CN105296388A - Streptomyces koyangensis strain and anti-cancer active metabolite and application thereof - Google Patents

Streptomyces koyangensis strain and anti-cancer active metabolite and application thereof Download PDF

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CN105296388A
CN105296388A CN201510716704.3A CN201510716704A CN105296388A CN 105296388 A CN105296388 A CN 105296388A CN 201510716704 A CN201510716704 A CN 201510716704A CN 105296388 A CN105296388 A CN 105296388A
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streptomycete
cancer
bacterial strain
actinomycetes
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CN105296388B (en
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刘树林
周玉洁
沐晓芹
刘慧迪
赵丹丹
陈浩庭
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Harbin Engineering University
Harbin Medical University
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Abstract

The invention discloses a streptomyces koyangensis strain and an anti-cancer active metabolite and application thereof, and belongs to the technical field of biological pharmacy. The streptomyces koyangensis strain is named AD16 and named Streptomyces koyangensis in a classified mode, and the microbial preservation number is CGMCC No.11485. The streptomyces koyangensis strain AD16 is from the human intestinal tract, is actinomycetes separated newly, has characteristics different from those of other kinds of actinomycetes and is capable of generating the anti-cancer active metabolite, and the metabolite of the streptomyces koyangensis strain can be used for treating cancers. As the streptomyces koyangensis strain AD16 is found in the human intestinal tract and has little or no side effect on the human body, the problems that existing anti-cancer drugs have large side effects, and patient compliance is poor are solved, and it is expected that cancers can be prevented by adjusting intestinal flora balance.

Description

One plant height sun streptomycete bacterial strain and antitumour activity meta-bolites and application
Technical field
The present invention relates to a strain actinomycetes strain, be specifically related to the Gaoyang streptomycete bacterial strain that can produce antitumour activity meta-bolites that a strain is separated from human intestinal, and preparing the application in cancer therapy drug.The invention belongs to biological pharmacy technical field.
Background technology
Cancer is a kind of M & M disease all very high and in rising trend year by year, and cancer cells is the fundamental cause producing cancer, is the autogenous cell of variation, has the feature of indeterminate growth, transfer, is therefore difficult to control.Seriously endanger health and the life of the mankind, receive the concern of countries in the world.Estimate according to the World Health Organization, whole world cancer is ill about 7,000,000 people every year, dead 5,000,000 people.It is reported, between past 10 years, the M & M of global cancer increase about 22% [Bai Chunxue, lung cancer therapy new concept [J]. popular medicine, 2005,5 (5): 28; Soup seapeak, Yi Yanghua. ocean anti-neoplastic research overview and prospect [J]. medicine service and research, 2002,32 (1): 7-16; Wang Chuanxin. the progress [J] of tumor markers. Shandong health, 2006,12:53-54.], and continuation is also risen by this numeral, and the number expecting the death because of cancer stricken every year of the year two thousand thirty whole world will more than 13,100,000.The latest survey of 2012 yearly correlation departments shows, China is every year because the number of cancer mortality reaches 2,000,000.
