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A novel 2,3-epoxy naphthoquinol, named (6R,7R,8R)-theissenone A (1), possessing an oxatricyclo[5.4.0.03,5]undeca-trien-2-one skeleton, together with two known compounds, (6S,7R,8R)-theissenone (2) and arthrinone (3), were produced by an endophytic fungus, Arthrinium marii M-211, which was isolated from mangrove plants. The structure of 1, including the absolute stereochemistry, was elucidated by analysis of nuclear magnetic resonance (NMR) and mass spectrometry (MS) data and time-dependent density functional theory (TDDFT) calculations of electronic circular dichroism (ECD) spectra. Additionally, the absolute structure of 2 was deduced as a diastereomer of 1 using ECD spectral data analysis. Compounds 1, 2 and 3 exhibited cytotoxic activity against the H4IIE rat hepatoma cells, with IC50 values of 67.5, 46.6 and 13.4 µM, respectively.

Amomum compactum Soland Ex Maton is one of the Zingiberaceae family plants which is the endemic plants from West Java, Indonesia. This study was aimed to determine the chemical structure of flavonoid compounds from n-hexane extract of A.compactum Sol. Ex Maton roots. Dried powder of the roots was extracted consecutively with n -hexane, ethyl acetate, and methanol solvents. Three flavonoids, 5-hydroxy-3,7,4`-trimethoxy kaempferol ( 1 ), 5-hydroxy-3,7,3',4'-tetra methoxy kaempferol ( 2) and 4'-hydroxy-3,5,7-trimethoxy kaempferol ( 3 ), have been isolated from the roots of A. compactum Sol. Ex Maton. The chemical structures of compounds 1 - 3 were identified by spectroscopy data including infrared 1D-NMR, 2D-NMR and HRTOF-MS as well as by comparison with previously reported spectral data. Compounds 1-3 were isolated from this plant for the first time and showed free radical DPPH scavenging activity.

In this study, we report the identification and characterization of an acetyl xylan esterase, designated as AoAXEC, which was previously annotated as a hypothetical protein encoded by AO090023000158 in the Aspergillus oryzae genomic database. Based on its amino acid sequence, a low sequence identity to known acetyl xylan esterases was observed in the sequence of characterized acetyl xylan esterase. The gene fused with α-factor signal sequence of Saccharomyces cerevisiae instead of the native signal sequence was cloned into a vector, pPICZαC, and expressed successfully in Pichia pastoris as an active extracellular protein. The purified recombinant protein had pH and temperature optima of 7.0 and 50 °C, respectively, and was stable up to 50 °C. The optimal substrate for hydrolysis by the purified recombinant AoAXEC, among a panel of α-naphthyl esters (C2–C16), was α-naphthyl propionate (C3), with an activity of 0.35 ± 0.006 units/mg protein. No significant difference of the Km value was observed between C3 (2.3 ± 0.7 mM) and C2 (1.9 ± 0.4 mM). In contrast, kcat value for C3 (18 ± 3.9 s−1) was higher compared to C2 (4.5 ± 0.7 s−1). The purified recombinant enzyme displayed a low activity toward acyl chain substrates containing eight or more carbon atoms. Recombinant AoAXEC catalyzed the release of acetic acid from wheat arabinoxylan. However, no activity was detected on methyl esters of ferulic, p-coumaric, caffeic, or sinapic acids. Additionally, the liberation of phenolic acids, such as ferulic acid, from wheat arabinoxylan was not exhibited by the recombinant protein.

Three new β-resorcylic acid derivatives, 1–3, were obtained from the endophytic strain Lasiodiplodia theobromae GI-1005, which was isolated from the mangrove plant Rhizopora mucronata. The structures of new compounds together with five known compounds were elucidated by NMR and MS analyses, and chemical means. The isolated compounds were evaluated for phytotoxicity against Digitaria ciliaris (southern crabgrass) and Lactuca sativa (lettuce). Compound 1 showed growth inhibitory effects against D. ciliaris in a dose-dependent manner.

A new alpha-pyrone, asperpyrone (1) and two known compounds, stigmasterol (2) and 7-hydroxy-3-(2,3-dihydroxybutyl)-1(3H)-Isobenzofuranone (3) were isolated from the solid rice culture of the endophytic fungus Aspergillus sp., an endophytic fungus isolated from the fresh inner tissue of the barks of Garicinia smeathmannii. Their structures were determined by extensive spectral analysis including 1D and 2D NMR, HRESIMS and single-crystal X-ray crystallographic analysis. Asperpyrone (1) was tested against an axenic culture of Entamoeba moshkovskii, but did not exhibit any significant amoebicidal activity.

