Antimicrobial and cytotoxic activities of marine sponge-derived fungal extracts isolated from Dactylospongia sp.

This study is the first report on the antimicrobial and cytotoxic activities of fungal extracts isolated from marinesponge Dactylospongia sp., which is collected from Mandeh Island, West Sumatra, Indonesia. The isolation of fungalwas conducted using dilution method with Sabouraud Dextrose Agar + chloramphenicol (0.05%) as a medium. Thepure isolated fungal was cultivated on rice medium at temperature 25°C–27°C and then extracted using ethyl acetatesolvent. The ethyl acetate extract of each isolated fungal was tested for antimicrobial and cytotoxic activities. Ninefungal strains have been isolated from this sponge. Two ethyl acetate extracts of fungal strains (Dc03 and Dc04) werecategorized as having strong inhibition against the growth of Staphylococcus aureus, Escherichia coli, methicillinresistant S. aureus, and multidrug-resistant Pseudomonas aeruginosa in a concentration of 5% with zone inhibitionin range of 12.31 ± 0.54–16.14 ± 0.75 mm. The cytotoxic activity screening of the ethyl acetate extracts of fungalstrains was done by using the brine shrimp lethality test. Four fungal strains had LC50 below 80 µg/ml (Dc03, Dc04,Dc05, and Dc08) and were further tested with MTT (3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide)assay on T47D cell line. These selected fungi were identified molecularly as Cladosporium halotolerans MN859971,Penicillium citrinum MN859968, Aspergillus versicolor MN859970, and Aspergillus sydowii MN859969, respectively.The results suggest that these fungal strains are quite rich in the production of bioactive compounds that are veryeffective as antibacterial and cytotoxic agents

A new fumagillin compound from marine-derived Aspergillus fumigatus MDCW-15 / 中草药

Objective: To study the secondary metabolites of marine-derived Aspergillus fumigatus MDCW-15. Methods: The secondary metabolites were isolated and purified by column chromatography over silica gel. And their structures were identified by the spectroscopic analysis of NMR and MS. The antifungal bioactivities were assayed by paper diffusion. Results: A new fumagillin compound 1 and a known compound 2 were isolated from the fermentation broth of marine-derived Aspergillus fumigatus MDCW-15. The antifungal bioactivities were assayed by paper diffusion. Compounds 1 and 2 showed antifungal activity against Candida albicans with an equal MIC value of 32.0 μg/mL. Conclusion: Compound 1 is a new compound named 2'-cis- fumagiringillin. Compounds 1 and 2 exhibit antifungal activities.

Penicacids E-G, three new mycophenolic acid derivatives from the marine-derived fungus Penicillium parvum HDN17-478 / 中国天然药物

Three new mycophenolic acid derivatives, penicacids E-G (1-3), together with three known analogues, mycophenolic acid (4), 4'-hydroxy-mycophenolic acid (5) and mycophenolic methyl ester (6), were isolated from a marine-derived fungus Penicillium parvum HDN17-478 from a South China Sea marine sediment sample. The structures of compounds 1-3 were elucidated by HRMS, NMR, and Mosher's method. Among them, compounds 1 and 2 were the first examples of mycophenolic acid analogs with a double bond at C-3'/C-4' position. The cytotoxicity of 1-6 was evaluated against the HCT-116, BEL-7402, MGC-803, SH-SY5Y, HO-8910 and HL-60 cell lines, and compounds 4 and 6 showed potent cytotoxicity with IC

In vitro cytotoxic activity assay of bacteria extract derived marine sponge Haliclona fascigera toward Hela, WiDr, T47D, and Vero cell line

Symbiotic association between marine sponge and microorganism was a promising chance in the discovery of leadcompound of anticancer. This association was probably concluded that symbiotic microorganism would contain thesame secondary metabolites with the host. In this continuation research, we had isolated symbiotic bacteria from amarine sponge and tested for cytotoxic activity. Twenty-six isolates of bacteria derived from marine sponge Haliclonafascigera were isolated from Setan Island, West Sumatra, Indonesia. Screening of cytotoxic activity by BSLT methodand MTT assay was conducted toward 21 ethyl acetate extracts of symbiotic bacteria with weight >50 mg. Onebacterial extract with code H2N was very toxic according to BSLT test, while 18 isolates were toxic with LC50 rangingfrom 31.17 to 283.38 ppm. All of the bacterial extracts did not show a good percentage of viability (>50%) againstHela, WiDr, T47D, dan Vero cell line in MTT assay. However, bacterial extract with code H2N have shown potentialcytotoxic compared to other extracts. As per the phytochemical study, this extract probably contained terpenoid group.Based on biochemical examination this bacterium, H2N, was identified as Bacillus sp.3.