In cancer therapy, although be primary selection with radical surgery, pharmacological agent is still a very important link, uses Effective Anti cancer drug can extend survival time and the quality of life of patient.Cancer therapy drug the most common has antimetabolite, natural plant, antitumor antibiotics etc. at present.In numerous antitumor drug, the proportion shared by natural plant kind antitumour drug is maximum, occupies the share of 27.0%, is secondly anti-metabolism antitumour drug, accounts for 26.1%.In the antitumour drug of first 10 of single product rank, plant antitumour drug occupies the first two seat, is respectively taxol and docetaxel.But, derive from the ebormycine of sorangium cellulosum (Sorangiumcelulosum) and the anticancer machine-processed basic simlarity of taxol, but it is more soluble in water than taxol, molecular weight is less, structure is simple, produce in synthesis and curative effect is all better than taxol, especially to the tumour cell of taxanes resistance, there is high reactivity, be considered to the renewal product [Li Zhifeng of taxol, EtienneNguimbi, Li Yuezhong, Deng. the PKS/NRPS heterozygous genes bunch [J] of ebormycine (Epothilones). biotechnology journal, 2003, 19 (5): 511-515.].This shows that microorganism can produce efficient anti-cancer active matter.Microorganism is the resource of important natural product, and it is numerous in variety, and the chemical structure of its meta-bolites is complicated and various, is the inexhaustible treasure-house of development new drug.Actinomycin be the mankind find the first there is the microbiotic [Hollstein of antitumous effect, U.Actinomycin.Chemistryandmechanismofaction.ChemicalRevi ews.1974, 74 (6): 625 – 652], it is the gram positive organism of a class high (G+C) %, it is jointly found [WaksmanSA by Sai Erman A Waksman and his colleague H.B. John Woodruff in 1940 the earliest, WoodruffHB.Actinomycesantibioticus, aNewSoilOrganismAntagonistictoPathogenicandNonpathogenic Bacteria [J] .JBacteriol, 1941, 42 (2): 231-249.].The active compound that actinomycetes produce comprises microbiotic, antiviral substance, immunosuppressor, antitumor drug and multiple enzyme.Derive from microbic activity compound so far, actinomycetic meta-bolites proportion is maximum, account for 45%, also have the meta-bolites biological activity of 10,000-3 ten thousand kinds of microorganisms not bright according to estimates, wherein actinomycetes source is 5000-10000 kind, therefore has very important significance for actinomycetic research.Research find actinomycin mainly through Cell differentiation inducing activity and apoptosis, suppress some protease activity and affect cell cycle etc. and play its anti-tumor activity.Actinomycin can suppress the mitotic division of the mammalian cell of vitro culture.Dactinomycin can cause the morphological change of HeLa cell and some mouse tumor cells, causes the fragmentation of kernel and chromosomal fracture.Dactinomycin can inducing mouse red corpuscle under low dosage, differentiation [the BoyneJR of mouse bone marrow cells red blood corpuscle stem cell, WhitehouseA.NucleolardisruptionimpairsKaposi ' ssarcoma-associatedherpesvirusORF57-mediatednuclearexpor tofintronlessviralmRNAs [J] .FEBSLett, 2009, 583 (22): 3549-3556.], it is combined with DNA simultaneously, RNA is suppressed to synthesize [J.M.Kirk., ThemdoeofactionofactinomycinD, Biochim.BioPhys.Acat (1960) 42:167 1, I.H.Golbderg, M.Rabinowitz, ActiomnycinDihnbitionofdeoxyribonucleicofacid-dependents ynthesisofribonueleicacid.Scienee (1962) 136:315 1, 1.H.Goldberg, M.Rabinowitz, E.Reieh, Basisofactinomycinaction, I.DNAbindingandinhibitionofRNA