Ali Wang

Bis-naphtho-γ-pyrones from Fungi and Their Bioactivities

Bis-naphtho-γ-pyrones are an important group of aromatic polyketides derived from fungi. They have a variety of biological activities including cytotoxic, antitumor, antimicrobial, tyrosine kinase and HIV-1 integrase inhibition properties, demonstrating their potential applications in medicine and agriculture. At least 59 bis-naphtho-γ-pyrones from fungi have been reported in the past few decades. This mini-review aims to briefly summarize their occurrence, biosynthesis, and structure, as well as their biological activities. Some considerations regarding to synthesis, production, and medicinal and agricultural applications of bis-naphtho-γ-pyrones are also discussed.

Bioactive Bis-naphtho-γ-pyrones from Rice False Smut Pathogen Ustilaginoidea virens

Ustilaginoidins were bis-naphtho-γ-pyrones mycotoxins possessing an aR configuration of the chiral axis previously reported from the false smut balls of rice infected by the fungal pathogen Ustilaginoidea virens. To investigate the chemical diversity of these metabolites and their bioactivities, we fermented this fungus on solid rice media, which afforded the isolation of 13 ustilaginoidins, including seven new compounds, namely ustilaginoidins K-P, 1-6, and E1, 7, together with the known ustilaginoidins A, 8, D, 9, E, 10, F, 11, and G, 12, and isochaetochromin B2, 13. The structures of the new compounds were elucidated by using (1D, 2D) NMR, high-resolution mass spectrometry, UV, and circular dichroism, as well as by comparison with the literature data. A plausible biosynthesis pathway was proposed for these dimeric polyketides. The isolated compounds were evaluated for their antibacterial, cytotoxic, and radicle elongation inhibitory activities. Ustilaginoidins K, 1 and L, 2 showed cytotoxic activities on the A2780 human ovarian cancer cell line with IC50 values of 4.18 and 7.26 μM, respectively. Ustilaginoidins N, 4, D, 9, E, 10, and G, 12 were active against the tested pathogenic bacteria with MIC values in the range of 16-64 μg/mL. Ustilaginoidins O, 5, E, 10, and F, 11, and isochaetochromin B2, 13 displayed moderate inhibitory activity on the radicle elongation of rice seeds.

Development of a Monoclonal Antibody-Based icELISA for the Detection of Ustiloxin B in Rice False Smut Balls and Rice Grains

Rice false smut is an emerging and economically-important rice disease caused by infection by the fungal pathogen Villosiclava virens. Ustiloxin B is an antimitotic cyclopeptide mycotoxin isolated from the rice false smut balls that formed in the pathogen-infected rice spikelets. A monoclonal antibody (mAb) designated as mAb 1B5A10 was generated with ustiloxin B—ovalbumin conjugate. A highly-sensitive and specific indirect competitive enzyme-linked immunosorbent assay (icELISA) was then developed. The median inhibitory concentration (IC50) of the icELISA was 18.0 ng/mL for the detection of ustiloxin B; the limit of detection was 0.6 ng/mL, and the calibration range was from 2.5 to 107.4 ng/mL. The LOD/LOQ values of the developed ELISA used for the determination of ustiloxin B in rice false smut balls and rice grains were 12/50 μg/g and 30/125 ng/g, respectively. The mAb 1B5A10 cross-reacted with ustiloxin A at 13.9% relative to ustiloxin B. Average recoveries of ustiloxin B ranged from 91.3% to 105.1% for rice false smut balls at spiking levels of 0.2 to 3.2 mg/g and from 92.6% to 103.5% for rice grains at spiking levels of 100 to 5000 ng/g. Comparison of ustiloxin B content in rice false smut balls and rice grains detected by both icELISA and high performance liquid chromatography (HPLC) demonstrated that the developed icELISA can be employed as an effective and accurate method for the detection of ustiloxin B in rice false smut balls, as well as rice food and feed samples.

Bioactive Dibenzo-α-pyrone Derivatives from the Endophytic Fungus Rhizopycnis vagum Nitaf22