A new glycosyl ester isolated from marine-derived Penicillium sp / 中草药

Objective: To isolate and identify the active natural products of marine-derived Penicillium sp. H1. Methods: The isolations and purifications of secondary metabolites were performed by means of column chromatography over silica gel. And their structures were elucidated through the spectroscopic analysis of MS, NMR, and specific rotations. The bioactivities were assayed by paper diffusion. Results: From the fermentation broth of marine-derived Penicillium sp. H1, five compounds were isolated and identified as xylarinonericin E (1), fumitremorgin C (2), verruculogen (3), penicillide (4), and pyripyropene A (5), respectively. Compound 1 showed moderate antifungal activity against Fusarium oxysporum f. sp. Cubense with MIC value of 32.0 μmol/mL. Compound 2 displayed antibacterial activity against Staphylococcus aureus with MIC value of 64.0 μg/mL. Conclusion: Compound 1 is a new compound. Compounds 1 and 2 exhibit moderate antimicrobial activities.

Secondary metabolites from marine-derived fungus Aspergillus sp. SCS-KFD66 / 中草药

Objective: To study the secondary metabolites from the fermentation broth of marine-derived fungus Aspergillus sp. SCS-KFD66. Methods: The constituents were isolated and purified by silica gel, Sephadex LH-20 column chromatography and HPLC methods. The structures of the compounds were identified by spectral data analysis. The DPPH radical scavenging, acetylcholinesterase and α-glucosidase inhibitory activities of compounds were evaluated by DPPH method, Ellman colorimetric method and PNPG method, respectively. The inhibitory effect and the minimal inhibitory concentration (MIC) of compounds on Escherichia coli, Staphylococcus aureus, Bacillus subtilis, and Listeria monocytogenes were tested using 96-well microtiter plates method. Results: A total of 13 compounds were isolated from the fermentation broth of marine-derived fungus Aspergillus sp. SCS-KFD66 and identified as 3-epi-trans-4-hydroxylinalool 3,6-oxide (1), trans-4-hydroxylinalool 3,6-oxide (2), aloe emodin (3), emodin (4), citreorosein (5), protocatechualdehyde (6), 2,5-dihydroxybenzaldehyde (7), methyl 4-hydroxyphenylacetate (8), 4-hydroxybenzoic acid (9), 4-hydroxyphenylacetic acid (10), 2-(4-hydroxyphenyl) ethanol (11), (E)-p-hydroxycinnamic acid (12) and (E)-ferulic acid (13), respectively. Compounds 1, 6, 7 and 9-13 showed DPPH radical scavenging activities; Compounds 4 and 6 showed acetylcholinesterase inhibitory activity; Compounds 6 and 7 exhibited inhibitory activity against α-glucosidase. Compound 4 has inhibitory activity against S. aureus and B. subtilis with the MIC values of 16 μg/mL and 64 μg/mL, respectively. Compound 6 showed inhibitory activity against E. coli, B. subtilis and L. monocytogenes, with MIC values of 64, 128 and 128 μg/mL, respectively. Conclusion: Twelve known compounds and one new compound were isolated. Compound 1 is a new compound named as aspergfuranol.

New polyketides from chemical mutant strain of sponge-associated fungus emericella variecolor xsa-07-2 / 中国药学杂志

OBJECTIVE: To study the new bioactive secondary metabolites of diethyl sulfate chemical mutant strain of sponge-associated fungus Emericella variecolor XSA-07-2. METHODS: Diethyl sulfate was used to make chemical mutagenesis of strain XSA-07-2, and one mutant strain M8 was chosen for large-scale fermentation to generate new secondary metabolites. The compounds were isolated and purified by chromatography on silica gel and ODS reversed-phase column and semi-preparative HPLC techniques. And their structures were identified by their physicochemical properties and NMR, MS data analysis. RESULTS: Three new polyketides 1-3 were isolated from the extract of the solid fermentation culture of mutant strain M8. Compound 3 showed moderated antioxidant activity with IC50 of (13.58±0.14) μg·mL-1 by DPPH assay. CONCLUSION: Diethyl sulfate chemical mutagenesis can stimulate sponge-associated fungus Emericella variecolor XSA-07-2 mutant strain M8 to produce new antioxidant polyketides.

