Fungus Phoma Sp Research Articles

Fungal polyketides produced by an endophytic fungus Phoma sp. associated with Gastrodia elata

Two novel fungal polyketides, phometides A (1) and B (2), together with four known compounds (3–6), were isolated from the endophytic fungus Phoma sp. YUD17001 obtained from Gastrodia elata Blume. The structures were elucidated based on spectroscopic analyses, X-ray crystal diffraction, and time-dependent density functional theory/electronic circular dichroism (TDDFT/ECD) calculations. Structurally, phometide A (1) represented the first example of C12 polyketide characterized by an unusual tetrahydrobenzofuran-3(2H)-one core with an α,β-unsaturated ketone functionality, while phometide B (2) was an unprecedented molecule containing a 2-pentylcycloheptan-1-one scaffold. In an antimicrobial activity assay, phometide A (1) exhibited significant inhibitory activity against Staphylococcus aureus with MIC value of 4 μg/mL. Phometide B (2) showed moderate antifungal activity against Candida albicans with an MIC value of 16 μg/mL. Furthermore, compounds 1 and 2 were evaluated for their acetylcholinesterase inhibitory and cytotoxic activities.

Pyranone Derivatives With Antitumor Activities, From the Endophytic Fungus Phoma sp. YN02-P-3

Two new pyranone derivatives phomapyrone A (2) and phomapyrone B (3), one new coumarin 11S, 13R-(+)-phomacumarin A (1), three known pyranones (4–6), together with three known amide alkaloids fuscoatramides A-C (7–9), as well as 9S, 11R-(+)-ascosalitoxin (10) were isolated from the endophytic fungus Phoma sp. YN02-P-3, which was isolated from the healthy leaf tissue of a Paulownia tree in Yunnan Province, China. Their structures were elucidated using extensive NMR spectroscopic and HRESIMS data and by comparing the information with literature data. In addition, all compounds were tested for their cytotoxicity activity against human tumor cell lines, and the results showed that new compounds 1-3 showed moderate inhibitory activity against the HL-60 cell line with IC50 values of 31.02, 34.62, and 27.90 μM, respectively.

Characterization and biological activities of melanin pigment from root endophytic fungus, Phoma sp. RDSE17.

Melanins are high molecular weight hydrophobic pigments which have gained popularity for their role in virulence against different pathogens. In the present study, we isolated and characterized the melanin pigment produced by a dark septate endophyte fungus Phoma sp. RDSE17, which was associated with the roots of an indigenous Oryza sativa cv. 'Chakhao amubi' in Manipur, Northeast India. The biological properties of purified melanin from the fungus were evaluated for their antioxidant, antimicrobial and anticancerous activities. The pigment was extracted from Phoma sp. by alkaline-acid hydrolysis method and confirmed as melanin through physico-chemical tests and spectral (UV, FTIR, and EPR) analysis. The analyses of the elemental composition indicated that the pigment possessed a low percentage of nitrogen (N) contents, and therefore, would not fall under DOPA class of melanin. Exposure of the fungus to melanin pathway inhibitors revealed a positive melanin inhibition by tricyclazole, but not by kojic acid. Thus, the melanin from Phoma sp. may be a member of the DHN family. Moreover, the purified melanin showed high DPPH (1, 1-Diphenyl-2-picrylhydrazyl) free radical-scavenging activity with an EC50 of 69µg/mL and inhibited human lung cancer cell (A549 cells) proliferation at 80µg/mL. The present study demonstrates that melanin from Phoma sp. RDSE17 could be employed as a potential biological (antioxidant) and antimicrobial agent for inhibiting the growth of humans and phytopathogens.

A new anti-proliferative compound from an endophytic fungus, Phoma sp.

A new oxazole-type compound (1), named macrooxazole E, and three known macrooxazoles A-C (2–4), were isolated from ethyl acetate (EtOAc) extracts of Phoma sp. JS0228 cultures, an endophytic fungus of Morus alba (M. alba). Structures of the isolated compounds were determined using spectroscopic methods, such as 1 D- and 2 D-nuclear magnetic resonance (NMR) and high-resolution mass spectrometry (HRMS). Macrooxazole E (1) differed from macrooxazole C only in the presence of methyl carboxylate instead of free carboxylic acid. Macrooxazole C showed moderate anti-proliferative activities against human breast cancer (MCF-7) and prostate cancer (LNCaP) cell lines with IC50 values of 0.29 mM and 0.36 mM, respectively. This study presents the possibility of the endophytic fungus Phoma sp. JS0228 to produce new bioactive natural compounds.

