CN102363755A - Streptomyces and method thereof for preparaing adriamycin - Google Patents

Streptomyces and method thereof for preparaing adriamycin Download PDF

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CN102363755A
CN102363755A CN2011101431773A CN201110143177A CN102363755A CN 102363755 A CN102363755 A CN 102363755A CN 2011101431773 A CN2011101431773 A CN 2011101431773A CN 201110143177 A CN201110143177 A CN 201110143177A CN 102363755 A CN102363755 A CN 102363755A
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zorubicin
streptomycete
preparation
cgmcc4827
streptomyces
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CN102363755B (en
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陈华
叶美丽
郑玲辉
王玲萍
白骅
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Zhejiang Hisun Pharmaceutical Co Ltd
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Zhejiang Hisun Pharmaceutical Co Ltd
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Abstract

The invention discloses a novel Streptomyces and a method for preparaing adriamycin or analogues thereof through culturing the Streptomyces. The Streptomyces sp.H323(CGMCC4827) disclosed by the invention is a brand new strain which can produce the adriamycin or the analogues of the adriamycin through fermentation in nutrient media containing assimilable carbon resources and nitrogen sources under an oxygen introducing condition.

Description

A kind of streptomycete and the method for producing Zorubicin thereof
Technical field
The present invention relates to a kind of new streptomycete, produce the method for Zorubicin or its analogue through this bacterium of fermentation culture.
Background technology
Anthracycline antibiotics (like Zorubicin, daunorubicin, pidorubicin) is the antitumor drug of widespread use.The structure of Zorubicin and daunorubicin basic identical (difference is only on a hydroxyl, as shown in the formula I), daunorubicin is only effective to acute leukemia (leukemia), and Zorubicin is then very wide to the therapeutic domain of malignant tumour.
Figure BSA00000507376100011
? ?R 1 R 2 R 3
Daunorubicin ?C(O)CH 3 H OH
Zorubicin ?C(O)CH 2OH H OH
Pidorubicin ?C(O)CH 3OH OH H
1974, U.S. Pat 3,803,124 disclose Zorubicin can be made through chemistry is semi-synthetic by the tunning daunorubicin.At present, semi-synthetic through daunorubicin is the method for suitability for industrialized production Zorubicin.Gondola Pharmacia & Upjohn S.P.A discloses a kind of method that is prepared Zorubicin by daunorubicin through enzymatic conversion method in Chinese patent CN1147835A.In addition, U.S. Pat 3,590,028 discloses a strain Zorubicin produces bacterial classification Streptomyces peucetius subsp.caesius ATCCNo.27952.U.S. Pat 4,012,284 disclose an other strain Zorubicin produces bacterial classification Streptomyces peucetius ATCCNo.29050.
Though there is the report Zorubicin to produce bacterial classification in the prior art, also rest on laboratory stage, do not form industrialization.Therefore, searching can produce Zorubicin new microbe bacterial classification and new preparing method's work still in proceeding.
Summary of the invention
One of the object of the invention is to provide a kind of new microbial strains that can produce Zorubicin or its analogue, a kind of new streptomycete H323, and its deposit number is CGMCC4827.
The present invention also provides new streptomycete H323 (CGMCC4827) (Streptomyces sp.H323), and it can be applied to produce Zorubicin or its analogue, perhaps is applied to produce the pharmaceutical composition that contains Zorubicin.
The present invention also provides a kind of employing streptomycete H323 (CGMCC4827) to prepare the method for Zorubicin or its analogue.This method comprises that employing streptomycete H323 (CGMCC4827) in the nutritional medium that contains assimilable carbon, nitrogenous source, carries out the process of aerobic fermentation.
Above-mentioned assimilable carbon source is selected from the combination of one of starch, dextrin, glucose, industrial molasses, glycerine, sucrose, lactose, SANMALT-S, trehalose, xylan, N.F,USP MANNITOL, sorbyl alcohol or above-mentioned substance.
