LV10500B - Method for producing of complex antibiotic- cyclosporin and/or it's components and method for producing of tolypocladium varium culture - Google Patents

Method for producing of complex antibiotic- cyclosporin and/or it's components and method for producing of tolypocladium varium culture Download PDF

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Publication number
LV10500B
LV10500B LVP-93-708A LV930708A LV10500B LV 10500 B LV10500 B LV 10500B LV 930708 A LV930708 A LV 930708A LV 10500 B LV10500 B LV 10500B
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Latvia
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varium
producing
components
tolypocladium
cyclosporin
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LVP-93-708A
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LV10500A (lv
Inventor
Jekkel-Nee-Bokany Antonia
Ambrus Gabor
Toth-Sarudy Eva
Szabo Istvan Mihaly
Helber-Nee-Dobos Agota
Andor Attila
Albrecht Karoly
Kinczil Kalman
Szell Valeria
Tomori-Nee-Joszt Eva
Moravcsik Imre
Polya Kalman
Erdei Janos
Kiss Lajos
Makadi Bela
Nagy Karoly
Palotas Bela
Deli Nee Konszky Etelka
Buzasi Karoly
Molnar-Nee-Antal Aniko
Santha Gy Rgy
Szaszhegyesi Vilma
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Biogal Gyogyszergyar
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Publication of LV10500B publication Critical patent/LV10500B/lv

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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/14—Fungi; Culture media therefor
    • C12N1/145—Fungal isolates
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A61P37/02—Immunomodulators
    • A61P37/06—Immunosuppressants, e.g. drugs for graft rejection
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K7/00—Peptides having 5 to 20 amino acids in a fully defined sequence; Derivatives thereof
    • C07K7/64—Cyclic peptides containing only normal peptide links
    • C07K7/645—Cyclosporins; Related peptides
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
    • C12R2001/00—Microorganisms ; Processes using microorganisms
    • C12R2001/645—Fungi ; Processes using fungi
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
    • Y10S—TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y10S435/00—Chemistry: molecular biology and microbiology
    • Y10S435/8215—Microorganisms
    • Y10S435/911—Microorganisms using fungi