Two new oleanane-type triterpene saponins, named polythosides A and B (1 and 2), together with a known compound, silphioside E (3), were isolated from the roots of Acacia polyacantha. Their structures were elucidated by analysis of 1D and 2D-NMR experiments, and mass spectrometry (HR-ESITOF-MS) as oleanolic acid 3-O-β- d -galactopyranosyl-(1→2)-β- d -xylopyranosyl-(1→6)-[β- d -xylopyranosyl-(1→4)]-[β- d -xylopyranosyl-(1→2)]-β- d -galactopyranoside (1), 3-O-[β- d -galactopyranosyl-(1→2)-β- d -xylopyranosyl-(1→6)-[β- d -xylopyranosyl-(1→4)]-[β- d -xylopyranosyl-(1→2)]-β- d -galactopyranosyl] oleanolic acid-28-O-β- d -glucopyranosyl ester (2) and 3-O-β- d -glucopyranosyl-(1→2)-β- d -glucopyranosyl] oleanolic acid 28-O-β- d -glucopyranosyl ester (3). The cytotoxic effect of isolated saponins was evaluated on the H4IIE rat hepatoma cell line. The results show that none of the compounds (assayed at 100 μM) showed cytotoxicity against H4IIE cells.

Three new metabolites, including two alkaloids, vancouverone A (1) and vancouverone B (2), one α-pyrone, 10,11-dihydroxy-citreoviridin (3), and two known compounds, neocitreoviridinol (4) and PF1140 (5), were isolated from Penicillium vancouverense YY-1 derived from gastric flora of the sea snail Monodonta neritoides. Their structures and configurations were elucidated using spectroscopic analyses. Compound 2 exhibited phytotoxicity against plant species (Lactuca sativa, Trifolium repens, Raphanus sativus var. longipinnatus, and Brassica rapa var. perviridis).

A seco-apotirucallane-type triterpenoid, namely angustifolianin (1), along with three dammarane-type triterpenoids, (20S, 24S)-epoxy-dammarane-3β,25-diol (2), 3-epi-cabraleahydroxylactone (3), and ...

Six new sesquiterpenes, tsukiyols A-C, neoilludin C, and 4-O-methylneoilludins A and B, were isolated from the fruiting body of Omphalotus japonicus (Kawam.) Kirchm. & O. K. Mill. Additionally, six known compounds, illudin S, neoilludins A-B, 5-hydroxydichomitol, ergosterolperoxide, and 3β,5α,9α-trihydroxyergosta-7,22-diene-6-one, were also obtained. Their chemical structures were determined with MS, IR, and NMR spectra and the absolute configurations of neoilludins A-C, 4-O-methylneoilludins A, and B were determined with electronic circular dichroism (ECD). Illudin S and 3β,5α,9α-trihydroxyergosta-7,22-diene-6-one showed cytotoxicity against human acute promyelocytic leukemia HL60 cells. Illudin S, 4-O-methylneoilludin A, B, and tsukiyol C showed growth-restoring activity against mutant yeast via Ca2+-signal transduction.

Three new limonoids namely pentandricines B–D (1–3), were isolated from n-hexane extract of the stem bark of Chisocheton pentandrus, along with four known limonoids (4-7). The chemical structures of pentandricines B–D, were fully elucidated based on UV, IR, HR-TOFMS, 1D- and 2D-NMR spectra. Furthermore, compounds 1-7 were evaluated for their cytotoxic activity against MCF-7 breast cancer cells in vitro, with compound 7 (nimonol) showing strongest cytotoxicity activity with an IC50 value of 22.03 ± 0.026 μM.

A tannase-encoding gene, AotanB, from Aspergillus oryzae RIB40 was overexpressed in A. oryzae AOK11 niaD-deficient mutant derived from an industrial strain under the control of an improved glucoamylase gene promoter PglaA142. The recombinant tannase, designated as rAoTanBO, was produced efficiently as an active extracellular enzyme. Purified rAoTanBO showed a smeared band with a molecular mass of approximately 80–100 kDa on sodium dodecyl sulfate polyacrylamide gel electrophoresis. The rAoTanBO had a molecular mass of 65 kDa, after treatment with endo-β-N-acetylglucosaminidase H. Purified rAoTanBO exhibited maximum activity at 30–35°C and pH 6.0. The tannase activity of purified rAoTanBO towards natural and artificial substrates was 2–8 folds higher than that of the recombinant enzyme produced by Pichia pastoris, designated as rAoTanBP. N-terminus of the mature rAoTanBP had six more amino acids than the N-terminus of the mature rAoTanBO. Kinetic analyses showed that rAoTanBO had higher catalytic efficiency (kcat/Km) than rAoTanBP. rAoTanBO was stable up to 60°C and higher thermostability than rAoTanBP. N-linked oligosaccharides had no effect on the activity and stability of rAoTanBO and rAoTanBP.

Phytochemical investigation led to the isolation of three alkaloids [skimmianine (1), vulcanine (2) and evoxine (3)] from the aerial parts of Haplophyllum tuberculatum, and two diacetylenic spiroketal enol ethers [spiroketalenolether polyyne (4) and [7-hexa-2,4-diynylidene-1,6-dioxaspiro[4.4]nona-2,8-dien-4-yl] acetate (5)] and a sesquiterpene lactone [8α acetoxy-11β,13dihydrokauniolide (6)] from the aerial parts of Chrysanthemum coronarium. The chemical structures of the isolated compounds were determined by using spectral data of 1H NMR, 13C NMR, as well as by comparison with literature data. Compounds 4, 1 and 5 were the most potent inhibitors against plant pathogenic bacteria Rhizobium radiobacter, Ralstonia solanacerum and Pectobacterium carotovorum ss. carotovorum with minimum inhibitory concentration (MIC) values of 31.3, 62.5 and 31.3 µg/ml, respectively. In antifungal assay, compounds 1 (MIC 62.5 µg/ml), 2 (MIC 62.5 µg/ml) and 5 (MIC 125 µg/ml) caused the highest inhibitory effects against phytopathogenic fungi Verticillium dahliae, Fusarium oxysporum and Alternaria solani, respectively. Moreover, crude extract of H. tuberculatum and 4 were significantly the most potent inhibitors of seed germination, root and shoot growth of Echinochloa crus-galli.