A New Diphenyl Ether Derivative from a Mangrove Endophytic Fungus Phoma sp. L28

A New Diphenyl Ether Derivative from a Mangrove Endophytic Fungus Phoma sp. L28

Metabolites with anti-inflammatory and α-glucosidase inhibitory activities from the mangrove endophytic fungus Phoma sp. SYSU-SK-7

Four new steroids phomosterols A-B (1–2), phomopenes A-B (3–4) and one known analogue (5) were isolated from the mangrove endophytic fungus Phoma sp. SYSU-SK-7. Their structures and absolute configurations were determined via NMR data, single-crystal X-ray diffraction analysis, and ECD calculations. Compounds 1–2 contain an unusual aromatic B ring skeleton in sterols, and compound 1 represent the first crystal structure described. Compounds 1, 2 and 5 showed inhibitory activities against nitric oxide (NO) production in lipopolysaccharides (LPS) induced RAW 264.7 cells with IC50 values of 13.5, 25.0 and 12.5 μM, respectively. Then, the affinities between new bioactive compounds (1 and 2) with nitric oxide synthase (iNOS) were researched by molecular docking studies.

Phomretones A-F, C12 polyketides from the co-cultivation of Phoma sp. YUD17001 and Armillaria sp.

Six new C12 polyketides, phomretones A–F (1–6), were isolated from the co-culture of Armillaria sp. and the endophytic fungus Phoma sp. YUD17001 associated with Gastrodia elata. Neither fungus produced these compounds when cultured alone. The structures of 1–6 were established on the basis of comprehensive spectroscopic analyses, while their absolute configurations were determined by the comparsion of experimental and calculated ECD spectra. Compounds 2–4 are diastereoisomers of each other and featured high levels of stereoisomerization and oxidation.

Bioactive polyketides from the mangrove endophytic fungi Phoma sp. SYSU-SK-7

Five new polyketides, colletotric B (2), 3-hydroxy-5-methoxy-2,4,6-trimethylbenzoic acid (3), colletotric C (4), chaetochromone D (6) and 8-hydroxy-pregaliellalactone B (9), together with four known analogues (1, 5 and 7–8) were isolated from the mangrove endophytic fungus Phoma sp. SYSU-SK-7. Their structures were elucidated by analysis of extensive spectroscopic data and mass spectrometric data. Compounds 1–2 showed strong antimicrobial activity against the P. aeruginosa, MRSA and C. albicans with the MIC values in the range of 1.67–6.28 μg/ml. Furthermore, Compounds 1–5 also exhibited significant α-glucosidase inhibitory activity with the IC50 values in the range of 36.2–90.6 μM. Compound 7 was found to inhibited radical scavenging activity against DPPH with the EC50 value of 11.8 μM.

Antioxidant epoxydon and benzolactone derivatives from the insect-associated fungus Phoma sp.

One new epoxydon ester (1) and a new benzolactone derivative (2), along with four known compounds (3−6), were isolated from the insect-associated fungus Phoma sp. Their structures were confirmed by extensive MS and NMR spectroscopic analysis and their absolute configurations were determined by a combination of modified Mosher method and Mo2(OCOCH3)4-induced electronic circular dichroism (ECD) experiments. Compounds 1 and 5 were revealed to have potent antioxidant activities, which were approximate to the potency of the positive control trolox. In addition, 1 also exhibited moderate cytotoxic effect against human MGC-803 tumor cell line.

Genomics-driven discovery of a biosynthetic gene cluster required for the synthesis of BII-Rafflesfungin from the fungus Phoma sp. F3723