A new polyketide from marine-derived Streptomyces sp.MDW-06 / 中国中药杂志

To isolate and identify secondary metabolites of marine-derived Streptomyces sp.MDW-06,the isolations and purifications of compounds were performed by means of column chromatography over silica gel. And their structures were elucidated through the spectroscopic analysis of MS,NMR and specific rotations. The bioactivities were assayed by paper diffusion and DPPH method. From the fermentation broth of marine-derived Streptomyces sp.MDW-06,five compounds( 1-5) were isolated and identified as streptopentanoic acid( 1),germicidin A( 2),germicidin B( 3),isogermicidin A( 4),isogermicidin B( 5) and oxohygrolidin( 6),respectively. Compound 1 is a new compound. Compound 1 shows DPPH radical scavenging activity with 36. 4% at 100 mg·L~(-1).

Alkaloids from the Marine-Derived Fungus Aspergillus fumigatus YK-7 and Their Antitumor Activities / 中国药学杂志

OBJECTIVE: To study the secondary metabolites of marine-derived fungus Aspergillus fumigatus YK-7. METHODS: The compounds were isolated by several column chromatographic techniques, including silica gel, ODS, Sephadex LH-20 column chromatography, and HPLC, and their structures were identified on the basis of physicochemical properties and spectroscopic analysis. Trypan blue and MTT methods were applied for determining the effects of the compounds on proliferation of cancer cells in vitro. RESULTS: Ten compounds were obtained, and their structures were identified as pseurotin A (1), pseurotin A1(2), 14-norpseurotin A (3), FD-838 (4), demethoxyfumitremorgin C (5), 9β-hydroxyverruculogen TR-2 (6), 6-methoxyspirotryprostatin B (7), spiro[5H, 10H-dipyrrolo[1, 2-a:1', 2'-d]pyrazine-2-(3H), 2'-[2H]indole]-3', 5, 10(1'H)-trione (8), terezine D (9), and 14-hydroxyterezine D (10). CONCLUSION: Compounds 3, 6, 7, 9, and 10 are isolated from marine-derived fungus Aspergillus fumigatus for the first time. Compounds 1-4 exhibite moderate antiproliferative activity against selected cancer cell lines in vitro.

Potential of four marine-derived fungi extracts as anti-proliferative and cell death-inducing agents in seven human cancer cell lines

OBJECTIVE@#To evaluate the in vitro anticancer activity of crude ethyl acetate extracts of the culture of four marine-derived fungi Aspergillus similanensis KUFA 0013 (E1), Neosartorya paulistensis KUFC 7897 (E2), Neosartorya siamensis KUFA 0017 (E4) and Talaromyces trachyspermus KUFC 0021 (E3) on a panel of seven human cancer cell lines.@*METHODS@#Effects on cell proliferation, induction of DNA damage and cell death were assessed by MTT and clonogenic assays, comet assay and nuclear condensation assay, respectively.@*RESULTS@#The proliferation of HepG2, HCT116 and A375 cells decreased after incubation with the extracts E2 and E4. The anti-proliferative effect was confirmed by morphologic alterations and by clonogenic assay. Both extracts also induced cell death in HepG2 and HCT116 cells. Doxorubicin was used as a positive control and showed in vitro anticancer activity.@*CONCLUSIONS@#This study demonstrated, for the first time, that extracts of Neosartorya paulistensis and Neosartorya siamensis have selective anti-proliferative and cell death activities in HepG2, HCT16 and A375 cells. The bioactivity of these extracts suggests a potential for biotechnological applications and substantiates that both should be further considered for the elucidation of the molecular targets and signal transduction pathways involved.

Chromopeptide A, a highly cytotoxic depsipeptide from the marine sediment-derived bacterium Chromobacterium sp. HS-13-94

A bicyclic depsipeptide, chromopeptide A (1), was isolated from a deep-sea-derived bacterium Chromobacterium sp. HS-13-94. Its structure was determined by extensive spectroscopic analysis and by comparison with a related known compound. The absolute configuration of chromopeptide A was established by X-ray diffraction analysis employing graphite monochromated Mo K α radiation (λ=0.71073 Å) with small Flack parameter 0.03. Chromopeptide A suppressed the proliferation of HL-60, K-562, and Ramos cells with average IC50 values of 7.7, 7.0, and 16.5 nmol/L, respectively.

Penidioxolanes A and B, 1,3-Dioxolane Containing Azaphilone Derivatives from Marine-derived Penicillium sp. KCB12C078

Two new azaphilone derivatives containing 1,3-dioxolane moiety, penidioxolanes A (1) and B (2), were isolated from marine-derived fungus Penicillium sp. KCB12C078, together with four known compounds (3-6) by chemical investigation. Compounds 1 - 6 were isolated by combination of silica gel, ODS column chromatography and preparative HPLC. Their structures were determined by analysis of spectroscopic data including 1D-, 2D-NMR, and MS techniques. The isolates were evaluated against cancer cell growth inhibition effects and antimicrobial activity.