In the course of our continuing search for anticancer compounds from Chisocheton species, three steroids, stigmast-5-en-3β-ol ( 1 ), stigmast-5-en-3β-ol-3- O -β- D -glucopyranoside ( 2 ) and stigmast-5,22-dien-3β-ol-3- O -β- D -glucopyranoside ( 3 ), were obtained from the stembark of Chisocheton celebicus . The structures of compound 1 - 3 were identified with spectroscopic data including IR, 1D-NMR, 2D-NMR and TOF-MS, as well as by comparing with those spectral data previously. Compounds 1 - 3 , were evaluated for their cytotoxic effects against P-388 murine leukemia cells and displayed the cytotoxicity activity with IC 50 values of 12.45 ± 0.050, 52.27 ± 0.031 and 62.52 ± 0.076 µg/mL, respectively.

The endophytic fungus, Clonostachys rosea B5-2 was isolated from mangrove plants and subjected to the one strain many compounds (OSMAC) methodology. By this approach, it was found that modification...

Two new naphthoquinone derivatives, 6-hydroxy-astropaquinone B (1) and astropaquinone D (2) as well as the known compound 3-O-methyl-9-O-methylfusarubin (3) were isolated from Fusarium napiforme, an endophytic fungus isolated from the mangrove plant, Rhizophora mucronata. The structures of 1 and 2 were determined by 1D and 2D NMR spectroscopic analyses. Compounds 1, 2 and 3 exhibited moderate antibacterial activity against Staphylococcus aureus and Pseudomonas aeruginosa. Furthermore, 1, 2 and 3 were phytotoxic action in lettuce seeding at a concentration of 30 μg · mL−1.

Fractionation of ethyl acetate extract obtained by culturing the endophytic fungus Pseudocosmospora sp. Bm-1-1 resulted in the isolation of four new meroterpenoids, cosmosporin A (1), 6-carboxy-cosmosporin A (2), rel-(6aS,10aR)-Δ9-tetrahydrocannabiorcolic acid B (3) and 8′-hydroxy-cannabiorcichromenic acid (4), in addition to four known compounds 5–8. Structures were elucidated by spectral analysis, as well as by directly comparing the spectral data of new compounds with those of known compounds. Cannabiorcichromenic acid (5), decarboxy-cannabiorcichromenic acid (6), and rel-(6aS,10aR)-decarboxy-Δ9-tetrahydrocannabiorcolic acid B (8) restored growth of a Saccharomyces cerevisiae mutant strain involving Ca2+ signal transduction. Furthermore, compounds 3 and 8 had cytotoxic activity against HL60 cells (3: IC50 = 24.1 μM and 8: IC50 1.6 μM).

Four new triterpenoids, namely, chisopaten A-D (1-4), were isolated from the n-hexane extract of the bark of Chisocheton patens Blume. The chemical structures of new compounds were elucidated on the basis of spectroscopic data interpretation. All isolated compounds were evaluated for their cytotoxic activity against MCF-7 breast cancer lines. Chisopaten A and C, showed strongest cytotoxicity activity with IC50 values of 4.01 ± 0.008 and 4.33 ± 0.009 μM, respectively.

Five polyketides, paralactonic acids A-E (1-5) were isolated from Paraconiothyrium sp. SW-B-1, an endophytic fungus isolated from the seaweed, Chondrus ocellatus Holmes. Their structures were determined by various spectroscopic methods, predominantly by 2D NMR spectroscopic analyses. The absolute configurations of compounds 1-5 were determined by comparison of the experimental and calculated ECD spectra. Compound 5 showed moderate antibacterial activity and restored the growth of a mutant yeast strain inhibited by hyperactivated Ca2+-signaling.

Nine new β-resorcylic acid derivatives, (15S)-de-O-methyllasiodiplodin (1), (13S,15S)-13-hydroxy-de-O-methyllasiodiplodin (2), (14S,15S)-14-hydroxy-de-O-methyllasiodiplodin (3), (13R,14S,15S)-13,14-dihydroxy-de-O-methyllasiodiplodin (4), ethyl (S)-2,4-dihydroxy-6-(8-hydroxynonyl)benzoate (5), ethyl 2,4-dihydroxy-6-(8-hydroxyheptyl)benzoate (6), ethyl 2,4-dihydroxy-6-(4-methoxycarbonylbutyl)benzoate (7), 3-(2-ethoxycarbonyl-3,5-dihydroxyphenyl)propionic acid (8), and isobutyl (S)-2,4-dihydroxy-6-(8-hydroxynonyl)benzoate (9), together with a known ethyl 2,4-dihydroxy-6-(8-oxononyl)benzoate (10) were obtained from Lasiodiplodia theobromae GC-22. The structures of these compounds were elucidated by extensive spectroscopic analyses. Compounds 1, 3, and 6 showed growth inhibitory effects against Digitaria ciliaris. Conversely, treatment with compounds 5, 6, 7, 9, and 10 stimulated elongation activity toward the root of Lactuca sativa. These data expand the repertoire of new β-resorcylic acid derivatives that may function as lead compounds in the synthesis of new agrochemical agents.