Six new dibenzo-α-pyrones, rhizopycnolides A (1) and B (2) and rhizopycnins A-D (3-6), together with eight known congeners (7-14), were isolated from the endophytic fungus Rhizopycnis vagum Nitaf22 obtained from Nicotiana tabacum. The structures of the new compounds were unambiguously elucidated using NMR, HRESIMS, TDDFT ECD calculation, and X-ray crystallography data. Rhizopycnolides A (1) and B (2) feature an uncommon γ-butyrolactone-fused dibenzo-α-pyrone tetracyclic skeleton (6/6/6/5), while rhizopycnin B (4) was the first amino group containing dibenzo-α-pyrone. Rhizopycnolides A (1) and B (2) are proposed to be biosynthesized from polyketide and tricarboxylic acid cycle pathways. The isolated compounds were tested for their antibacterial, antifungal, and cytotoxic activities. Among them, rhizopycnolide A (1), rhizopycnins C (5) and D (6), TMC-264 (8), penicilliumolide D (11), and alternariol (12) were active against the tested pathogenic bacteria Agrobacterium tumefaciens, Bacillus subtilis, Pseudomonas lachrymans, Ralstonia solanacearum, Staphylococcus hemolyticus, and Xanthomonas vesicatoria with MIC values in the range 25-100 μg/mL. Rhizopycnin D (6) and TMC-264 (8) strongly inhibited the spore germination of Magnaporthe oryzae with IC50 values of 9.9 and 12.0 μg/mL, respectively. TMC-264 (8) showed potent cytotoxicity against five human cancer cell lines (HCT-116, HepG2, BGC-823, NCI-H1650, and A2780) with IC50 values of 3.2-7.8 μM.

A monoclonal antibody-based enzyme-linked immunosorbent assay for detection of ustiloxin A in rice false smut balls and rice samples.

Ustiloxin A, a cyclopeptide mycotoxin, was isolated from the pathogenic fungus Villosiclava virens that causes rice false smut, a worldwide devastating rice disease. A monoclonal antibody (mAb) 2D3G5 was generated with ustiloxin A-bovine serum albumin conjugate. A highly sensitive and specific indirect competitive enzyme-linked immunosorbent assay (icELISA) was then developed. It possessed a median inhibition concentration (IC50) of 13.8 ng/mL and a working range of 2.8-72 ng/mL. The mAb 2D3G5 recognized ustiloxin B with the cross-reactivity as 4%. The average recoveries of ustiloxin A from rice false smut balls and peeled rice samples ranged from 92% to 117% and from 92% to 107%, respectively. Comparison of the concentrations of ustiloxin A in rice false smut balls detected by both icELISA and high performance liquid chromatography-photodiode array detection indicated that the developed icELISA was suitable for detection of ustiloxin A in rice food and feed samples.

The Contents of Ustiloxins A and B along with Their Distribution in Rice False Smut Balls

Ustiloxins are cyclopeptide mycotoxins isolated from rice false smut balls (FSBs), the ball-like colonies transformed from the individual grains through the filament infection by the fungal pathogen Villosiclava virens. There were no obvious relations between ustiloxin content and any of the collection areas, collection times, or average weight of each FSB. The rice false smut balls at early, middle, and late maturity stages were respectively divided into different parts (glume, chlamydospores, mycelia, and pseudoparenchyma). The highest content of ustiloxins A and B of rice FSBs was found at the early maturity stage. Both ustiloxins A and B were mainly distributed in the middle layer containing mycelia and immature chlamydospores of the FSBs. When the rice FSBs were at the early maturity stage, the total yield of ustiloxins A and B in the middle layer of each ball was 48.3 µg, which was 3.20-fold of the yield (15.1 µg) of the inner part of the ball. The rice FSBs at the early maturity stage are the appropriate materials for the production of ustiloxins A and B.

A new proline-containing flavonol glycoside from Caragana leucophloea Pojark

One new proline-containing flavonol glycoside, namely kaempferol-3-O-methyl-7-O-β-d-glucopyranosyl-8-(1-methyleneproline)-4′-O-β-d-glucopyranoside (1), together with 15 known flavonoids, 3-O-methylkaempferol (2), 3-O-methylquercetin (3), quercetin (4), kaempferol (5), apigenin (6), rhamnazin (7), astragalin (8), alquds (9), quercitrin (10), rutin (11), isoquercitrin (12), apigetrin (13), myricitrin (14), hesperidin (15) and calycosin-7-O-β-d-glucopyranoside (16) were isolated from the aerial parts of Caragana leucophloea Pojark. (Leguminosae). Their structures were determined on the basis of spectroscopic analyses and by comparison with literature data. Compounds 2–4 revealed a strong antimicrobial activity with minimum inhibitory concentration values of 12.5–150 μg/mL and median inhibitory concentration (IC50) values of 7.42–76.61 μg/mL. Compounds 3, 4, 6–8, 10–12 and 14 showed strong antioxidant activity. Compounds 2–7 exhibited moderate antinematodal activity on Caenorhabditis elegans with IC50 values of 40.51–68.05 μg/mL.