mono-polymerasesyntheticreactionsbyactinomycinProc.Natl.Acda. Sci-(1962) 48:2094-2101, E.Reieh, 1.H.Goldberg, ActinomycinandnucleicacidfuctionPorg.NucleicAcidRes.Mol. Biol. (1964) 3:183-234.] block RNA polymerase catalysis under the RNA that generates, block the extension stage [A.Sentenac of transcription, E.J.Simon, P.Formageot, InitiationofchainsbyRNApolymeraseandtheeffectsofihnibito rsstudiedbyadirectfiltrationttechnique.Biochim.BioPhys.A cta (1968) 161:299-308.].Dactinomycin can with double-stranded DNA mortise, suppress archaeal dna polymerase, but the restraining effect of archaeal dna polymerase be can not show a candle to suppress the effect of RNA polymerase.Dactinomycin has a strong impact on the synthesis of ribosome-RNA(rRNA), active cell apoptosis cascade reaction, cause apoptosis [R.P.Perry, selectiveeffectofactinomycinDontheintracellularofRNAsynt hesisintissueculturecells.ExP.CellRes. (1963) 29:400-406; S.IaPalucci mono-EsPinoza, M.T.Farnez-FenrandezEeffetofProteinsynthesisihnibitosran dlowconcentrationsofactinomycinDonribosomalRNAsynthesis, FEBSLett. (1979) 107 (2): 281-284.].Research also finds that nucleic acid metabolism is responsive to dactinomycin, find that it affects the interaction [R.H.Singer between mRNA and rrna, S.Penman, SatbiliytofHeLacellmRNAinactinomycin.Natuer, (1972) 240:100 1], have impact on DNA methylation [M.Gold, J.Hurwitz, TheenzymaticmethylationofribonucleicacidnadDeoxyribonucl eicacid.VI.FURTHERSTUDIESONTHEPROPERTEISOFTHEDEOXYRIBONU CLEICACIDMETHYLATIONREACTION.J.Biol.Chem. (1964) 239, 3866-3874.], reparation [the M.M.Elkind of DNA, GF:Whitmore, T.Alescio, ActinomycinD:SuppressionofRecoveryinX-irradiatedmammalia ncells.Science (1964) 143, 1454-1457.], degraded [the N.K.SarkareffectsofactinomycinDandmitomycinConthedegrati onofdeoxyribonucleicacidandpolydeoxyribonucleotidebydeox yribonucleasesandvenomphosphodiestearse.Bioehim.BioPhys. Acta (1967) 145 of deoxyribonuclease, 174-177], pyrophosphate exchange reaction [the H.Goldberg of ribose triphosphoric acid, M.Rabinowitz, E.Reich, Basisofactinomycinaction.II.Effectofaetinomycinonthenucl eosidetriphosphate-inorganicpyorphosphateexchange.Proc.N atl.Acad.Sci. (1963) 49, 226-229.] and RNA by nucleus to cytoplasmic transfer process [H.Harris, Rapidlylabelledribonucleicacidinthecellnucleus.Nature (1963) 198, 184-185, R.Lieberman, Abrams, P.Ove, ChangesinthemeatabolismofribonucleicacidPrecedingthesynt hesisofdeoxyribonucleicacidinmammaliancellsculturedfromt heanimal.J.Biol.Chem. (1963) 238:2141-2149, M.Girard, S.Penman, J.E.DarneII, TheeffectofactinomycinonribosomeFormationinHelacells.pro c.Natl.Acad.Sci. (1964) 51:205-211.].The dactinomycin of high density can cause the degraded [GAcs of RNA in eucaryon and prokaryotic cell prokaryocyte, E.Reieh, S.Valanju, RNAmetabolismofB.Subtilis, effectsofactinomycin.Biochim.BioPhys.Acta (1963) 76:68; H.Harris, Rapidlylabelledribonucleicacidinthecellnucleus.Natuer (1963) 198,184-185; R.Wiesner.GAes, E.Reieh, A.Shafiq, Degradationofribonucleicacidinmousefibroblaststreatedwit hactinomycin.J.CellBiol. (1965) 27,47-52; GA.Stewart, E.Fabre, Therapidaccelerationofhepaticnuclearribonucleicacidbreak downbyactinomycinbutnotbyethionine.J.Biol.Chem. (1968) 243,4479-4485.].The actinomycin AV of high density makes tumour cell cycle S and G2/M phase block [FieldsGB, NobleRL.Solidphasepeptidesynthesisutilizing9-fluorenylme thoxycarbonylaminoacids [J] .IntJPeptProteinRes, 1990,35:161-214.], illustrate that actinomycin AV may be at the thorough killing off tumor cells of the synthesis phase of DNA, can prevent simultaneously metastasis of cancer and diffusion [Shi Shanshan. the antitumor and antiviral study [D] of marine actinomycete element X2. Guangzhou: and south is greatly, 2009.].