BackgroundPhomafungin is a recently reported broad spectrum antifungal compound but its biosynthetic pathway is unknown. We combed publicly available Phoma genomes but failed to find any putative biosynthetic gene cluster that could account for its biosynthesis.ResultsTherefore, we sequenced the genome of one of our Phoma strains (F3723) previously identified as having antifungal activity in a high-throughput screen. We found a biosynthetic gene cluster that was predicted to synthesize a cyclic lipodepsipeptide that differs in the amino acid composition compared to Phomafungin. Antifungal activity guided isolation yielded a new compound, BII-Rafflesfungin, the structure of which was determined.ConclusionsWe describe the NRPS-t1PKS cluster ‘BIIRfg’ compatible with the synthesis of the cyclic lipodepsipeptide BII-Rafflesfungin [HMHDA-L-Ala-L-Glu-L-Asn-L-Ser-L-Ser-D-Ser-D-allo-Thr-Gly]. We report new Stachelhaus codes for Ala, Glu, Asn, Ser, Thr, and Gly. We propose a mechanism for BII-Rafflesfungin biosynthesis, which involves the formation of the lipid part by BIIRfg_PKS followed by activation and transfer of the lipid chain by a predicted AMP-ligase on to the first PCP domain of the BIIRfg_NRPS gene.

Inducing Secondary Metabolite Production by Co-culture of the Endophytic Fungus Phoma sp. and the Symbiotic Fungus Armillaria sp.

Co-culturing the endophytic fungus Phoma sp. YUD17001 from Gastrodia elata with Armillaria sp. in liquid nutrient medium resulted in the production of five new secondary metabolites, including two phenolic compounds, phexandiols A and B (1 and 2), three aliphatic ester derivatives, phomesters A-C (3-5), and a known fatty acid (6). The structures and absolute configurations of these compounds were elucidated by the interpretation of data from detailed spectroscopic analysis, Mosher's method, and electronic circular dichroism spectra, together with consideration of the biogenetic origins. None of the five new compounds were detected in single-strain cultures under identical fermentation conditions. The results of this work indicated that the production of 1-5 involved a complicated interaction process. None of the new compounds possessed significant cytotoxicity or antimicrobial activities.

Bioactive secondary metabolites from an endophytic fungus Phoma sp. PF2 derived from Artemisia princeps Pamp.

Two new isochromanone derivatives, (3S,4S)-3,8-dihydroxy-6-methoxy-3,4,5-trimethylisochroman-1-one (1) and methyl (S)-8-hydroxy-6-methoxy-5-methyl-4a-(3-oxobutan-2-yl)benzoate (2), together with six known compounds (3‒8) were isolated from thecultures of an endophytic fungus Phoma sp. PF2 obtained from Artemisia princeps. The chemical structures of the isolated compounds were elucidated by interpretation of spectroscopic data (1D, 2D NMR, HRESIMS, and CD) and calculation of ECD. All the isolated compounds (1‒8) showed moderate inhibitory activities on nitric oxide levels in lipopolysaccharide-induced RAW264.7 machrophage cells.

New bioactive chlorinated cyclopentene derivatives from the marine-derived Fungus Phoma sp

The new halogenated metabolites cryptophomic acid (1), cryptodiol (2), and cryptotriol (3) and the known dihydro-isocoumarin derivative 4, were isolated from the marine-derived fungus Phoma sp.135. The structural elucidation of these compounds was achieved by extensive analysis of spectroscopic data including 1D- and 2D-NMR, HRMS, optical rotation, UV, and IR. The absolute configuration of cryptophomic acid (1) was determined by using circular dichroism and TD-DFT ECD calculations of the solution conformers. The relative configurations of cryptodiol (2) and cryptotriol (3) were elucidated by a detailed analysis of the spectroscopic data and quantum mechanical calculation of NMR chemical shifts, based also on the newly reported + approach. The dihydro-isocoumarin derivative 4 was also produced by the same fungus and its structure was established as (R)-8-hydroxy-6-methoxy-3-methy-3,4-dihydro-isocoumarin (4), often referred to 6-methoxymellein and its chemical structure and absolute configuration were further confirmed through X-ray diffraction analysis. We report here the new compounds 1–3 and the crystallographic data of compound 4 for the first time. Additionally, we report significant antimicrobial activity of compounds 1–3 against Escherichia coli, Bacillus subtilis, Mycobacterium phlei, and Staphylococcus aureus and they showed no lethality against brine shrimp.

Anti-inflammatory phomalichenones from an endolichenic fungus Phoma sp.