Above-mentioned assimilable nitrogenous source is selected from the combination of one of yeast extract, yeast powder, beef extract, Tryptones, peptone, soybean cake powder, cottonseed meal, groundnut meal, seitan powder, Dried Corn Steep Liquor Powder, wheat bran, urea, ammonium salt or above-mentioned substance.
The temperature of said fermentation culture is 20 ℃-40 ℃, and preferred 25 ℃-30 ℃, pH is between the 6.5-7.5, preferred about 6.8; Incubation time is 144-216 hour; Oxygen-supply quantity is 0.5-1.0vvm.
Said fermentation mode is a liquid submerged fermentation.
Zorubicin can detect through following method:
Chromatographic column is the C18 post, 5 μ m, 4.6 * 250mm;
Moving phase: damping fluid: acetonitrile: methyl alcohol=500: 500: 60;
Damping fluid: get sodium lauryl sulphate 1.44g and phosphoric acid 0.68ml and be dissolved in the 500ml ultrapure water;
Flow velocity: 1.35ml/min;
Detect wavelength: 254mn;
Sample size: 10 μ l.
The Zorubicin that adopted among the present invention produces the mutant strain that bacterial classification can also obtain for the spontaneous mutant strain of streptomycete H323 (CGMCC4827), streptomycete H323 (CGMCC4827) or the mutagenesis through routine, and it is streptomycete H323 (CGMCC4827) that preferred Zorubicin produces bacterium.
Streptomycete H323 (Streptomyces sp.) microbial strains (CGMCC4827) is stored in China Committee for Culture Collection of Microorganisms common micro-organisms center on May 4th, 2011
(address: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, Institute of Microorganism, Academia Sinica), preserving number is CGMCC4827, and registers on the books, and proves survival.
The main biological property of above-mentioned bacterial strains is: the orange colour cast of colony colour is red, and colonial morphology is than rounding, and there is the fold projection on the surface, and diameter is medium-sized (about about 6mm), no spore, and substrate mycelium is flourishing, and mycelia combines with substratum closely, is difficult for provoking.
The invention describes the morphology of Streptomyces sp.H323 and the characteristic on the molecular level; Process and known Zorubicin produce bacterium and carry out the comparison of morphology and molecular level; Can assert that Streptomyces sp.H323 belongs to streptomyces; But it is different from known Zorubicin and produces bacterium Streptomyces peucetius subsp.caesius ATCC No.27952, Streptomycespeucetius ATCC No.29050, but a kind of brand-new bacterial classification.
The present invention compares with former technology and has the following advantages:
1. simplification production technique.Bacterial classification according to the invention can be produced Zorubicin or its analogue through the direct fermentation method, has saved from daunorubicin and has obtained the technology of Zorubicin through chemosynthesis, helps reducing production costs and reducing environmental pollution.
2. raising fermentation unit.The output that bacterial classification according to the invention produces Zorubicin or its analogue has increased significantly than original strain, helps realizing suitability for industrialized production.
Description of drawings:
Fig. 1 is the colonial morphology of bacterial strain of the present invention;
The a:ISP2 substratum adds trace element.
The b:ISP3 substratum.
The c:ISP4 substratum.
The d:ISP2 substratum.
Fig. 2 is the mycelia form of bacterial strain of the present invention;
A: * 400 times of mycelia forms.
B: * 1000 times of mycelia forms.
Fig. 3 for the fermenting process curve of bacterial strain of the present invention in shaking bottle (■-: Zorubicin output; total reducing sugar;-△-: reducing sugar;--: the pH value);
Fig. 4 is the HPLC collection of illustrative plates of the Zorubicin of bacterial strain generation of the present invention;
Fig. 5 analyzes collection of illustrative plates for the ESI/MS of the Zorubicin that bacterial strain of the present invention produces;
