical marine cyanobacteria rich in bioactive secondary metabolites

2012 ◽  
Vol 62 (Pt_5) ◽  
pp. 1171-1178 ◽  
Author(s):  
Niclas Engene ◽  
Erin C. Rottacker ◽  
Jan Kaštovský ◽  
Tara Byrum ◽  
Hyukjae Choi ◽  
...  
Keyword(s):  
Secondary Metabolites ◽  
Polyketide Synthase ◽  
Botanical Nomenclature ◽  
Distinctive Character ◽  
Content Type ◽  
Algae Blooms ◽  
Rich Diversity ◽  
Peptide Synthetase ◽  
Bioactive Secondary Metabolites ◽  
Micro Organisms

The filamentous cyanobacterial genus Moorea gen. nov., described here under the provisions of the International Code of Botanical Nomenclature, is a cosmopolitan pan-tropical group abundant in the marine benthos. Members of the genus Moorea are photosynthetic (containing phycocyanin, phycoerythrin, allophycocyanin and chlorophyll a), but non-diazotrophic (lack heterocysts and nitrogenase reductase genes). The cells (discoid and 25–80 µm wide) are arranged in long filaments (<10 cm in length) and often form extensive mats or blooms in shallow water. The cells are surrounded by thick polysaccharide sheaths covered by a rich diversity of heterotrophic micro-organisms. A distinctive character of this genus is its extraordinarily rich production of bioactive secondary metabolites. This is matched by genomes rich in polyketide synthase and non-ribosomal peptide synthetase biosynthetic genes which are dedicated to secondary metabolism. The encoded natural products are sometimes responsible for harmful algae blooms and, due to morphological resemblance to the genus Lyngbya , this group has often been incorrectly cited in the literature. We here describe two species of the genus Moorea: Moorea producens sp. nov. (type species of the genus) with 3LT as the nomenclature type, and Moorea bouillonii comb. nov. with PNG5-198R as the nomenclature type.

scholarly journals Aspergillus fumigatus SidJ Mediates Intracellular Siderophore Hydrolysis

2013 ◽  
Vol 79 (23) ◽  
pp. 7534-7536 ◽  
Author(s):  
Mario Gründlinger ◽  
Fabio Gsaller ◽  
Markus Schrettl ◽  
Herbert Lindner ◽  
Hubertus Haas
Keyword(s):  
Aspergillus Fumigatus ◽  
Gene Cluster ◽  
Biosynthetic Gene Cluster ◽  
Protein Family ◽  
Biosynthetic Gene ◽  
Intracellular Accumulation ◽  
Iron Starvation ◽  
Content Type ◽  
Hydrolysis Products ◽  
Encoding Gene

ABSTRACTSiderophore-mediated iron handling is crucial for the virulence ofAspergillus fumigatus. Here we identified a new component of its siderophore metabolism, termed SidJ, which is encoded by AFUA_3G03390. The encoding gene is localized in a siderophore biosynthetic gene cluster that is conserved in a variety of fungi. During iron starvation, SidJ deficiency resulted in decreased growth and increased intracellular accumulation of hydrolysis products of the siderophore fusarinine C. The implied role in siderophore hydrolysis is consistent with a putative esterase domain in SidJ, which now represents the first functionally characterized member of the DUF1749 (domain ofunknownfunction) protein family, with members found exclusively in fungi and plants.

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