From the crude extract of the plant-associated fungus Xylaria sp. collected in Cameroon, a new 2 H-chromene derivative, hexacycloxylariolone (1), and a new polyalcohol, xylatriol (2), were isolated, in addition to 3 known compounds, 2,3-furandiol (3), 1,8-dimethoxynaphthalene (4), and 1-palmitoyl-rac-glycerol (5). Their chemical structures were established on the basis of the interpretation of spectroscopic data. Hexacycloxylariolone (1), 1,8-dimethoxynaphthalene (4), and 1-palmitoyl-rac-glycerol (5) showed antiproliferative activity by inhibiting the growth of Raw 264.7 and THP-1 cancer cell lines.

Eleven undescribed triterpenoids (pentandrucines A to K) were isolated from the n-hexane extract of the stem bark of Chisocheton pentandrus (Blanco) Merr. These comprised ten undescribed dammarane-type triterpenoids and one undescribed apotirucallane-type triterpenoid. Additionally, two dammarane-type triterpenoids, four apotirucallane-type triterpenoids and two tirucallane-type triterpenoids were also isolated. The chemical structures of pentandrucine A-K, were fully elucidated using 1D and 2D-NMR, and high resolution MS. All of the compounds were evaluated for cytotoxic activity against MCF-7 breast cancer cells in vitro. Melianodiol proved to be the most active with an IC50 of 16.84 μM comparing favourably with Cisplatin (13.2 μM).

The rutinosidase (Rut)-encoding gene Aorut has been expressed in Pichia pastoris with its native signal sequence from Aspergillus oryzae. Biochemical and structural investigation of the purified recombinant mature A. oryzae Rut (AoRut), designated rAoRutM, was performed in this study. A 1.7-A resolution crystal structure of rAoRutM was determined, which is an essential step forward in the utilization of AoRut as a potential catalyst. The crystal structure of rAoRutM was represented by a (β/α)8 TIM barrel fold with structural similarity to that of rutinosidase from Aspergillus niger (AnRut) and an exo-β-(1,3)-glucanase from Candida albicans. The crystal structure revealed that the catalytic site was located in a deep cleft, similarly to AnRut, and that internal cavities and water molecules were also present. Purified rAoRutM hydrolyzed not only 7-O-linked and 3-O-linked flavonoid rutinosides but also 7-O-linked and 3-O-linked flavonoid glucosides. rAoRutM displayed high catalytic activity toward quercetin 3-O-linked substrates such as rutin and isoquercitrin, rather than to the 7-O-linked substrate, quercetin-7-O-glucoside. Unexpectedly, purified rAoRutM exhibited increased thermostability after treatment with endo-β-N-acetylglucosaminidase H. Circular dichroism (CD) spectra of purified intact rAoRutM and of the enzyme after N-deglycosylation showed a typical α-helical CD profile; however, the molar ellipticity values of the peaks at 208 nm and 212 nm differed. The Km and kcat values for the substrates modified by rutinose were higher than those for the substrates modified by β-d-glucose. IMPORTANCE Flavonoid glycosides constitute a class of secondary metabolites widely distributed in nature. These compounds are involved in bitter taste or clouding in plant-based foods or beverages, respectively. Flavonoid glycoside degradation can proceed through two alternative enzymatic pathways: one that is mediated by monoglycosidases and another that is catalyzed by a diglycosidase. The present report on the biochemical and structural investigation of A. oryzae rutinosidase provides a potential biocatalyst for industrial applications of flavonoids.

A novel lipolytic enzyme-encoding gene, lipO745, from Aspergillus oryzae RIB40 was cloned and expressed in Pichia pastoris. Purified recombinant LipO745 (rLipO745) had a molecular mass of approximately 60 kDa on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. rLipO745 exhibited maximum activity at 40 °C and pH 7.0 and was stable at temperatures ≤ 40 °C. The substrate specificity of purified rLipO745 was analyzed using α-naphthyl esters as artificial substrates and various triacylglycerol and sterol esters as natural substrates. From among a panel of α-naphthyl esters (C2-C16), α-naphthyl butyrate (C4), with an activity of 269 ± 3.3 units/mg protein, was the optimal substrate for hydrolysis by the purified recombinant protein. The Km and kcat values of rLiO745 for the C4 substrate were 0.073 ± 0.0012 mM and 608 ± 108 s-1, respectively. The purified recombinant enzyme had considerable hydrolytic activity toward tributyrin, tripalmitin, and triolein, indicating lipase activity, and toward cholesteryl acetate, butyrate, palmitate, and oleate, indicating sterol esterase activity. Transesterification activities between tributyrin and cholesterol or between tributyrin and campesterol were also determined.