The Zorubicin that Fig. 6 produces for bacterial strain of the present invention 1The H-NMR collection of illustrative plates;
The Zorubicin that Fig. 7 produces for bacterial strain of the present invention 13The C-NMR collection of illustrative plates.
Embodiment
Embodiment 1: bacterium source
Streptomyces sp.H323 system separates the original strain that obtains from Taizhou, Zhejiang Province city one discarded farm soil of coastal area of southeastern China, obtain Zorubicin through mutagenic and breeding and produce bacterial strain.This bacterial strain in the ISP2 slant medium 28 ℃ cultivate after 10-12 days, under aseptic condition, shovel mycelium scraped with inoculation, in rub oral examination tube, grind mycelium and be suspended in the sterilized water, make bacteria suspension, confession NTG (nitrosoguanidine) mutagenic treatment.
Take by weighing NTG crystal 10mg, be dissolved in the aseptic Tris damping fluid of 10ml (pH8.0), add the 1ml bacteria suspension with transfer pipet again, place oscillation treatment 30min on 28 ℃ of rotary or reciprocating type shaking tables of substratum.The bacteria suspension of handling is coated on the ISP2 flat board through suitably diluting.Do not make the bacteria suspension of mutagenic treatment and also coat the dull and stereotyped conduct contrast of ISP2 through suitably diluting.Cultivate after 10 days inspection colony count, and meter lethality rate for 28 ℃.
Select through single bacterium colony 1000 strains after the NTG mutagenesis, carry out shake flask fermentation, HPLC detects Zorubicin output.The result filters out enhanced variant Streptomyces sp.H323.
Embodiment 2: strain identification
The main biological property of this bacterial strain is: the orange colour cast of colony colour is red, and colonial morphology is than rounding, and there is the fold projection on the surface, and diameter is medium-sized (about about 6mm), no spore, and substrate mycelium is flourishing, and mycelia combines with substratum closely, is difficult for provoking.Concrete colonial morphology and mycelia form are as depicted in figs. 1 and 2.
Design following primer according to actinomycetes 16S rRNA conserved sequence, and give birth to worker bio-engineering corporation by Shanghai and synthesize:
Upstream primer: 5 '-AGAGTTTGATCCTGGCTCAG-3 '
Downstream primer: 5 '-AAGGA GGTGATCCAGCCGCA-3 '
Utilize above-mentioned primer that the 16S rDNA sequence of this bacterial strain is carried out pcr amplification, the PCR system is:
Figure BSA00000507376100041
The PCR cycling condition: 95 ℃ of preparatory sex change 5min, 95 ℃ of sex change 1min of loop parameter, 52 ℃ of renaturation 45s, 72 ℃ are extended 1.5min, repeat 35 circulations after, 72 ℃ are extended 10min again, last 4 ℃ of insulations.Detect the PCR product with 0.8% agarose gel electrophoresis.Select for use band clearly product carry out purifying.
Amplified production reclaims through gel electrophoresis, and checks order after being connected to the T carrier, has obtained the sequence of this bacterial strain 16S rDNA primary structure.Through in the Genebank DB, carrying out similarity searching (blast), the result finds that the 16S rDNA sequence homology of itself and ripple plug streptomycete mutation Streptomyces peucetius subsp.caesius ATCC No.27952 is 96%.With the 16S rDNA sequence homology of another strain ripple plug streptomycete Streptomyces peucetius ATCC No.29050 be 97%.
In sum, H323 belongs to streptomyces, but it is different from known Zorubicin generation bacterium Streptomyces peucetiussubsp.caesius ATCC No.27952 and Streptomyces peucetius ATCC No.29050.Streptomyces sp.H323 is the brand-new bacterial classification of a strain.
Embodiment 3: the preparation Zorubicin
(1) mycelial preparation in inclined-plane and cultivation
Slant pore culture medium prescription (g/L): yeast extract 4, Fructus Hordei Germinatus extract 10, glucose 4; Agar 20, the preceding pH6.5-6.8 that disappears, test tube 30 * 200mm; Loading amount 20mL after 20 minutes, is cooled to 50-60 ℃ of pendulum inclined-plane through 121 ℃ of sterilizations; After 10 days, mycelia is ripe through 28 ± 1 ℃ of cultivations for access one ring mycelium.