Actinomycetes are extensive in distributed in nature, are mainly present in soil, empty G&W with spore or mycelia state, especially enrich with organism, in maximum in neutral or subalkaline soil.Because the research in recent years for oceanic resources is goed deep into, find the actinomycetes in ocean, enriched actinomycetic meta-bolites, too increase the chance finding anti-cancer active matter.Find that new actinomycetic active metabolite becomes the effective way finding new cancer therapy drug.In experimentation, inventor obtains strain actinomycetes from the fecal sample of children, and this is the actinomycetes that a strain obtains in human intestinal, and finds that it has significant antitumour activity.Therefore in the present invention, contriver utilizes actinomycetic active metabolite Therapeutic cancer.
The present invention utilizes the actinomycetic meta-bolites Therapeutic cancer in Healthy People enteron aisle, by carrying out the sample collected centrifugally obtaining supernatant liquor, and separation and Culture obtains actinomycetes, and to actinomycetes enlarged culturing, obtain its active metabolite, join in the cancer cells of Secondary Culture, cultivate 6-48 hour, in Microscopic observation cancer cells form, find that a large amount of cavity and fusion appear in cancer cells, and occur floating successively, dead.Illustrate that deriving from healthy HE actinomycetic meta-bolites has clear and definite antitumour activity.
Summary of the invention
Cancer has become the disease of serious threat human life health, because cancer cells belongs to autogenous cell, can infinite multiplication, conversion and transfer, therefore the treatment of cancer is very difficult.If can effectively contain developing of cancer, will be a significant benefit to the prolongation of average human life, and be conducive to the life and health of the mankind, the research therefore for Therapeutic cancer medicine is focus always.The cancer therapy drug applied comprises the synthetic chemistry analogue of inhibition tumor cell metabolism, the anti-cancer active matter, antitumor antibiotics etc. in plant extract source, and antitumor antibiotics wherein has vast potential for future development, actinomycetes are as a kind of microorganism that can produce anti-cancer active matter, important role in anticancer therapy, and new meta-bolites is had for anticancer for the actinomycetes in new source, derive from the actinomycetes in enteron aisle different from the source of classic Actinomycete, its meta-bolites can become new cancer therapy drug.
For above problem, the invention provides the Gaoyang streptomycete AD16 bacterial strain that a strain can produce antitumour activity meta-bolites, and the meta-bolites of this bacterial strain or this bacterial strain is preparing the application in cancer therapy drug.
Inventor screens the actinomycetes AD16 bacterial strain that a strain can produce antitumour activity meta-bolites from healthy children ight soil, Gaoyang streptomycete (Streptomyceskoyangensis) is initially identified as through morphologic observation and molecular biology research, called after AD16, Classification And Nomenclature is Streptomyceskoyangensis, microbial preservation number is CGMCCNo.11485, preservation date is on October 13rd, 2015, depositary institution is China Committee for Culture Collection of Microorganisms's common micro-organisms center, preservation address is No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, Institute of Microorganism, Academia Sinica.
The concrete steps that the present invention screens the streptomycete AD16 bacterial strain that can produce antitumour activity meta-bolites are:
(1) with Healthy People ight soil for sample, therefrom separation and Culture actinomycetes detect its antitumour activity.
(2) for being separated the actinomycetes enlarged culturing obtained, single bacterium colony is got in 5mL Gause I liquid nutrient medium, 160 revs/min, 16-40 DEG C from flat board, increase after 4-16 days, again 5mL bacterium liquid is transferred in 100mL Gause I liquid nutrient medium, 160 revs/min, 16-40 DEG C, a large amount of amplification 4-16 days, finally by centrifugal for bacterium liquid, the centrifugal 10min of 8000rpm/min, gets supernatant liquor and does the experiment of follow-up inhibiting tumor cell.Cancerous cell line used is HeLa cell.
(3) metamorphosis of cancer cells is observed, and record.
Streptomycete of the present invention (Streptomyceskoyangensis) AD16 bacterial strain has following characteristic:
(1) strain morphology feature: main in mycelial growth with the prokaryotic organism of sporogenesis, have substrate mycelium and aerial mycelium, spore presents white.
