Synthesis and antifungal activity of novel sclerotiorin analogues

Sclerotiorin 1, first isolated from Penicillium sclerotiorum, has weak antifungal activity and belongs to the azaphilone-type family of natural products. Several series of sclerotiorin analogues were designed and synthesized with the aim of discovering novel fungicides with improved activity. The syntheses involved two key steps, cycloisomerization and then oxidation, and used a simple and efficient Sonogashira cross-coupling reaction to construct the required functionalized precursor. With sclerotiorin as a control, the activities of the newly synthesized analogues were evaluated against seven fungal pathogens, and several promising candidates (compounds 3a?, 3d?, 3e?, 3f? and 3k?) with greater activity and simpler structures than sclerotiorin were discovered. In addition, preliminary structure-activity relationships were studied, which revealed that not only the chlorine or bromine substituent at the 5-position of the nucleus but also the phenyl group at the 3-position and the substituent pattern on it contributed crucially to the observed antifungal activity. Analogues with a methyl substituent at the 1-position have reduced levels of activity, while those with a free hydroxyl group in place of acetoxy at the quaternary center of the bicyclic ring system retain activity.     

Isoflavones as potentiators of antibacterial activity

Isoflavones isolated from Lupinus argenteus were found to potentiate the antibacterial activity of alpha-linolenic acid, also found in the same plant. The isoflavones also potentiated the activity of the natural plant antibiotic berberine and the synthetic fluoroquinoline antibiotic norfloxacin. The isoflavones increased the uptake of berberine into Staphylococcus aureus cells, indicating that they may be inhibiting a multidrug resistance pump (MDR). Thus, L. argenteus contains a weak antibacterial and also MDR pump inhibitors, which increase its potency.     

pH modulation of zopfiellin antifungal activity to Colletotrichum and Botrytis

Zopfiellin, a novel cyclooctanoid natural product isolated from Zopfiella curvata No. 37-3, was evaluated in a 96-well microtiter assay for fungicidal activity against Botrytis cinerea, Colletotrichum acutatum, Colletotrichum fragariae, Colletotrichum gloeosporioides, and Fusarium oxysporum. Zopfiellin exhibited pH-dependent activity, with the most mycelial growth inhibition demonstrated at pH 5.0. Mass spectrometry and nuclear magnetic resonance spectroscopy studies indicated that zopfiellin undergoes structural changes with changes in pH. At pH 5.0, zopfiellin showed the greatest activity against B. cinerea (IC(80) = 10 microM), C. gloeosporioides (IC(80) = 10 microM), and C. fragariae (IC(80) = 10 microM) and intermediate activity against C. acutatum (IC(80) = 30 microM), and was not active against F. oxysporum (IC(80) > 100 microM).     

Chemical composition and antifungal activity of essential oil of Chrysactinia mexicana gray

The chemical composition of the essential oil of Chysactinia mexicana was analyzed by gas chromatography-mass spectrometry. Seventeen compounds were characterized; eucalyptol (41.3%), piperitone (37.7%), and linalyl acetate (9.1%) were found as the major components. The essential oil of leaves and piperitone completely inhibited Aspergillus flavus growth at relatively low concentrations (1.25 and 0.6 mg/mL, respectively).     

Protein from red cabbage (Brassica oleracea) seeds with antifungal, antibacterial, and anticancer activities

A 30 kDa antifungal protein was purified from red cabbage ( Brassica oleracea ) seeds. It exhibited a molecular mass and N-terminal amino acid sequence disinct from those of previously isolated Brassica antifungal proteins. The protocol used entailed ion exchange chromatography on Q-Sepharose and SP-Sepharose followed by fast protein liquid chromatography on Mono S. The protein hindered mycelial growth in Mycosphaerella arachidicola (with an IC50=5 μM), Setospaeria turcica, and Bipolaris maydis. It also inhibited the yeast Candida albicans with an IC50=96 μM. It exerted its antifungal action by permeabilizing the fungal membrane as evidenced by staining with Sytox green. The antifungal activity was stable from pH 3 to 11 and from 0 to 65 °C. It manifested antibacterial activity against Pseudomonas aeruginosa (IC50=53 μM). Furthermore, after 48 h of culture, it suppressed proliferation of nasopharyngeal cancer and hepatoma cells with IC50=50 and 90 μM, respectively.     