A new propylcholesterol-type steroid, namely (22E,24S)-24-propylcholest-5en-3α-acetate (1), has been isolated from the stembark of Aglaia angustifolia (Miq.). The structure of 1 was determined on the basis of spectroscopic data including 1D- and 2D-NMR as well as high resolution mass spectroscopy analysis. Compound 1 showed weak activity against the MCF-7 breast cancer cell line.

Two new compounds, a new lactone, quercilactone A (1), and (17R)-hydroxynafuredin (5), as well as five known compounds, scytalone (2), 3S,4R-hydoxy-scytalone (3), nafuredin (4), (+)-(3R,5R)-3-hydroxy-5-decanolide (6) and 3-ethyl-4-hydroxy-6-methyl-2H-pyran-2-one (7), were isolated from Raffaelea quercivora, a fungus that causes Japanese oak wilt disease. The structures of these compounds were determined by 1D and 2D NMR spectroscopic analyses. The absolute configuration at C-17 of 5 was determined to be R by the modified Mosher’s method. Compounds 1, 2, and 7 exhibited weak phytotoxic activity in lettuce seedlings at a concentration of 100 μg mL−1.

Twenty-eight compounds were isolated from endophytic, soil and marine fungi and their structures were elucidated through spectroscopic methods. The isolated compounds were tested for their antibacterial and herbicidal activities against phytopathogenic bacteria and barnyard grass weed for the first time. Methyleurotinone (14) was the most potent compound against Pectobacterium carotovorum subsp. carotovorum, Pseudomonas syringae pv. syringae, Rhizobium radiobacter and Ralstonia solanacearum with minimum inhibitory concentration (MIC) values of 31.3, 125, 31.3 and 125 mg/L, respectively. Compounds 13–15 were highly effective in reducing the development of potato tuber soft rot disease caused by P. carotovorum subsp. carotovorum. Furthermore, twelve of the tested compounds induced a significant reduction in seed germination of Echinochloa crus-galli at 2 mM with compounds 8 and 26 causing complete inhibition of seed germination. Also, compounds 4, 22, 5, 8, 18 and 25–27 induced remarkable reduction of root and shoot growth of E. crus-galli at 2 mM.

We report the characterization of tannase-encoding gene, AotanB, from Aspergillus oryzae and its recombinant enzyme expressed in Pichia pastoris. The gene except for the signal sequence was cloned into a vector pPICZαA and the recombinant protein was secreted into the medium as an active enzyme. Recombinant AoTanB highly expressed in the incubation at 18°C compared to 30°C. Purified recombinant protein exhibited smeared band with molecular mass of approximately 90–120 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The recombinant protein yielded molecular mass of 65 kDa after N-deglycosylation. Purified recombinant enzyme had a pH and a temperature optima of 6.0 and 30–35°C, respectively, and was stable up to 40°C. Recombinant AoTanB was able to release gallic acid from natural substrates, such as (−)-catechin gallate, (−)-epicatechin gallate, (−)-gallochatechin gallate, and (−)-epigallocatechin gallate. The enzyme also hydrolyzed ethyl protocatechuate. Meanwhile, no activity was detected toward ethyl 4-hydroxybenzoate. The activity of recombinant AoTanB was lower toward natural substrates compared to that of AoTanA from A. oryzae. The lower catalytic efficiency (kcat/Km value) toward ethyl protocatechuate was due to a combination of increased Km and considerably decreased kcat. Kinetic analysis of the recombinant AoTanB showed that kcat values toward natural substrates decreased compared to those of recombinant AoTanA. Therefore, recombinant AoTanB showed a decrease in catalytic efficiency (kcat/Km value) compared to recombinant AoTanA was due to considerably lower kcat value.

Flavonoid glycoside degradation can proceed through two alternative enzymatic pathways: one that is mediated by monoglycosidases, and the other catalyzed by a diglycosidase. β-Diglycosidase performs the flavonoid deglycosylation in a single reaction. The characterized β-diglycosidase activities recognize the following disaccharidic sugar moieties: β-primeverose, acuminose, vicianose, and β-rutinose. The present paper reviews the biochemical characteristics and potential industrial applications of microbial β-diglycosidases that break down plant diglycoconjugated flavonoids.

A new pyrrocidine A derivative, 19-epi-pyrrocidine A (1) was isolated from the insect–derived fungus Neonectria ramulariae In-2, together with known pyrrocidine A (2). The structure of 1 was determined by extensive 1D- and 2D-NMR, and MS spectral analyses. The absolute configuration of 2 was confirmed for the first time by X-ray crystallography. Compound 1 exhibited cytotoxicity against human acute promyelocytic leukemia HL60 cells (IC50 2.05 μM). In addition, semi-synthetic compounds 19-O-methyl-pyrrocidine A (3), 19-O-acetyl-pyrrocidine A (4), and 1,2-4,5-17,18-triepoxy-pyrrocidine A (5) were found to possess significant cytotoxicity, with IC50 values of 0.66 μM, 0.19 μM, and 2.00 μM, respectively.