Four new compounds, phomalichenones A-D (1-4), and seven known compounds (5-11) were isolated from the cultures of an endolichenic fungus Phoma sp. EL002650. Their structures were determined by the analysis of their spectroscopic data (NMR and MS). Compounds 1 and 6 inhibited nitric oxide (NO) production in lipopolysaccharide (LPS)-stimulated RAW264.7 macrophages. In addition, compound 1 diminished the protein expression levels of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2), and decreased the mRNA expression levels of pro-inflammatory cytokines, such as tumor necrosis factor-α (TNF-α), interleukin(IL)-1β, and IL-6.

α-Pyrone derivatives with cytotoxic activities, from the endophytic fungus Phoma sp. YN02-P-3

Four new α-pyrone derivatives phomones C-F (1−4) together with four known compounds (5−8) were isolated from the endophytic fungus Phoma sp. YN02-P-3. Compound 1 is the first example of 6-α,β-unsaturated ester-2-pyrone dimers via intermolecular symmetrical [2+2] cycloaddition. The chemical structures of these compounds were determined from spectroscopic data (1D/2D NMR, MS and IR). The acetylated product (9) of 1 along with compounds 1–8 were then tested for their cytotoxicity against HL-60, PC-3 and HCT-116 cell lines. Compounds 2, 3, 5 and 9 with acetyl groups showed significant inhibitory activities against the three cell lines with IC50 values in the range 0.52–9.85μM. while compounds 1, 4 and 6–8 that possess no acetyl group showed no inhibitory activity (IC50>50μM), indicating that the acetyl group at 10- or 12- are essential for their cytotoxic activities. The structure-activity relationships of these phomones were also reported.

Identification and Antifungal Activity of Metabolites from the Mangrove Fungus Phoma sp. L28

A new anthraquinone, 7-(γ,γ)-dimethylallyloxymacrosporin (1), along with five known analogues, macrosporin (2), 7-methoxymacrosporin (3), tetrahydroaltersolanol B (4), altersolanol L (5), and ampelanol (6), were isolated from the mangrove endophytic fungus Phoma sp. L28. All of them were first found in Phoma sp. Their structures were established by comprehensive spectroscopic analyses and comparison with the published data. These compounds displayed in vitro antifungal activities against Colletotrichum musae (Berk. & M. A. Curtis) Arx., Colletotrichum gloeosporioides (Penz) Sacc., Fusarium graminearum Schw., Penicillium italicum Wehme, Fusarium oxysporum Schlecht. f. sp. lycopersici (Sacc.) W.C. Snyder et H. N. Hansen, and Rhizoctonia solani Kuhn at different levels. Notably, compound 2 exhibited potent antifungal activity against Fusarium graminearum Schw. as compared with the positive control carbendazim.

Biphenyl ether derivatives with protein tyrosine phosphatase 1B inhibitory activity from the freshwater fungus Phoma sp.

Biphenyl ether derivatives with protein tyrosine phosphatase 1B inhibitory activity from the freshwater fungus Phoma sp.

Two pairs of enantiomeric α-pyrone dimers from the endophytic fungus Phoma sp. YN02-P-3

(±) Phomones A (1) and B (2), two pairs of novel enantiomeric α-pyrone dimers from the endophytic fungus Phoma sp. YN02-P-3 are reported.

A novel 3,4-dihydronaphthalen-1(2H)-one with spiro-butyrolactone and a new isocoumarin isolated from the endophytic fungus Phoma sp. YN02-P-3

A novel 3,4-dihydronaphthalen-1(2H)-one with spiro-butyrolactone phomol (1) and a new isocoumarin phomasatin (2), together with two known compounds (3–4) were isolated from the solid culture of the endophytic fungus Phoma sp. YN02-P-3. Their structures including the absolute configurations were characterized on the basis of extensive 1D, 2D NMR (HSQC, HMBC, NOESY), MS, and CD spectral data. Compound 1 showed selective cytotoxic activity against HL-60 cell line with the IC50 value of 29.05 μM.

Identification and Heterologous Expression of the Topopyrone Nonaketide Synthase Gene from Phoma sp.

Non-reducing iterative type I polyketide synthase genes, pnk1 and pnk2, were cloned from the fungus Phoma sp. BAUA2861, which produces the topoisomerase I inhibitors, topopyrones A to D. Heterologous expression of these polyketide synthase genes under the α-amylase promoter in Aspergillus oryzae was carried out to identify their functions. The pnk2 transformant produced topopyrones C, D, and haematommone. Therefore, the pnk2 gene was found to encode for the topopyrone nonaketide synthase.