西藏"灵菇"乳的发酵与抑菌机制研究

根据发酵过程中菌粒中主要微生物数量与发酵乳中主要营养物和产物含量间的关系,分析探讨了西藏"灵菇"乳的发酵机制:考察了不同发酵时期所得发酵乳对人肠杆菌、金黄色葡萄球菌、沙门氏菌的抑菌活性,结合化学物质模拟试验揭示了发酵乳的抑菌机制.结果显示,发酵前期,菌粒中的各微生物由于向乳中扩散及生长延滞,数量稍有减少;随着发酵的进行,菌粒中乳酸菌和酵母菌的数量一直处于相对稳定的水平,但醋酸菌含量呈明显的先升后降趋势;在整个发酵过程中,菌粒中的乳酸菌数量一直处于最高水平,其次为酵母菌,醋醋菌数量最少:乳酸是西藏"灵菇"发酵乳中的主要抑菌活性物质;乙酸和乙醇的存在对低浓度乳酸的抑菌活性有增效作用,但当乳酸浓度较高时,这种增效作用减弱.西藏"灵菇"发酵乳的总抑菌效果是微生物与其代谢产物的共同作用结果.     

Chemical composition and antifungal activity of Salvia macrochlamys and Salvia recognita essential oils

Essential oils of Salvia macrochlamys and Salvia recognita were obtained by hydrodistillation of dried aerial parts and characterized by gas chromatography and gas chromatography-mass spectrometry. One hundred and twenty identified constituents representing 97.7% in S. macrochlamys and 96.4% in S. recognita were characterized, and 1,8-cineole, borneol, and camphor were identified as major components of the essential oils. The oils were evaluated for their antimalarial, antimicrobial, and antifungal activities. Antifungal activity of the essential oils from both Salvia species was nonselective at inhibiting growth and development of reproductive stroma of the plant pathogens Colletotrichum acutatum, Colletotrichum fragariae, and Colletotrichum gloeosporioides. S. macrochlamys oil had good antimycobacterial activity against Mycobacterium intracellulare; however, the oils showed no antimicrobial activity against human pathogenic bacteria or fungi up to a concentration of 200 microg/mL. S. recognita oil exhibited a weak antimalarial activity against Plasmodium falciparum.     

Modeling production of antifungal compounds and their role in biocontrol product inhibitory activity

Partial least squares (PLS) regression modeling was used to relate the antifungal activity of Bacillus subtilis solid-state fermentation extracts to the individual high-performance liquid chromatography (HPLC) peaks from those extracts. A model was developed that predicted bioassay inhibition based on the extract HPLC profile (R(2) = 0.99). Concentrations of the members of the antifungal lipopeptide families iturin A and fengycin were found to correlate positively with extract inhibition, but a peak with unidentified chemical composition (designated as peak 48) showed the strongest correlation with extract inhibition. HPLC data were used to construct models for the production of iturin A, fengycin, and peak 48 as a function of the substrate moisture content, incubator temperature, and aeration rate in the solid-state bioreactors. Maximum production of all compounds occurred at the highest moisture content (1.7 g/g dry basis) and lowest incubator temperature (19 degrees C) tested. Optimal aeration rates for the production of the two known lipopeptides and peak 48 were 0.1 and 1.5 L/min, respectively.     

Insecticidal and antifungal activity of a protein from Pouteria torta seeds with lectin-like properties

This paper describes the purification and characterization of a novel protein from the seeds of Pouteria torta (family Sapotaceae). The protein was purified by a combination of gel filtration, ion-exchange, and reverse phase chromatographies. SDS-PAGE of the purified protein resulted in a single protein band of 14 kDa in the presence and absence of DTT. The lectin-like activity of pouterin was best inhibited by glycoproteins such as fetuin, asialofetuin, heparin, orosomucoid, and ovoalbumin. Pouterin inhibited the growth of the fungi Fusarium oxysporum and Colletotrichum musae and of the yeast Saccharomyces cerevisiae. The incorporation of pouterin into an artificial diet (final concentration = 0.12%, w/w) caused 50% mortality in larvae of the insect Callosobruchus maculatus, whereas 0.08% pouterin produced an ED50.     

Characterization of volatile constituents of Haplopappus greenei and studies on the antifungal activity against phytopathogens

Essential oil of Haplopappus greenei A. Gray was obtained by hydrodistillation of aerial parts, which were subsequently analyzed by gas chromatography and gas chromatography-mass spectrometry. Major components were identified as carvacrol (8.7%), beta-pinene (7.6%), trans-pinocarveol (6.2%), and caryophyllene oxide (5.8%), respectively. In total, 104 components representing 84.9% of the investigated essential oil were characterized. Furthermore, the essential oil was evaluated for antimalarial, antimicrobial, and antifungal activities. However, only antifungal activity was observed against the strawberry anthracnose-causing fungal plant pathogens Colletotrichum acutatum, Colletotrichum fragariae, and Colletotrichum gloeosporioides using the direct overlay bioautography assay. Major essential oil components were also evaluated for antifungal activity; the carvacrol standard demonstrated nonselective activity against the three Colletotrichum species and the other compounds were inactive.     

