| The harsh living conditions like high radiation or low temperature force microorganisms in the polar region to create many physical or chemical protection mechanisms.The enzyme reaction system and the biosynthetic pathway of polar microorganism differ a lot from ordinary terrestrial microorganism.So polar microorganism can produce compounds with novel structure and unique activity.As a result,the polar region is an important repository of microorganism,and is also one of the most promising area of developing.new drugs.In our precious work,the cytotoxic activity screening of a number of strains from the Antarctica was adopted.The strain Penicillium sp.S-3-88 showed outstanding cytotoxic activity against six kinds of tumor cells(H446,A549,SW-1990,MCF-7,HeLa and SGC-7901).This indicated that Penicillium sp.S-3-88 could produce active chemical substances that may show strong inhibitory effect on the growth of tumor cells.In present work,the effects of fermentation medium,temperature,rotation speed and cultural time on antitumor activity and secondary metabolites amount were analyzed firstly.The appropriate fermentation condition was: potato-dextrose-broth medium,NaCl 25 g·L-1,20 ℃,180 r·min-1,for 14 d.After the fermentation conditions were optimized,the secondary metabolites quantity was increased by 203.57 %,and the inhibition rate on A549 and SW-1990 were increased by 48.32 % and 48.32 % respectively.Then 150 L fermentation broth was obtained under the optimized conditions of fermentation.The fermentation broth was filtered by high speed centrifugation,obtaining mycelium and filtrate respectively.The filtrate was extracted with equal volume ethyl acetate 3 times,and got 80 g extract;Mycelium was extracted with ethanol in ultrasonic apparatus(30 min/time)for 3 times,about 20 g extract was got.We obtained about 100 g crude extract in the end.By various chromatographic methods,such as Sephadex LH-20 chromatography,reversion phase chromatography and HPLC chromatography,we separated the crude extract of the Penicillium sp.S-3-88 to get pure compound.Then compounds’ structure was identified on the basis of 1H and 13 C spectral data,MS data,and with the relevant literature.Finally,fifteen compunds were identified as: peniacetal(1),2,6-bis(1-phenyleth--yl)-phenol(2),2,4,6-tris(1-phenylethyl)phenol(3),2-acetyl-4(3H)quinazolinone(4),chrysogine(5),ergosterol peroxide(6),melithasterol B(7),22 E,24R-5α,6α-epoxyergosta-8(14),22-diene-3β,7α-diol(8),cerevisterol(9),benzo[d]-thiazol-2(3H)-one(10),5-methoxy-3,4-dihydro-naphthalen-1(2H)-one(11),5-hydroxy-3,4-dihydronaphthalen-1(2H)-one(12),octadecyl3-(3,5-di-tert-butyl-4-hydroxy)-phenylpropanoate(13),diisobutyl phthalate(14)and butyl-isobutyl-phthalate(BIP)(15).In these compounds,compound 1 is a new skeleton compound;compounds 2 and 3 are obtained from the polar Penicillium genus strains for the first time.Biological activities including antibacterial/fungus,cytotoxity and PTP1 B inhibitory activities of these compounds were evaluated.The results showed compound 2 inhibited PTP1 B activity obviously with IC50 of 14.81 ± 0.428 μM;but the IC50 of compound 3 was greater than 49.26 μM(IC50 of positive control oleanolic acid was 5.41 ± 0.61 μM).On the other hand,we firstly found that compounds 2 and 3 showed good inhibitory effect on HCT-116 cell proliferation,with IC50 of 5.09 ± 1.15 μM and 16.43 ± 2.31 μM respectively.The IC50 of compound 2 was lower than the positive control drug cisplatin and the effect of compound 3 was about the same with cisplatin(Cisplatin,IC50: 16.30 ± 0.70 μM).Compound 3 played good inhibibitory effect on SW-1990 cell proliferation(IC50: 9.47 ± 0.86 μM).Antibacterial/fungus activity evaluation showed that,compound 6 had certain inhibition against Staphylococcus aureus and Bacillus subtilis,with MIC of 50 μg·m L-1;Compound 4,6,7,8,10 and 13 showed strong inhibition effect on Pyricularia oryzae(MIC: 1.9 μg·mL-1).The structures of identified 15 compounds are as follows:... |
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