Six flavonoid compounds, kaempferol (1), quercetin (2), acacetin (3), quercetin-3- O - β -D-glucopyranoside (4), kaempferol-3- O - β -D-glucopyranoside (5), and isorhamnetin-3- O - β -D-glucopyranoside (6), were isolated from the methanol extract of the Lygodium microphyllum herb. The chemical structures of compounds 1-6 were identified on the basis of spectroscopic evidence and comparison with previously reported spectral data. Compounds 1-6 were isolated for first time from this plant. Their antioxidant activity against DPPH (2,2-Diphenyl-1-picrylhydrazyl) was evaluated. Among the compounds, quercetin (2) showed high antioxidant activity with IC 50 values of 6.94 ± 0.03 μ g/mL.

Two dammarane-type triterpenoids, 20 S ,24 S -epoxy-3α,25-dihydroxydammarane ( 1 ) and 3α-acetyl-20 S ,24 S -epoxy-3α,25-dihydroxydammarane ( 2 ), have been isolated from the stembark of Aglaia argentea . The chemical structure of compounds ( 1 and 2 ) were identified by spectroscopic evidences including UV, IR, 1D-NMR, 2D-NMR and MS as well as by comparing with previously reported spectral data. Those compounds were isolated from this plant for first time. Compounds ( 1 and 2 ) showed cytotoxic activity against P-388 murine leukemia cells with IC 50 values of 23.96 and 8.14 mM, respectively. DOI: http://dx.doi.org/10.15408/jkv.v4i1.7065

Three previously undescribed oleanane-type saponins acylated with monoterpenic acid, kamerunosides A, B, and C were isolated from the stem liana of Acacia kamerunensis . Their structures were elucidated on the basis of extensive 1D and 2D NMR data and HRESIMS analyses. Their cytotoxicities were evaluated against HL60 cells. Kamerunoside B was the most cytotoxic.

Basella alba is a perennial plant of the Basellaceae and is known by various common names including Malabar spinach. There are few insects that cause damage to B. alba. In this study, we examined the effect of B. alba leaves on the growth of Spodoptera litura larvae. B. alba leaves and a methanolic extract of the leaves inhibited the growth of S. litura larvae. Half of the larvae reared on the leaves died within 1 week. We found that two flavonoids, vitexin, and vitexin-2″-O-arabinofuranoside, were abundant in the methanol extract of leaves. When larvae were reared on purified vitexin or vitexin-2″-O-arabinofuranoside, their growth was significantly impaired compared with larvae reared on control spinach leaves. These results suggested that the flavonoid glycosides in B. alba leaves act as deterrents to S. litura larvae.

A new pregnane-type steroid, 2α-hydroxy-3α-methoxy-5α-pregnane (1), together with three known dammarane-type triterpenoid, 3β-acetyl-20S,24S-epoxy-25-hydroxydammarane (2), 20S,24S-epoxy-3α,25-dihydroxydammarane (3), and eichlerianic acid (4) have been isolated from the stem bark of Aglaia elliptica. The structure s were determined by spectroscopic methods including the 2D-NMR techniques. Compound 1-4 showed moderate cytotoxic activity against P-388 murine leukemia cells .

Three new meroterpenoid derivatives, furanocochlioqiunol (1) and furanocochlioqiunone (2), as well as nectrianolin D (3), together with two known biogenetically related compounds 4 and 5 were isolated from a mixed culture of two mangrove-derived fungi, Clonostachys rosea B5–2 and Nectria pseudotrichia B69–1. The structures of 1–3 were deduced based on the interpretation of HRMS and NMR data. Compounds 1–5 exhibited cytotoxicity against human promyelocytic leukemia (HL60) cells with IC50 values ranging from 0.47 to 10.16 μM.

800x600 Two mixture s of flavanoid compounds ( 1 and 2 ), a mixture of catechin ( 1a ) and epicatechin ( 1b ) , and a mixture of gallocatechin ( 2a ) and epigallocatechin ( 2b ) , were isolated from the active fraction of the stem bark of Aglaia elliptica methanol extract. The chemical structure of the compounds w as identified with spectroscopic data , including UV, IR, NMR ( 1 H, 13 C, DEPT 135°, HMQC, HMBC, 1 H- 1 H COSY) , and MS, and additionally compared with previously reported spectral data. All compounds were evaluated for their cytotoxic effects against P-388 murine leukemia cells. Compound 2 showed cytotoxicity against the P-388 murine leukemia cell , with an IC 50 value of 7.79 µg/mL, but compound 1 wa s found not to be active ( more than 100 µg/mL ) . Normal 0 false false false EN-ID X-NONE X-NONE

An α- l -rhamnosidase-encoding gene from Aspergillus oryzae, which belongs to the glycoside hydrolase family 78, was cloned and expressed in Pichia pastoris. SDS-PAGE of the purified recombinant α- l -rhamnosidase protein revealed smeared bands with apparent molecular mass of 90–130 kDa. After N-deglycosylation, the recombinant enzyme showed a molecular mass of 70 kDa. The enzyme exhibited optimal activity at a pH of 5.0 and a temperature of 70 °C. Specific activity of the enzyme was higher toward hesperidin than toward naringin, which consist of α-1,6 and α-1,2 linkages, respectively. The activity was also higher toward hesperidin than toward rutin, which consist of 7-O- and 3-O-glycosyl linkages of flavonoids, respectively. Kinetic analysis of the enzyme showed that the Michaelis constant (Km) was lowest toward rutin, moderate toward naringin, and higher toward p-nitrophenyl-α- l -rhamnopyranoside and hesperidin. Its high catalytic efficiency (kcat/Km) toward rutin was results of its low Km value while its high catalytic efficiency toward hesperidin was results of a considerably high kcat value.