Assessment of the antibacterial activity and the antidiarrheal function of flavonoids from bayberry fruit

Chinese bayberry fruits are used as a folk medicine to cure diarrhea. However, the active compounds have not yet been reported. We found that bayberry fruit extract showed significant antibacterial activity against Salmonella, Listeria, and Shigella, and the minimal inhibitory concentration (MIC) ranged from 2.07 to 8.28 mg/mL. Positive relationships were found between the antibacterial activity and the total polyphenol (r = 0.88) and flavonoid contents (r = 0.92) of samples using different extraction times. The active compounds showed green or blue fluorescence under UV light using the bioautography method and were purified using a polyamide column. The fraction F1 with the most activity was comprised of flavonoids, which included cyanidin-3-O-glucoside, myricetin deoxyhexoside, quercetin-3-O-glucoside, and quercetin deoxyhexoside, and it also possessed an antidiarrheal activity (p < 0.10) at 80 mg/kg in mice. These findings provide scientific evidence for the antidiarrheal function of bayberry.     

Use of a substrate/alliinase combination to generate antifungal activity in situ

Allicin, an active ingredient of garlic, possesses a range of antimicrobial properties. Unfortunately, certain properties of the compound, such as chemical instability and low miscibility with water, have hampered its practical use in the past. Here, we show that it is possible to use a binary system consisting of the plant enzyme alliinase and its substrate alliin to generate allicin, and hence antifungal activity, in situ. During application, the two inactive components generate compounds that inhibit growth and infection-related development of the rice blast fungus Magnaporthe grisea. It is therefore possible to "trigger" biological activity in a controlled, yet effective manner. Apart from circumventing many of the drawbacks of allicin, this binary system has additional important advantages, such as low toxicity of its individual components and selective activation. Importantly, alliinase is also able to use different substrates, therefore paving the way to a range of novel, binary antimicrobial systems with custom-made chemical and biochemical properties.     

Phytoalexins from the Vitaceae: biosynthesis, phytoalexin gene expression in transgenic plants, antifungal activity, and metabolism

Resistance of plants to infection by phytopathogenic microorganisms is the result of multiple defense reactions comprising both constitutive and inducible barriers. In grapevine, the most frequently observed and best characterized defense mechanisms are the accumulation of phytoalexins and the synthesis of PR-proteins. Particular attention has been given here to stilbene phytoalexins produced by Vitaceae, specifically, their pathway of biosynthesis (including stilbene phytoalexin gene transfer experiments to other plants) and their biological activity together with fungal metabolism.     

Composition and antibacterial activity of Pseudocytisus integrifolius (Salisb.) essential oil from Algeria

The essential oil composition of an endemic Algerian Cruciferae, Pseudocytisus integrifolius (Salisb.) Rehder, was analyzed by gas chromatography (GC) and GC-mass spectrometry (MS). Eighty-three components representing more than 96.5% of the oil were identified. The major components were dimethyl disulfide (33.4%), dimethyl trisulfide (24.2%), and an unsaturated nitrile (31.7%). Fractionation on a silica gel column led to the identification of trace-level compounds, in particular, polar compounds such as nitriles and aldehydes, and to the isolation of dimethyl disulfide, dimethyl trisulfide, and an unsaturated nitrile. Structural analysis using high-resolution mass spectrometry (HRMS) and 1H,13C NMR techniques enabled the identification of pent-4-enenitrile. Variation in essential oil composition and yields was studied according to harvesting time, drying, and parts of the plant. The essential oil of aerial parts was tested for its antibacterial activity using a paper disk method. The oil was effective on the inactivation of Escherichia coli and Pseudomonas aeruginosa and ineffective on the inactivation of Staphylococcus aureus.     

Composition and antifungal activity on soil-borne pathogens of the essential oil of Salvia sclarea from Greece

The hydrodistilled essential oils of the aerial parts of wild-growing Salvia sclarea originated from two localities in Greece were analyzed by GC-MS. Sixty-six compounds, representing 93.26-98.19% of the oils, were identified. Linalyl acetate (19.75-31.05%), linalool (18.46-30.43%), geranyl acetate (4.45-12.1%), and alpha-terpineol (5.08-7.56%) were the main components. The antifungal activity of the oil of one locality and of the main components, linalyl acetate and linalool, was evaluated in vitro against three soil-borne pathogens.     