Potential of four marine-derived fungi extracts as anti-proliferative and cell death-inducing agents in seven human cancer cell lines

Objective: To evaluate the in vitro anticancer activity of crude ethyl acetate extracts of the culture of four marine-derived fungi Aspergillus similanensis KUFA 0013 (E1), Neosartorya paulistensis KUFC 7897 (E2), Neosartorya siamensis KUFA 0017 (E4) and Talaromyces trachyspermus KUFC 0021 (E3) on a panel of seven human cancer cell lines. Methods: Effects on cell proliferation, induction of DNA damage and cell death were assessed by MTT and clonogenic assays, comet assay and nuclear condensation assay, respectively. Results: The proliferation of HepG2, HCT116 and A375 cells decreased after incubation with the extracts E2 and E4. The anti-proliferative effect was confirmed by morphologic alterations and by clonogenic assay. Both extracts also induced cell death in HepG2 and HCT116 cells. Doxorubicin was used as a positive control and showed in vitro anticancer activity. Conclusions: This study demonstrated, for the first time, that extracts of Neosartorya paulistensis and Neosartorya siamensis have selective anti-proliferative and cell death activities in HepG2, HCT16 and A375 cells. The bioactivity of these extracts suggests a potential for biotechnological applications and substantiates that both should be further considered for the elucidation of the molecular targets and signal transduction pathways involved.

Graphiumins I and J, New Thiodiketopiperazines from the Marine-derived Fungus Graphium sp. OPMF00224

Two new thiodiketopiperazines (TDKPs), designated graphiumins I (1) and J (2), were isolated from the culture broth of the marine-derived fungus Graphium sp. OPMF00224 by solvent extraction, silica gel column chromatography, and HPLC. Their absolute structures were elucidated by spectroscopic analyses (1D and 2D NMR data, ROESY correlations, and CD data) and chemical methods. They were found to be structurally rare TDKPs with a phenylalanine-derived indolin substructure. Compounds 1 and 2 inhibited yellow pigment production by methicillin-resistant Staphylococcus aureus (MRSA) with IC50 values of 63.5 and 76.5 microg/ml, respectively, without inhibiting its growth, even at 250 microg/ml.

Effect of various media and supplements on laccase activity and its application in dyes decolorization

Aims: Marine-derived fungi are a potential for the search of new compounds with relevant features. Among these, the ligninolytic enzymes have potential applications in a large number of fields, including the environmental and industrial sectors. This work aimed to evaluate the laccase activity of the marine-derived fungus Alternaria alternata, under various cultivation conditions and its application in synthetic dyes decolorization. Methodology and results: Wheat bran prepared with 40 mL sea water proportion was the most suitable substrate for laccase production (114.06±2.24 U/mL) by A. alternata, after 14 days of incubation in submerged fermentation. Laccase production in static cultivation was superior to that in agitated cultures. The simple Boyd and Kohlmeyer medium with supplementation of 2 mM CuSO4·5H2O on day 6, at an incubation period of 14 days and incubation temperature of 28±2°C under static conditions, yielded amounts of laccase (36.13±0.34 U/mL) less than that obtained with submerged fermentation of wheat bran as unique substrate. Furthermore, A. alternata has high decolorization capability toward azo dyes in the absence of redox mediators, 75.47% of the reactive black at 0.01% concentration, was removed after 30 days of incubation. Also has good ability to decolorize the triphenylmethane dye crystal violet, at 0.01% concentration, about 69.35% of the dye was removed after 30 days. Conclusion, significance and impact of study: These unusual properties demonstrate that the marine-derived fungus Alternaria alternata has potentials in specific industrial or environmental applications.

Evaluating methods for the isolation of marine-derived fungal strains and production of bioactive secondary metabolites

In the present investigation we evaluate methods for the isolation and growth of marine-derived fungal strains in artificial media for the production of secondary metabolites. Inoculation of marine macroorganisms fragments in Petri dishes proved to be the most convenient procedure for the isolation of the largest number of strains. Among the growth media used, 3% malt extract showed the best result for strains isolation and growth, and yielded the largest number of strains from marine macroorganisms. The percentage of strains isolated using each of the growth media which yielded cytotoxic and/or antibiotic extracts was in the range of 23-35%, regardless of the growth media used. Further investigation of extracts obtained from different marine-derived fungal strains yielded several bioactive secondary metabolites, among which (E)-4-methoxy-5-(3-methoxybut-1-enyl)-6-methyl-2H-pyran-2-one is a new metabolite isolated from the Penicillium paxilli strain Ma(G)K.