The novel compound, 11-O-methylpetasitol (1), was isolated from Penicillium sp. N-175-1, and two new compounds, cosmochlorins D (5) and E (6), were isolated from Phomopsis sp. N-125. In addition, three known eremophilane sesquiterpenes, sporogen-AO1 (2), petasol (3) and 6-dehydropetasol (4), were isolated from Penicillium sp. N-175-1. The structures of 1, 5 and 6 were elucidated by a combination of extensive spectroscopic analyses, including 2D NMR, high-resolution electrospray ionization time-of-flight mass spectrometry (HRESITOFMS) and chemical reactions. Compounds 2, 3, 5 and 6 exhibited cytotoxicity to HL60 and 2 and 3 to HeLa cells. Furthermore, 2 and 3 showed robust growth-restoring activity of a Saccharomyces cerevisiae (cdc2-1 rad9Δ) mutant strain, whereas 5 and 6 exhibited minor growth-restoring activity in this strain. Thus, these compounds may inhibit the growth of HL60 and HeLa cells by blocking the cell cycle, and they may be utilized as new lead compounds that act as inhibitors of survival signal transduction pathways.

Two sesquiterpene-epoxycyclohexenone conjugates, nectrianolins A ( 1 ) and B ( 2 ), together with a sesquiterpene, nectrianolin C ( 3 ), were isolated from the brown rice culture of Nectria pseudotrichia 120-1NP, an endophytic fungus isolated from Gliricidia sepium . Their structures were determined on the basis of 1D-/2D-NMR spectroscopy and HRESIMS data analyses in combination with chemical means. Nectrianolins A–C ( 1 – 3 ) exhibited cytotoxic activity against both HL60 and HeLa cells.

Two new triterpene saponins, albidosides H (1) and I (2), along with the three known saponins were isolated from the barks of Acacia albida. Their structures were elucidated on the basis of extensive 1D- and 2D-NMR studies and mass spectrometry. Albidosides H (1) and I (2) were assayed for their cytotoxicity against HeLa and HL60 cells using MTT method.

Allantopyrone A is a fungal metabolite that uniquely possesses two α,β-unsaturated carbonyl moieties. We recently reported that allantopyrone A inhibited the nuclear factor-κB (NF-κB) signaling pathway induced by tumor necrosis factor (TNF)-α in human lung carcinoma A549 cells. In the present study, the mechanism by which allantopyrone A inhibits the TNF-α-induced signaling pathway was investigated in more detail. Allantopyrone A blocked extensive modifications to receptor-interacting protein 1 (RIP1) in the TNF receptor 1 (TNF-R1) complex. Allantopyrone A augmented the high-MW bands of TNF-R1, TNF receptor-associated factor 2, RIP1, the NF-κB subunit RelA and inhibitor of NF-κB kinase β in A549 cells, suggesting that it binds to and promotes the crosslinking of these proteins. The extracellular cysteine-rich domains of TNF-R1 were crosslinked by allantopyrone A more preferentially than its intracellular portion. The present results demonstrate that allantopyrone A interferes with multiple components of the TNF-R1 complex and blocks RIP1 modifications in the TNF-α-induced NF-κB signaling pathway.

A new antimalarial sterol, kaimanol (1), along with a known sterol, saringosterol (2) was isolated from the Indonesian Marine sponge, Xestospongia sp. The chemical structure of the new compound was determined on the basis of spectroscopic evidences and by comparison to those related compounds previously reported. Isolated compounds, 1 and 2 were evaluated for their antiplasmodial effect against Plasmodium falciparum 3D7 strains. Compounds 1 and 2 exhibited antiplasmodial activity with IC50 values of 359 and 0.250 nM, respectively.

Six new triterpenoid saponins, named senegalosides A–F (1–6) were isolated from the seedpods and roots of Acacia senegal (Mimosaceae). Their structures were elucidated using 1D and 2D-NMR spectroscopic analysis and mass spectrometry. Compound 1 possesses an unusual sapogenin, 3β-hydroxy-21-oxo-olean-12-en-28-oic acid (machaeric acid), and was reported here in its natural form for the first time within the genus Acacia. Senegaloside A is the first example of a machaeric-type triterpenoid glycoside in the plant kingdom. The cytotoxic effect of isolated saponins was evaluated on the H4IIE rat hepatoma cell line. As a result, compounds 1, 3–6 were not significantly cytotoxic to H4IIE cells even at 200 μM. Compound 2 was suppressed cell viability at 50–200 μM.