(2) 16SrRNA of bacterial strain molecular biological characteristics: strains A D16 measures 1338 bases altogether, compare by the 16SrRNA sequence of the Gaoyang streptomycete of having delivered in BLAST and GenBank, sequence identity reaches more than 98%, strains A D16 is Gaoyang streptomycete (Streptomyceskoyangensis), and the sequence of 16SrRNA is shown in SEQIDNo.1.
The meta-bolites preparation of Gaoyang of the present invention streptomycete (Streptomyceskoyangensis) AD16 bacterial strain: get single bacterium colony in 5mL Gause I liquid nutrient medium from flat board, 160 revs/min, 16-40 DEG C, increased after 4-16 days, then was transferred in 100mL Gause I liquid nutrient medium by 5mL bacterium liquid, 160 revs/min, 16-40 DEG C, a large amount of amplification 4-16 days, finally by centrifugal for bacterium liquid, the centrifugal 10min of 8000rpm/min, gets supernatant liquor and does the experiment of follow-up inhibiting tumor cell.
Compared with prior art, beneficial effect of the present invention is embodied in:
(1) streptomycete AD16 bacterium source of the present invention is in healthy human body enteron aisle, is a kind of actinomycetes of new source, possesses the character that the actinomycetes of other soil sources are different, can produce antitumour activity meta-bolites.
(2) because it finds in healthy human body enteron aisle, the side effect for human body can very low or nothing.
(3) pass through the difference regulating the state of human body enteric microorganism or the ratio of quantity, thus reach the object of preventing cancer.
Accompanying drawing explanation
Fig. 1 be streptomycete AD16 bacterial strain three road streak culture after bacterium colony result figure;
Fig. 2 is the morphological change of HeLa cell after the meta-bolites effect of streptomycete AD16 bacterial strain;
Fig. 3 is the scratch experiment result figure of HeLa cell after meta-bolites effect 24h, 48h of streptomycete AD16 bacterial strain;
Fig. 4 is the result figure of HeLa cell MTT after meta-bolites effect 24h, 48h, 72h of the streptomycete AD16 bacterial strain of different concns.
Embodiment
Below by experiment, also the present invention will be further described in conjunction with the embodiments, it should be understood that these embodiments only for the object of illustration, never limit the scope of the invention.
The screening of embodiment 1 streptomycete AD16 bacterial strain
1 materials and methods
1.1 substratum and solution
1) Gause I solid culture based formulas (1L):
Zulkovsky starch 20g, KNO 31g, K 2hPO 40.5g, MgSO 47H 2o0.5g, NaCl0.5g, FeSO 47H 2o0.01g (being made into mother liquor to add again), agar 20g;
First dissolve Zulkovsky starch, add gauge water and fully dissolve, then add other compositions, remember after high pressure to shake up.
2) potassium bichromate: concentration 50mg/L (during screening use, other need not)
Stock solution: 5g/L (matching while using, lucifuge, filtration sterilization) (not contacting, poisonous);
If configuration 100mL Gause I solid medium, add 1mL potassium bichromate storage liquid (adding when substratum is not too hot, mixing).
3) FeSO 47H 2o mother liquor: 0.01g/mL
1gFeSO 47H 2o is dissolved in 100mLH 2in O, so 1000mL Gause I solid medium adds 1mL ferrous sulfate mother liquor.
4) PBS buffer formulation (1L):
KH 2PO 40.27g，Na 2HPO 41.42g，NaCl8g，KCl0.2g。
The acquisition of 1.2 streptomycete AD16 bacterial strains
(1) with Healthy People ight soil for sample, therefrom separation and Culture actinomycetes.
Sampling explanation
We will give every ight soil supplier one sampling bag, comprise following article:
50mL centrifuge tube x1
Spoon x1
PE gloves x2
Paper disc x1
Informed Consent Form x1
X1 is said in sampling explanation
Operation instructions:
(1) read Informed Consent Form in detail, ight soil supplier signature also fills in essential information in detail.
(2) please ight soil is discharged in disposable paper disc as far as possible.
(3) bring PE gloves, take ight soil centre portions with spoon, about 10g (approximately reaching the scale marks place of centrifuge tube 10mL), puts in 50mL centrifuge tube, after tightening centrifuge tube lid, puts back in valve bag.
