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Studies on Halotolerant Fungal Strains: Isolation, Culture and Secondary Metabolites

Author: WangWenLiang
Tutor: ZhuWeiMing;GuQianQun
School: Ocean University of China
Course: Medicinal Chemistry
Keywords: Salt-tolerant microorganisms Genius strains Secondary metabolites Anti-tumor Active track separation
CLC: R284
Type: PhD thesis
Year: 2008
Downloads: 642
Quote: 3
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Salt-tolerant microbes to survive and develop in extreme environments, in this particular environment may activate a certain class of microorganisms silence genes, resulting in the special structure of the protein, thereby inducing a unique biological pathway, making secondary metabolite structure novel, and variety. Therefore, the salt-tolerant microorganisms is an important source of the new structure of the active compounds. In order to find a the genius of strains the (Talented strain) and novel structure of the active compound with a salt-tolerant microorganisms, this paper uses a chemical and biological activity of the combination of an integrated screening method, to carry out the research work of the production of anti-tumor activity of metabolites halophilic Microorganism . The contents include: salt-tolerant microorganisms separation and anti-tumor activity screening; preliminary evaluation of the active strains, strains obtained genius and tracking of the anti-tumor activity component separation; structural analysis of the monomer compounds; monomer compounds the preliminary evaluation of anti-tumor activity. Shrimp bio-lethal method K562 cells by flow cytometry screening model, antioxidant and antibacterial test screening model, inhibition of cell cycle, apoptosis induction, combined with chemical screening, 19 were collected from Inner Mongolia Jilantai the saltworks and high-salt environment of the Qinghai Lake samples (13 mud samples, six water samples) were isolated 174 salt-tolerant microorganisms screened three active bacteria, a genius strains. Means of thin-layer chromatography, column chromatography on silica gel, Sephadex LH-20 column chromatography, reversed-phase high performance liquid chromatography separation combined with physical and chemical constants and by analyzing the spectral data clarify the discoloration from the salt-tolerant Aspergillus Aspergillus variecolor B-17 were isolated 49 the structure of the compound of: variecolortide AC (1-3), variecolorins AN (4-17), neoechinulin A (18), preechinulin (19), neoechinulin B (20), (21), dihydroxyisoechinulin (22), isoechinulin A (23), tardioxopiperazine A (24), tardioxopiperazine B (25), isoechinulin B (26), echinulin (27), Alkaloid E-7 (28), cryptoechinuline G (29), variecolorquinones B (30), variecolorquinones A ( 31), (2S) -2,3-dihydroxypropyl-1 ,6,8-trihydroxy-3-methyl-9 ,10-dioxoanthracene-2-carboxylate (32), emodin (33), physcion (34), questin ( 35), questinol (36), catenarin (38), erythroglaucin (39), rubrocristin (40), eurotinone (41), 2-methyleurotinone (42), flavoglaucin (43), aspergin (44), dihydroauroglaucin (45), 2 - (1,1-dimethyl-2-propen-1-yl)-1H-Indole-3-carboxaldehyde (46), diisobutyl phthalate (47), butyrolactone I (48), halobutyrolactone A (49); from another 27 structure of the compound isolated strains halophilic Microorganism THW-18: halotolerantcerebroside AC (50-52), cerebroside C (53), cerebroside D (54), (22E, 24R)-3β, 5α-Dihydroxy-23 -methylergosta-7 ,22-dien-6-one (55), (22E, 24R)-3β, 5α-Dihydroxy-ergosta-7 ,22-dien-6-one (56), (22E, 24R)-3β , 5α, 9α-trihydroxyergosta-7 ,22-diene-6-one (57), (22E, 24R)-23-Methylergosta-7 ,22-diene-3β, 5α, 9α-triol (58), cerevisterol (59 ), 3β, 5α-dihydroxy-6β-methnoxyergosta-7, 22 - diene (60), ergosterol peroxide (61), ergosterol (62), demethylincisterol A3 (63), haloacetylaranotin A (64), bisdethiodi (methylthio) acetylaranotin ( 65), acetylaranotin (66), alterperylenol (67), dihydro-alterperylenol (68), altertoxin I (69), bis (2-ethylhexyl) phthalate (70), N-acetyltyramine (71), cyclo-(Tyr-Pro ) (72), (15Z)-octadecenoic acid (73), (13Z)-hexadecenoic acid (74), benzoic acid (75), guanosine (76). Alkaloids 1-17, quinone compounds 30-31, butyrolactone compounds 49, cerebroside compounds 50-52, sterols 56 and sulfur-containing cyclic peptide compounds 64 are new compounds . Flow cytometry combined morphological detection, MTT and SRB assay, in vitro anti-tumor activity of the share of the new structure of the compounds were carried out a preliminary evaluation. The results showed that the new compounds 1-3 on K562 cell proliferation inhibitory activity of the weak; new compounds 4-17 P388, A-549, HL-60 and BEL-7402 cells have weak cytotoxic activity; compound 30 on a variety of human cancer cells and mammalian cancer cells have stronger cell proliferation inhibitory activity; 31 on human lung cancer A-549 cells, a new compound having strong cytotoxic activity; new sterols Compound 56 on HL-60 cells The proliferation inhibiting activity. Radical scavenging test, a New Compound 17 showed a strong free radical scavenging activity; radical scavenging activity of the new compounds 1-14 and 31 show the moderate intensity; radical scavenging activity of the new compounds 64 shows weak; known Compound 41 -45 show a strong radical scavenging activity; the compound 18,20,22,23,26,28,29,32-40 radical scavenging activity of moderate intensity; 67-69 show weak radical scavenging activity. In summary, this article by the activity screening from 174 salt-tolerant microorganisms three active bacteria, of which a \76 of the compound, wherein the new compounds 25. First discovered three novel structure of the anthraquinone and diketopiperazine-based compound polymer backbone compounds, other new compounds, halogen-containing compound also contains 3 molecule. Biological tests showed that these new compounds have a certain degree of anti-tumor activity in vitro. The above study microbial resources and structure of a novel screening compounds for anti-cancer agents, but also provides a guideline for the design and synthesis of new halogenated drugs.

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