| The active compounds were isolated and identified, which were from Piptoporusbetulinus, an endophytic bacterium of Gingko and endophytic actinomycetes ofGanoderma. Study the bioactive of some metabolites and others. The result weresummarized as following:Firstly, column chromatograph with different medium was resorted to isolate andpurify the secondary metabolites of the fruiting body of Pipoporus betulinus. NMRand other spectroscopy were used to identify the structures of compounds.15compounds were determined from24compounds which were isolated: Stigmast-5-en-3-ol, Stigmast-7-en–3-ol, Stigmast-9-en-3-ol, Erogosta-7-en-3-ol, Erogosta-7-dien-3-ol, Erogosta-7-en,16-hydroxy-3-oxolanosta-7,9(11),24-trien-21-oic-acid, Botulin,16-hydroxy-3-acetoxylanosta-7,9(11)-dien-21-acid,Fagarsterol-20-en,3-oxolanosta-8,24-dien-21-oic acid,16-dydroxy-3-oxolanosta-7,9(24)-trien-21-oic acid,16-dydroxy-lanosta-7,9(24)–trien-21-oicacid,3–oxolanosta-7,9(24)-trien-21-oic acid and Ergosta-7,22–dien–3-one.All above compounds were all found in the Pipoporous betulinus firstly.Secondly, the antitumor activity of extracts against NCI-H460and SGC-7901was performed by using Alamar Blue method. The results showed that the antitumoractivity of petroleum ether extract was strong against NCI-H460and SGC-7901celllines; however, ethyl acetate and methanol extracts have different effect. Theinhibition value of methanol extracts was lower than petroleum on the NCI-H460cellline and the ethyl acetate was not having activity almostly.Thirdly, optimize the conditions of fermentation for mycelia that isolated fromthe Pipoporus betulinus and the index were the concentration of biomass,polysaccharide and triterpene. One primary condition was got by the study: basal medium, initial pH=6.5, educate the strain at the temperature of30℃for140h. Cornstarch was chosed as carbon source and the beef extract was used as nitrogen sourcefollowing the designing. Meanwhile, the phenol has no effect by the nutri stress butthe triterpene was not.Fourthly, an endophytic bacterium, designated strain Bacillus amyloliquefaciensCGMCC5569was isolated from Chinese medicinal Ginkgo biloba collected fromXuzhou, China. Both the filtrate and the ethyl acetate extract of strain CGMCC5569showed growth inhibition activity against the sapstain fungi Lasiodiplodiarubropurpurea, L. crassispora and L. theobromae obviously (>65%) based on thecomparison of the lengths of zones on the petri dish. Four compounds were isolatedand identified by NMR, IR and MS. There were Nystatin,6-hydroxypropyl-2,4-amidelactone,6-hydroxylbutyl-2,4-amide lactone,6-ethoxyl--2,4-amide lactone, biuret.Meanwhile, the part H was isolated by the HSCCC and identified. A highest fractionwas obtained following activity guide and identified by LC-MS. It was a mixture ofSurfactin C12~C15，Fengycin A C14~C17，Fengycin B C17，Bacillomycin L C14，Bacillomycin L C15，Bacillomycin D C14，Bacillomycin D C15. It showed stronggrowth inhibition activity in vitro against the sapstain. It showed strong growthinhibition activity in vitro against the L. rubropurpurea, L. crassispora and L.theobromae by about70.22%,69.53%and78.76%, respectively. The stronganti-sapstain fungus activity indicated that the endophytic B. amyloliquefaciensCGMCC5569and its bioactive components might provide an alternative bio-resourcefor the bio-control of sapstain.Fifthly, an endophytic Streptomyces recifensis CGMCC5228has been isolatedin our laboratory from Ganoderma and showed high levels of inhibits tumor growth.The extract of chloroform showed highest active of inhibited the growth of tumor cellline SGC-7901and NCI-H460. Compound LZ-B, LZ-H and LZ-E were isoloated andidentified from the extract of chloroform using active guide. The compound of LZ-Bwas identified as a new compound named2-aminomethyl-actinomycin D and theLZ-H was named actinomycin D. Another compound was identified as a known named Olanzapine. To the best of our knowledge, it was the first report that theolanzapine was isolated from natural. The antitumor activity of compound LZ-Bagainst NCI-H460and SGC-7901was performed by using Alamar Blue method. Theresults showed that the antitumor activity of LZ-B was strong against SGC-7901cellline. The inhibition value of SGC-7901was higher than the NCI-H460. The value was86.37%for SGC-7901when the concentration of LZ-B was80ng/mL, but theNCI-H460was only73.36%。The interaction of2-aminomethyl-actinomycin D withBovine Hemoglbin (BHb) awas investigated by UV/vis absorption and fluorescencespectra techniques. The results showed that the interaction of2-aminomethyl-actinomycin D with BHb was static quenching. And the proteinstrcutre was changed to close-knit by the interaction.The condition of HPLC for the2-aminomethyl-actinomycin D was got asfollowing: gradient elute with methonal and water (45%methonal) and the detectionwavelength was254nm. The carbon and nitrogen source was optimized following themethod of HPLC and the index was the concentration of2-aminomethyl-actinomycinD. The results showed that the table sugar was the best carbon source and the nitrogensource was beef extract. Response surface method was used in the optimization thecondition of the fermentation and got a better condition by this method. Theconcentration of2-aminomethyl-actinomycin D was reached at0.0447mg/mL usingfollowing condition:: pH=8.0, temperature27℃, the rotate speed=50r/min and thetime was6.5days, respectively. |
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