(4) whole operating process is avoided touching other uncorrelated article as far as possible.
(5) because morning Monday submits sample, so collect the ight soil in morning Monday as far as possible, fresh to ensure.
(6) please centrifuge tube and Informed Consent Form that ight soil is housed are loaded valve bag, returned by lab assistant centralized collection.
(2) actinomycetic separation and Culture:
1. sample, numbering: 1,2,3,4 Add equal-volume sterile distilled water, centrifugal: 8000rpm, 30min.Collect supernatant liquor and sample pellet;
2. sample pellet is mixed with the sand after autoclaving; Shady and cool place natural air drying 4-16 days.
3. pulverize, 80 DEG C of xeothermic 1h of constant temperature;
4. get the sample that 1g processes and be dissolved in 10mLPBS damping fluid the 30min that vibrates;
5. leave standstill 30s;
6. get supernatant and carry out 10 times, 100 times, 1000 times dilutions;
7. get 10 -2, 10 -3pipe 100 μ l is coated with flat board respectively (containing potassium bichromate +gause I solid medium), cultivate for 0-40 DEG C; Get single bacterium colony be placed in slide glass mixes with suitable quantity of water after covered, 40 × visible mycelium spore, bacterium colony is hard, not easily picking.
8. single bacterium colony three road is streak culture, 28 DEG C of cultivations, and the bacterium colony result figure after streptomycete AD16 bacterial strain three road is streak culture is shown in Fig. 1;
9. get the close painting of mono-clonal, 28 DEG C of cultivations from the flat board of three road line, select the close painting of the representational bacterium colony of form and cultivate;
10. deposit bacterium: 500 μ l50% glycerine+500 μ l Gause I liquid nutrient mediums, after mixing, deposit bacterium;
11. inclined-planes: inoculating needle picking colony inserts inclined-plane (in 1.5mLEP pipe, having Gause I culture medium slant), repeatedly inserts, more repeatedly streak culture for subsequent use on inclined-plane.
(3) for being separated the actinomycetes enlarged culturing obtained:
Actinomycetes are inoculated in Gause I liquid nutrient medium enlarged culturing, 160 revs/min, 16-40 DEG C, incubation time: 4-16 days.
(4) cultivate HeLa cell, Secondary Culture and bed board, every hole 100 μ l, cell density is 1x10 5/ mL, after HeLa cell attachment, add the meta-bolites 5 μ l (supernatant liquor that streptomycete AD16 cultivated through 4-16 days) of streptomycete AD16 bacterial strain wherein, control group is isopyknic Gause I liquid nutrient medium, then cultivates observation after 6-48 hour.Microscopic observation HeLa cellular form.There is a large amount of cavity and fusion in HeLa cell, sees Fig. 2.
The meta-bolites preparation of streptomycete AD16 bacterial strain: get single bacterium colony in 5mL Gause I liquid nutrient medium from flat board, 160 revs/min, 16-40 DEG C, increased after 4-16 days, then was transferred in 100mL Gause I liquid nutrient medium by 5mL bacterium liquid, 160 revs/min, 16-40 DEG C, a large amount of amplification 4-16 days, finally by centrifugal for bacterium liquid, the centrifugal 10min of 8000rpm/min, gets supernatant liquor and does the experiment of follow-up inhibiting tumor cell.
(5) cultivate HeLa cell, carry out scratch experiment, in 100 μ l cell culture fluids, add the meta-bolites 100 μ l of AD16 bacterial strain.As seen from Figure 3, the cancer cells of control group is to cut place Invasion and Metastasis, and the experimental group of meta-bolites effect 24h through streptomycete AD16 bacterial strain, find that the Invasion and Metastasis ability of cancer cells to cut is much smaller than control group, through the meta-bolites effect 48h of streptomycete AD16 bacterial strain, HeLa cell presents dead floating state, and control group cut now heals substantially.The Invasion and Metastasis ability of HeLa cell effectively can be suppressed by the meta-bolites demonstrating streptomycete AD16 bacterial strain that scratch experiment is strong.