Isolation, characterization and antifungal activity of major constituents of the Himalayan lichen Parmelia reticulata Tayl

Antifungal activity of hexane, ethyl acetate and methanol extracts of Parmelia reticulata was evaluated against soilborne pathogenic fungi, namely, Sclerotium rolfsii, Rhizoctonia solani, R. bataticola, Fusarium udum, Pythium aphanidermatum and P. debaryanum by poisoned food technique. Maximum antifungal activity was exhibited by hexane and ethyl acetate extracts against most of the test pathogens. Secondary metabolites, namely, (±)-isousnic acid, (±)-protolichesterinic acid, atranorin, evernyl, ethyl hematommate, ethyl orsellinate, methyl hematommate (3-formyl-2,4-dihydroxy-6-methylbenzoic acid methyl ester), 2-hydroxy-4-methoxy-3,6-dimethylbenzoic acid, 1-hydroxy-3,6-dimethoxy-8-methyl-xanthen-9-one, baeomycesic acid and salazinic acid, were isolated from the above extracts and identified by 1H NMR, 13C NMR and mass spectroscopic methods. When these metabolites were tested for antifungal activity against test pathogens, maximum antifungal activity was exhibited by (±)-protolichesterinic acid against R. solani (ED50=23.09 μg mL(-1)) and P. debaryanum (ED50=16.07 μg mL(-1)) and by atranorin against S. rolfsii (ED50=39.70 μg mL(-1)). The antifungal activity of protolichesterinic acid was found to be comparable to that of hexaconazole, a commercial fungicide.     

Isolation and antifungal activity of 4-phenyl-3-butenoic acid from Streptomyces koyangensis strain VK-A60

An antifungal compound was isolated from the culture broth of Streptomyces koyangensis strain VK-A60 using various chromatographic procedures. On the basis of the high-resolution EI-mass and 1H and 13C NMR data, the compound was identified as 4-phenyl-3-butenoic acid. Colletotrichum orbiculare, Magnaporthe grisea, and Pythium ultimum were most sensitive to 4-phenyl-3-butenoic acid. Strong inhibitory effects of 4-phenyl-3-butenoic acid also were found against Pectobacterium carotovorum subsp. carotovorum and Ralstonia solanacearum. 4-Phenyl-3-butenoic acid effectively suppressed the development of M. grisea on rice leaves at the concentration of more than 10 microg/mL, and the protective activity was in general similar to that of the commercial fungicide tricyclazole. Treatment with 100 microg/mL of 4-phenyl-3-butenoic acid also effectively inhibited the anthracnose development on cucumber plants, although its in vivo efficacy was somewhat less effective than that of the commercial fungicide chlorothalonil.     

Structure elucidation and antifungal activity of an anthracycline antibiotic, daunomycin, isolated from Actinomadura roseola

The actinomycete strain Ao108 producing antifungal metabolites active against some plant pathogenic fungi was identified as Actinomadura roseola, based on the analyses of morphological and physiological characteristics. The antibiotic Da2B that showed a strong antifungal activity was isolated from the culture broth and mycelial mats of A. roseola strain Ao108 using various chromatographic procedures. On the basis of (1)H NMR, (13)C NMR, and 2-D NMR correlation data, the antibiotic Da2B was confirmed to have the structure of an anthracycline antibiotic, daunomycin. In vitro antimicrobial spectrum tests showed that the antibiotic Da2B had substantial inhibitory activity (10 microg mL(-)(1) of MICs) against mycelial growth of Phytophthora capsici and Rhizoctonia solani. The antibiotic also showed antiyeast activity against Saccharomyces cerevisiae, but the growth of Candida albicans was not affected. Antibacterial activity was found only against Gram-positive bacteria. In the further evaluation of in vivo efficacy, application of the antibiotic Da2B effectively inhibited the development of Phytophthora blight in pepper plants. However, the control efficacy of the antibiotic against Phytophthora infection was somewhat less than that of metalaxyl. The antibiotic Da2B did not show any phytotoxicity on pepper plants even at 500 microg mL(-)(1).     

Chemical composition and antifungal activity of Arnica longifolia, Aster hesperius, and Chrysothamnus nauseosus essential oils

Essential oils from three different Asteraceae obtained by hydrodistillation of aerial parts were analyzed by gas chromatography (GC) and gas chromatography-mass spectrometry (GC/MS). Main compounds obtained from each taxon were found as follows: Arnica longifolia carvacrol 37.3%, alpha-bisabolol 8.2%; Aster hesperius hexadecanoic acid 29.6%, carvacrol 15.2%; and Chrysothamnus nauseosus var. nauseosus beta-phellandrene 22.8% and beta-pinene 19.8%. Essential oils were also evaluated for their antimalarial and antimicrobial activity against human pathogens, and antifungal activities against plant pathogens. No antimalarial and antimicrobial activities against human pathogens were observed. Direct bioautography demonstrated antifungal activity of the essential oils obtained from three Asteraceae taxa and two pure compounds, carvacrol and beta-bisabolol, to the plant pathogens Colletotrichum acutatum, C. fragariae and C. gloeosporioides. Subsequent evaluation of antifungal compounds using a 96-well micro-dilution broth assay indicated that alpha-bisabolol showed weak growth inhibition of the plant pathogen Botrytis cinerea after 72 h.