(2) preparation of seed liquor and cultivation
Seed culture based formulas (g/L): Zulkovsky starch 30, glucose 10, soybean cake powder 20, CaCO 32, NaCl 3, the preceding pH6.5 that disappears, and shaking bottled liquid measure was the bottled 25mL of 250mL triangle, through 121 ℃ of sterilizations 20 minutes.The thalline inoculum size is 10 5-10 6C.f.u./mL, 28 ± 1 ℃ of culture temperature, 250rpm, shaking table shaking culture 48 hours, nutrient solution pH7.0-7.5 at this moment, mycelial concentration is 15-20% (volume percent).
(3) preparation of Zorubicin fermention medium and cultivation
Fermentative medium formula (g/L): W-Gum 80, yeast powder 30, CaCO 33, NaCl 3, the preceding pH6.8 that disappears, and shaking bottled liquid measure was the bottled 25mL of 250mL triangle, through 121 ℃ of sterilizations 20 minutes.The inoculum size of seed liquor with 5-10% (volume percent) inserted, at 28 ± 1 ℃, 250rpm shaking table shaking culture 7 days, HPLC surveys Zorubicin unit after the fermentation ends, and the unit of recording is 200mg/L.
Embodiment 4: the preparation Zorubicin
(1) preparation of seeding tank seed liquor
In the 20L seeding tank, drop into the seed culture medium (proportioning of seed culture medium is seen embodiment 3, adds 0.3% soya-bean oil simultaneously and makes skimmer) of 15L, steam sterilizing is adopted in sterilization; Following 30 minutes of 121 ℃ of conditions after cooling, insert the 200ml shake-flask seed liquid; 28 ± 1 ℃ of culture temperature, mixing speed 200rpm, air flow 1vvm; Cultivated 48 hours, this moment pH7.0-7.5, mycelial concentration is 15-25%.
(2) preparation of fermentation tank culture medium and cultivation
The prescription of fermention medium is identical with previous embodiment 3, but will add 0.05%PPG as skimmer, and fermentor tank 50L, the volume that feeds intake are 35L; The preceding pH6.8 that disappears, the back pH7.0 that disappears is in 121 ℃ of following steam sterilizings 30 minutes, after the cooling; Insert about 3.5L seed tank culture liquid, 28 ± 1 ℃ of leavening temperatures, rotating speed of agitator 150-200rpm, air flow are 0.8-1.0vvm; Fermentation culture 7 days is put jar, and recording the Zorubicin fermentation unit is 300mg/L.
Embodiment 5: the preparation Zorubicin
Seed culture medium (g/L): glucose 20, Zulkovsky starch 20, yeast powder 10, yeast extract 5, corn starch 5, soybean cake powder 10, CaCO 34, MgSO 47H 2O 1, and NaCl 3, pH6.5, and shaking bottled liquid measure was the 30mL/250mL triangular flask, through 121 ℃ of sterilizations 30 minutes.Dig ferfas filament access seed culture medium from the inclined-plane of embodiment 3, place 28 ℃ of cultivation 45hr on the shaking table.The seed culture fluid of 2.5ml inserts fermention medium (g/L) then: maltodextrin 100, and soybean cake powder 30, seitan powder 2.5, NaCl 2, CaCO 33, FeSO 47H 2O 0.001, MnCl 24H 2O 0.003, CoCl 26H 2O 0.01, ZnSO 47H 2O0.005, pH6.8.The fermention medium loading amount is the 25mL/250mL triangular flask, and 121 ℃ of sterilizations 30 minutes place on the shaking table 28 ℃ to cultivate 7 days.HPLC detects, and fermentation unit is 280mg/L.
Embodiment 6: the preparation Zorubicin
The preparation of seed culture fluid is carried out according to embodiment 5.The seed culture fluid of 2.5ml inserts fermention medium (g/L) then: starch 100, and yeast powder 30, soybean cake powder 3, NaCl 2, CaCO 33, FeSO 47H 2O 0.001, MnCl 24H 2O 0.003, ZnSO 47H 2O 0.005, pH6.8.The fermention medium loading amount is the 25mL/250mL triangular flask, and 121 ℃ of sterilizations 30 minutes place on the shaking table 28 ℃ to cultivate 7 days.HPLC detects, and fermentation unit is 300mg/L.
Embodiment 7: preparation Zorubicin analogue
The preparation of seed culture fluid and fermention medium and cultivation are carried out according to embodiment 3.HPLC surveys Zorubicin analogue unit after the fermentation ends, and wherein daunorubicin is 60mg/L, and two hydrogen daunorubicins are 20mg/L.