(6) cultivate HeLa cell, Secondary Culture and bed board, every hole 100 μ l, cell density is 1x10 5/ mL, meta-bolites 5 μ l, 10 μ l, 20 μ l, 40 μ l, 80 μ l, the 160 μ l of streptomycete AD16 bacterial strain are added wherein respectively after HeLa cell attachment, control group is Gause I liquid nutrient medium, respectively establishes 6 repeating holes, then cultivates observation after 6-72 hour.Every hole adds 20 μ lMTT solution, continues to cultivate 4h.Stop cultivating, carefully suck nutrient solution in hole.Every hole adds 150 μ l dimethyl sulfoxide (DMSO), puts low-speed oscillation 10min on shaking table, and Jie Jing Wu Za is fully dissolved.The light absorption value in each hole is measured at enzyme-linked immunosorbent assay instrument OD492nm place.As shown in Figure 4, the meta-bolites of streptomycete AD16 all presents gradient for the restraining effect of HeLa cell and time and concentration to result.
Finally, inventor filters out a strain has inhibit activities actinomycetes AD16 bacterial strain to tumour cell from Healthy People enteron aisle.
The classification position of embodiment 2 streptomycete AD16 bacterial strain
(1) morphological specificity is observed: main in mycelial growth with the prokaryotic organism of sporogenesis, have substrate mycelium and aerial mycelium, spore presents white.
(2) molecular biology identification: streptomycete AD16 bacterial strain is carried out three roads streak culture after, mailing carries out 16srRNA order-checking and qualification to Jin Wei intelligence bio tech ltd, the 16SrRNA of strains A D16 measures 1338 bases altogether, compare by the 16SrRNA sequence of the Gaoyang streptomycete of having delivered in BLAST and GenBank, sequence identity reaches more than 98%.Therefore, strains A D16 is accredited as and Gaoyang streptomycete (the streptomyces bacterium of the nearly edge of Streptomyceskoyangensis, called after AD16, Classification And Nomenclature is Streptomyceskoyangensis, microbial preservation number is CGMCCNo.11485, and preservation date is on October 13rd, 2015, and depositary institution is China Committee for Culture Collection of Microorganisms's common micro-organisms center, preservation address is No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, Institute of Microorganism, Academia Sinica.The sequence of 16SrRNA is shown in SEQIDNo.1.
The foregoing is only the preferred embodiments of the present invention, is only illustrative for the purpose of the present invention, and nonrestrictive; Those of ordinary skill in the art understand, and can carry out many changes in the spirit and scope that the claims in the present invention limit to it, amendment, and even equivalence is changed, but all will fall within the scope of protection of the present invention.

Claims (3)

1. plant height sun streptomycete (Streptomyceskoyangensis) bacterial strain, called after AD16, microbial preservation number is CGMCCNo.11485, and preservation date is on October 13rd, 2015, and depositary institution is China Committee for Culture Collection of Microorganisms's common micro-organisms center.
2. Gaoyang according to claim 1 streptomycete AD16 bacterial strain is preparing the application in cancer therapy drug.
3. the meta-bolites of Gaoyang according to claim 1 streptomycete AD16 bacterial strain is preparing the application in cancer therapy drug.
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CN105969685A (en) * 2016-05-12 2016-09-28 南昌大学 Method for screening microorganisms with anticancer activity in excrement
CN109306331A (en) * 2018-08-01 2019-02-05 西北民族大学 The bull streptomycete bacterial strain screening method and identification method of one plant of production anti-cancer active matter
CN110283762A (en) * 2019-08-01 2019-09-27 哈尔滨天齐人类第二基因组技术开发应用科技有限责任公司 One plant has the active streptomycete CCPM7649 of powerful anticancer and its application
CN110283762B (en) * 2019-08-01 2020-09-22 哈尔滨天齐人类第二基因组技术开发应用科技有限责任公司 Streptomycete CCPM7649 with strong anticancer activity and application thereof

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