Cosmochlorins A (1), B (2), and C (3) were isolated from the endophytic fungus Cosmospora vilior IM2-155. The structures of 1, 2, and 3 were elucidated by a combination of extensive spectroscopic analyses, including extensive 2D NMR, HRESITOFMS, and chemical reactions. Compounds 1, 2, and 3 were evaluated for their biological activity. Compounds 1 and 2 partially restored the growth inhibition caused by hyperactivated Ca2+-signaling in mutant yeast and showed glycogen synthase kinase (GSK)-3β inhibition activity at IC50 values of 62.5 and 60.6 μM, respectively. Further, compound 2 significantly increased osteoclast formation by more than 1.5-fold in RAW264.7 cells compared to receptor activator of nuclear factor-κB ligand (RANKL) alone.

The genome of Aspergillus oryzae encodes a hypothetical protein (annotated as AO090701000884), designated AoFaeD, which exhibits sequence identity to feruloyl esterases from Aspergillus clavatus, Coprinopsis cinerea, Neurospora crassa , and Pseudomonas fluorescens subsp. cellulosa . In this study, we cloned the AofaeD coding sequence into Pichia pastoris and demonstrated heterologous expression and secretion of active recombinant enzyme (rAoFaeD) as a 30-kDa protein in the culture medium. Purified rAoFaeD exhibited optimal activity at pH 7.0 and at 45 °C, but was stable at a pH range of 7.0–10.0 and a temperature of 40 °C. While wild-type rAoFaeD failed to cleave the methyl esters of ferulic acid (MFA), p -coumaric acid (M p CA), caffeic acid (MCA), and sinapic acid (MSA) and ethyl ester of ferulic acid (EFA), enzyme exhibited esterase activity when wheat arabinoxylan was used as a substrate, and this reaction was enhanced synergistically by the addition of xylanase. Notably, rAoFaeD variants in which one of the tryptophan residues (Trp142 or Trp144) located adjacent to the putative catalytic serine residue at position 143 (Ser143) was replaced with phenylalanine and tyrosine (W142F and W144Y), respectively, exhibited hydrolytic activity toward MFA, M p CA, MCA, MSA, and EFA. Moreover, these variants exhibited enhanced production of ferulic acid from wheat arabinoxylan compared to the wild-type protein, and the activity of these proteins was enhanced by the addition of xylanase.

Phytochemical investigation of the roots of Albizia chevalieri led to the isolation of two new 5-deoxyflavan-3,4-diol glucosides from roots of A. chevalieri, Chevalieriflavanosides A and B. Their structures were established by 2D NMR techniques, UV, IR, CD, and mass spectrometry. Cytotoxicity of the two compounds was evaluated against acute promyelocytic leukemia HL60 cells. The antibacterial activities of 1 and 2 also were evaluated against Pseudomonas aeruginosa and Staphylococcus aureus using the agar diffusion test. Copyright © 2016 John Wiley & Sons, Ltd.

Three novel benzenediol lactone derivatives, 7-hydroxy-14-de-O-methyl-lasiodiplodin (2), ethyl-3,5-dihydroxy-7-(6,8-dihydroxynonyl)benzoate (4), and 7-ethylbenzoate-7-heptanoic acid (5), together with known compounds, 14-de-O-methyl-lasiodiplodin (1) and ethyl-3,5-dihydroxy-7-(8-hydroxynonyl)benzoate (3), were isolated from the culture of Fusarium solani T-13, a fungus from a dead branch. Their structures were elucidated using 1D- and 2D-NMR spectroscopy. Compounds 1, 3 and 4 showed doughnut like phenotypes of growth restoring activity against the mutant yeast (zds1ο erg3ο pdr1/3ο) at the concentration of 0.16, 0.63 and 1.25 μg/spot, respectively.

Keap1-Nrf2 system is known as a sensor of electrophilic compounds, and protects cells from oxidative stress through induction of various antioxidant enzymes. We found by proteomic analysis that allantopyrone A, a metabolite isolated from an endophytic fungus, upregulates the expression of proteins that are regulated by the transcription factor Nrf2. Indeed, allantopyrone A increased the antioxidant enzyme heme oxygenase-1 in PC12 cells. Moreover, it induced localization of Nrf2 in the nucleus. Affinity purification of allantopyrone A-binding protein showed that this compound could bind directly to Keap1. Allantopyrone A suppressed intracellular reactive oxygen species level and cell death induced by H2O2 in PC12 cells. These results indicate that allantopyrone A protects PC12 cells from oxidative stress-induced cell death through direct binding with Keap1 and activation of the Keap1-Nrf2 pathway.

One new polyketide derivative, heptacyclosordarianone (1), together with 2 known compounds, heptacyclosordariolone (2) and sordariol (3), were isolated from an extract of Sordaria sp. AM-71, an endophytic fungus inhabiting Garcinia polyantha. The structures of these compounds were elucidated based on intensive nuclear magnetic resonance spectroscopy and high-resolution mass spectrometry and by comparison with literature data. All compounds were assayed for their antimicrobial and cytotoxic properties. Only compound 2 showed antimicrobial activities against Bacillus subtilis DSMZ 704 , Pseudomonas agarici DSMZ 11810, and Micrococcus luteus DSMZ 1605 with minimum inhibitory concentration values of 9.3, 15.5, and 16.9 µg/mL respectively. None of the isolated compounds showed a significant cytotoxic property.