Claims (9)

1. a streptomycete H323 has the ability that produces Zorubicin or its analogue.
2. streptomycete H323 according to claim 1 is characterized in that its deposit number is CGMCC4827.
3. streptomycete H323 according to claim 2 (CGMCC4827), the application in preparation Zorubicin or its analogue.
4. the method for a fermentative prodn Zorubicin or its analogue is characterized in that: comprise and adopt streptomycete H323 in the nutritional medium that contains assimilable carbon source and nitrogenous source, carry out the step of aerobic fermentation.
5. preparation method according to claim 4 is characterized in that: said assimilable carbon source is selected from the combination of one of starch, dextrin, glucose, industrial molasses, glycerine, sucrose, lactose, SANMALT-S, trehalose, xylan, N.F,USP MANNITOL, sorbyl alcohol or above-mentioned substance.
6. preparation method according to claim 4 is characterized in that: said assimilable nitrogenous source is selected from the combination of one of yeast extract, yeast powder, beef extract, Tryptones, peptone, soybean cake powder, cottonseed meal, groundnut meal, seitan powder, Dried Corn Steep Liquor Powder, wheat bran, urea, ammonium salt or above-mentioned substance.
7. according to any one described preparation method of claim 4～6, it is characterized in that: it is the mutant strain that spontaneous mutant strain of streptomycete H323 (CGMCC4827), streptomycete H323 (CGMCC4827) or the mutagenesis through routine obtain that the Zorubicin that wherein adopted produces bacterium.
8. preparation method according to claim 7 is characterized in that: it is streptomycete H323 (CGMCC4827) that the Zorubicin that is wherein adopted produces bacterium.
9. preparation method according to claim 4 is characterized in that: the temperature of said fermentation culture is 20 ℃-40 ℃, and pH is between the 6.5-7.5, and incubation time is 144-216 hour.
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Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103087124A (en) * 2012-11-21 2013-05-08 浙江海正药业股份有限公司 Method for preparing high-purity adriamycin
CN105624239A (en) * 2014-10-29 2016-06-01 浙江海正药业股份有限公司 Method for producing epirubicin
CN105861455A (en) * 2016-05-16 2016-08-17 浙江海正药业股份有限公司 DoxA protein mutant as well as encoding gene and application thereof
CN107541481A (en) * 2016-06-23 2018-01-05 上海医药工业研究院 A kind of genetic engineering bacterium for producing Epi-ADM and its application
CN113293187A (en) * 2021-06-23 2021-08-24 道中道（菏泽）制药有限公司 Fermentation liquor pretreatment method for doxorubicin production

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3590028A (en) * 1967-04-18 1971-06-29 Farmaceutici Italia Adriamycin derivatives
US4012284A (en) * 1962-11-16 1977-03-15 Societa' Farmaceutici Italia, S.p.A. Process of preparation of antibiotic F.I. 1762 derivatives
CN101144085A (en) * 2006-09-15 2008-03-19 上海医药工业研究院 Anthracene nucleus antibiotic producing strain dnrX gene fragment, gene breaking engineering bacterium prepared from the same and application thereof

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4012284A (en) * 1962-11-16 1977-03-15 Societa' Farmaceutici Italia, S.p.A. Process of preparation of antibiotic F.I. 1762 derivatives
US3590028A (en) * 1967-04-18 1971-06-29 Farmaceutici Italia Adriamycin derivatives
CN101144085A (en) * 2006-09-15 2008-03-19 上海医药工业研究院 Anthracene nucleus antibiotic producing strain dnrX gene fragment, gene breaking engineering bacterium prepared from the same and application thereof

Cited By (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103087124A (en) * 2012-11-21 2013-05-08 浙江海正药业股份有限公司 Method for preparing high-purity adriamycin
CN103087124B (en) * 2012-11-21 2016-01-13 浙江海正药业股份有限公司 A kind of method preparing Zorubicin
CN105624239A (en) * 2014-10-29 2016-06-01 浙江海正药业股份有限公司 Method for producing epirubicin
CN105861455A (en) * 2016-05-16 2016-08-17 浙江海正药业股份有限公司 DoxA protein mutant as well as encoding gene and application thereof
WO2017198085A1 (en) * 2016-05-16 2017-11-23 浙江海正药业股份有限公司 Doxa protein mutant, and coding gene and applications thereof
CN105861455B (en) * 2016-05-16 2018-10-26 浙江海正药业股份有限公司 DoxA protein mutants and its encoding gene and application
CN107541481A (en) * 2016-06-23 2018-01-05 上海医药工业研究院 A kind of genetic engineering bacterium for producing Epi-ADM and its application
CN107541481B (en) * 2016-06-23 2021-07-02 上海医药工业研究院 Genetic engineering bacterium for producing epirubicin and application thereof
CN113293187A (en) * 2021-06-23 2021-08-24 道中道（菏泽）制药有限公司 Fermentation liquor pretreatment method for doxorubicin production

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