Study on the secondary metabolites of marine-derived fungus Penicillium sacculum / 中国药学杂志

OBJECTIVE: To study the secondary metabolites of marine-derived fungus Penicillium sacculum. METHODS: The compounds were separated by several column chromatographic techniques, and their structures were elucidated by means of physico-chemical properties and spectral data. Trypan blue exclusion assay and MTT method were used to test the anti-tumor activities. RESULTS: Ten compounds were isolated from the fermentation broth and mycelium. Their structures were identified as(R)-1, 3-di-hydro-6-methoxy-3-methylisobenzofuran-4, 5-diol(curvulol)(1), griseofulvin(2), 1, 6-dihydroxy-3-methoxy-8-methylxanthone(3), 1-hydroxy-3-methylxanthone(4), 1, 3, 6-trihydroxy-8-methylxanthone(5), 1, 8-dihydroxy-3-methylanthraquinon(6), 1, 3, 5-trihy-droxy-7-methylanthraquinone(7), citreorosein(8), adenine(9) and thymine(10). Among them, compounds 1 and 3 showed respectively potent and moderate growth inhibitory activities against human leukemia HL-60 cells. CONCLUSION: Compounds 1 and 4 were isolated for the first time from the genus of marine-derived fungus. The 1H-NMR and 13C-NMR signals of compound 1 were assigned for the first time. Compounds 1 and 3 have anti-tumor activities. Copyright 2012 by the Chinese Pharmaceutical Association.

A new approach for exploiting microbial new strain resources for drug screening / 国际药学研究杂志

Generally absolute majority of wild-type microbial strains do not produce bioactive metabolites, resulting in large numbers of so-called 'useless strains' stocked or destroyed. These strains, however, would become a great source of bioactive meabolites if their secondary metabolism could be altered to produce diverse metabolites. We have therefore undertaken a research work on exploiting microbial new strain resources for drug screening by altering secondary metabolism of the 'useless strains' to discover bioactive metabolites. A considerable progress with expectant advantage desired has been made in the studies on marine-derived actinomycetic and fungal strains. This paper summarizes our research results including several new developments in brief.

Screening for bioactive mutants with antitumor activity from an actinomycetic wild-type strain without antitumor activity by antibiotic-resistant mutation technique and by coupled with chemical mutagen-induced mutation / 军事医学科学院院刊

Objective To obtain antibiotic-resistant mutants producing metabolites with antitumor activity from wild-type actinomycete strains without antitumor activity. Methods An actinomycete strain L35-1 was used as an initial strain for obtaining antibiotic-resistant mutants, which is a marine-derived wild-type strain without antitumor activity with an inhibition rate of 2.8% at the 1000 μg/ml of high sample concentration on K562 cells. The antibiotic-resistant mutants both from auto-mutagenesis and chemical mutagen-induced mutagenesis were selected by single colony isolation on antibiotic-containing plates according to the method for obtaining drug-resistant mutants in ribosome engineering. The antitumor activity was assayed by the MTT method using K562 cells for the mutants with aqueous acetone extracts of the whole broth of their fermentation.Results A total of 114 neomycin-resistant (ner) and 68 streptomycin-resistant (str) mutants, all from auto-mutagenesis, was obtained on drug-containing plates. Among them, the 7 ner and 3 str mutants appeared to be bioactive with an inhibition rate above 20% at the 100 μg/ml sample concentration on K562 cells. On the other hand, 41 str and 32 ner mutants from DES-induced mutagenesis and 46 ner mutants from NTG-induced mutagenesis were obtained by mutagen-induced mutation coupled with the single colony isolation on antibiotic-containing plates, among which, one str mutant from DES-induced mutagenesis and one ner mutant from NTG-induced mutagenesis were bioactive with an inhibition rate over 20% at the 100 μg/ml sample concentration on K562 cells. Conclusions The present result has revealed that the wild-type actinomycete strains without bioactivity might become a great source initial strains to obtain bioactive mutants by drug-resistant mutation technique.

Tyrosinase inhibition activity of Botrytis sp.metabolites a-pyrone / 中国海洋药物

Anti-tyrosinase assay guided isolation of marine-derived fungus Botrytis sp.led to separation of three ?-pyrone compounds.The structures of these compounds were elucidated by various physicochemical analytical methods.The bioactivities of these ?-pyrone derivatives were evaluated in terms of anti-tyrosinase activity.By correlating the bioactivities of these compounds with their structures,it concluded that the ?-pyrone with pentyl group gives highest anti-tyrosinase activity.