New Ustilaginoidins from Rice False Smut Balls Caused by Villosiclava virens and Their Phytotoxic and Cytotoxic Activities

Ustilaginoidins are a class of bis-naphtho-γ-pyrones, typically produced by Villosiclava virens, the pathogen of the rice false smut (RFS), which has been one of the most destructive rice fungal diseases. Previously, we found that ustilaginoidins identified from the culture of V. virens on rice medium were less polar than those reported from the RFS balls in general. In this study, we reinvestigated the high-performance liquid chromatography with diode array detection and high-resolution mass spectrometry (HPLC-DAD-HRMS) profile of the ethyl acetate (EtOAc) extract of the RFS balls and found several interesting peaks that correspond to new ustilaginoidins. As a result, eight new and polar congeners, named ustilaginoidins Q-T (1-4), 2,3-dihydroustilaginoidin T (5), and ustilaginoidins U-W (6-8), were isolated. In addition, 17 known ustilaginoidins, including ustilaginoidins K-N (9-12), ustilaginoidin P (13), ustilaginoidin E1 (14), isochaetochromin B2 (15), and ustilaginoidins A-J (16-25), were re-isolated. The structures of the new compounds were elucidated by comprehensive analysis of the spectroscopic data. Ustilaginoidins Q (1) and R (2) feature an uncommon 2-hydroxypropyl-substituted skeleton and biogenetically incorporate one more acetate unit than common ustilaginoidins. Ustilaginoidin W (8) is a rare formate-containing bis-naphtho-γ-pyrone. Ustilaginoidins R (2), U (6), B (17), and I (24) showed moderate inhibitory activities toward the radicle or germ elongation of rice seeds. Ustilaginoidins R (2), S (3), V (7), W (8), B (17), C (18), and H-J (23-25) were cytotoxic to the tested human cancer cell lines (HCT116, NCI-H1650, BGC823, Daoy, and HepG2), with IC50 values in the range of 4.06-44.1 μM.

Hyalodendriellins A–F, new 14-membered resorcylic acid lactones from the endophytic fungus Hyalodendriella sp. Ponipodef12

Six new 14-membered resorcylic acid lactones (RALs), named hyalodendriellins A–F (1–6), were isolated from the culture of the endophytic fungus Hyalodendriella sp. Ponipodef12 associated with the hybrid ‘Neva’ of Populus deltoides Marsh × P. nigra L. The structures of the new compounds were deduced by analyses of the NMR spectroscopic and mass spectrometry data, in combination with chemical conversion, modified Moshers method and TDDFT ECD calculations for determining the absolute configurations. Compounds 1, 2, 4, and 6 possess a 3S configuration, while 3 and 5 are 3R-configurated. The co-occurrence of RALs with different stereochemistry at C-3 in the same fungus is rare. The CD behaviors of 1–6 as well as their acetonide and hydrogenated derivatives were investigated. All the isolated compounds were evaluated for their antinematodal, larvicidal, cytotoxic, antibacterial and antifungal activities. Among them, hyalodendriellin A (1) displayed moderate antinematodal activity against Caenorhabditis elegans and Meloidogyne incognita. Hyalodendriellin C (3) exhibited larvicidal effect against the fourth-instar larvae of the mosquito Aedes aegypti.

Enhancement of Palmarumycin C12 and C13 Production by the Endophytic Fungus Berkleasmium sp. Dzf12 in an Aqueous-Organic Solvent System

The endophytic fungus Berkleasmium sp. Dzf12, isolated from Dioscorea zingiberensis, was found to produce palmarumycins C12 and C13 which possess a great variety of biological activities. Seven biocompatible water-immiscible organic solvents including n-dodecane, n-hexadecane, 1-hexadecene, liquid paraffin, dibutyl phthalate, butyl oleate and oleic acid were evaluated to improve palmarumycins C12 and C13 production in suspension culture of Berkleasmium sp. Dzf12. Among the chosen solvents both butyl oleate and liquid paraffin were the most effective to improve palmarumycins C12 and C13 production. The addition of dibutyl phthalate, butyl oleate and oleic acid to the cultures of Berkleasmium sp. Dzf12 significantly enhanced palmarumycin C12 production by adsorbing palmarumycin C12 into the organic phase. When butyl oleate was fed at 5% (v/v) in medium at the beginning of fermentation (day 0), the highest palmarumycin C12 yield (191.6 mg/L) was achieved, about a 34.87-fold increase in comparison with the control (5.3 mg/L). n-Dodecane, 1-hexadecene and liquid paraffin had a great influence on the production of palmarumycin C13. When liquid paraffin was added at 10% (v/v) in medium on day 3 of fermentation, the palmarumycin C13 yield reached a maximum value (134.1 mg/L), which was 4.35-fold that of the control (30.8 mg/L). Application of the aqueous-organic solvent system should be a simple and efficient process strategy for enhancing palmarumycin C12 and C13 production in liquid cultures of the endophytic fungus Berkleasmium sp. Dzf12.