Description

LV 10500 CII0C0B nOJD^EHHH ΚΟΜΓΗΕΚΟΗΟΓΟ ΑΗΤΗΕΗΟΤΗΚΑ, ΠΙΓΑΜΜ
OnncaHHe iboāpeTeHHe othochtch k enoeo6y πολώθηκα κομπλθκοηογο
aHTMOHOTMKa U^ĪKJIOCnopHHa Η βΓΟ Κ0ΜΠ0Η6ΗΤ0Β UHKJIOCnopHHa A, UHKJīocnopHHa B h UHKJiocnopHHa C, aapočHOH $epMeHTaiiwefL
Oraejiī)Kfaie κομγγοηθητη KOMiuieKca npe^cTaBJiHiOT cocTOHinne H3 II aMMHOKMCJioT i^iKjnraeeKiie 3 HetfTpajībHbie, ηθποληρηηθ ojraronen-THflbi, KOToptie oTJiM^aroTCH spyr ot flpyra tojibko o^hoh vum sb^mh y^aCTByfOUiHMPI B ΠΟΟΤρΟβΗΗΗ aMKHOKKCJIOTaMM.
Jfo ΚΟΜΠΟΗβΗΤΟΒ ΚΟΜΠΛβΚΟΗΟΓΟ aHTMČHOTMKa IļHKJIOCĪIOpHH A H UHKJiocnopHH C ābīJiH BbiflejieHH cna^ajia A. Ruegger m ap. (Nelu. Chim. Acta 59: 1075 /1975/) u iļMKJiocnopHHH B, D, E m C R. Traber h ap. (Helv. Chim. Acta 60: 1568 /1977/; naTeHT Bejn>-rm N879 402 M3 Kyjn»TypH ΗΑθΗΤΗφκΐΰφΟΒΒΗΗΟΓΟ mm paHMie KaK Trichoderma polysporum (Bink ex Pērs.) Rifai rpnSKa (NRRL 8044). ΠοδβΗββ obiJia yToqHeHa cMCTeMaTbmecKan KJiaccH$HKaipia co-OTBeTCTByiomero niTaMMa rpMČKa (NRRL 8044), m c τθχ nop oh dbui KasBaH b JiHTepaType KaK Tolypocladium inflatum Gams. B HacTOHmee ΒρβΜΗ μββθοτηη yste 25 mncjiocnopHHOB, KOTopbie očosHcik'aiOTCH 6yKBaMM ot A ro Z (Helv. Chim. Acta 70, 13 - 2 - /1987/). Μ3 ΚΟΜΠΟΗΘΗΤΟΒ, <ίΤΟ KSCcīeTCS fleŽCTBHH, CaMHM IļeHHbllvi HBJĪfl-0TCH MMeroilļl'IH Η3υΜρ0Τβ./ΙΰΗ7:Ό MMMyHOno^as.ΊΗίΟΙίΰ'·»} aKTMBKOCTB U.HK-JIOCnopHH A.
IlHKJIOCnopHH SbiJI H3BSCT6H CKa^aJia KaK aHTHČMOTMK CO CJiaSblM προτΗΒΟΓρκδκοΒΗΜ seftcTBueM, ero 3HaqHTejii>Hoe WM>/iyHonoflaBJiHiomee fležcTBue ābiJio otkpmto jwnn> nosjļnee' (J. F. Borei h flp.: Immunology 32, 1017 /1977/). ΠρΗ noMoiiui utejroiī cepnH. οπητοβ Ha jkhbom opramraMe h b npo-6ίφΚ0 MOJKHO 5HJIO flOKa3aTL·, tJTO ΙίΗΚΛΟΟΠΟρΗΗ A, C H G HBJIHIOTCH Sojiee cneuH$nqecKHMH θκτηβηημη BeinecTBaMH, qeM n3BecTHHe so chx nop nMMyHonoflaBJiHK)mMe JieKapcTBeHHhie cpeflCTBa. 0Ka3ajioeb, ήό LiMKJiocnopHH A noflaBJiHeT KaK ryMopajn>Hb$, τβκ m iļejunanHpHbift HMMyHHbIŽ 0TB6T. ΠρΗ HCCJieaOBaHPM MeXaHH3Ma fleHCTBHH dblJIO yCTa-HOBJieHO, qTO oh nojiaBJiHeT npojm$epaunio Τ-κϋβτοκ h npepbmaeT CHHT03 HHTepJieMKMHa-2. 0 ΠρΗΜθΗΘΗΗΗ LIMKJIOCnopHHa A B Ka^eCTBe JieKapCTBeHHOrO cpeacTBa ripu nepeca^Ke opraHOB y qe.noBeKa cooomajm b 1978. ΠβρΒΟΘ ΠρΗΜΘΗΘΗΜΘ 6biJIO OIĪMCaHO B CBH3H C IiepecaflKOM ΠΟΗΚΗ (R. J. Calne h ap.: Lancet 1978/2, 1323) h TpaHcnjiaHTaiiMeH koctho-ro Mosra (R. L. Powles h ap.: Lancet 1978/2, 1327). Πρημθηθηηθμ BiMKJiocnopHHa A npn nepecaflKax opraHOB mojkho npenaTCTBOBaTB ot-TopjKeHHio opraHa (no^Ka, nofl}KejiyfloqHaH 2Kejie3a, πθ^θηβ,·. cepfliļe, jierKoe), h TpancnjiaHTaiiHHX κοοτηογο M03ra ottopjkbhhio κοοτηογο ΜΟ3Γ0. Π/ikjiocπορΗΗ A c ycnexoM npHMeHHjm Tarace ajīh Jienenus onpe-flejieHHbix ayTOHMyKHhix saāojieBaHMM (HanpHMep, BocnajieHHe cocy- - 3 - LV 10500 AHCToro TpaKTa rjia3a, peBMaTimecKM βρτρκτ, 'iem'fMaTUfi mmafi n THJKe.naH MMacTeHHfl). Πο naTeHTHoĀ JH*TepaType sjih nojiyneHHK coflep»amHx κομπ-JteKCHblfi UHKJIOCnopHH φβρΜδΗΤβΙίΗΟΗΗΗΧ O'fJlbOHOB πρΜΜβΗΗΛΜ flO CHX nop KyJīr>Typbi cjieflyfoiiĶix iiiTaMMOB MXKpoopranx3mob: Cylindooerpon luci duni Booth. , NRRL 5760 (naTeHT lilBečiiiapMH N 589 716), Tolvpocladium inflatum Ganīs, NRRL 8044/ no dojiee CTapon ho-MeHKJiaType: Triclodarme polysporum (Link ex Pērs.) Rifai, na-τθητ liiBenuapnM N 603 790 no Bissett (1983) Tolynflatunw, Ganīs, 1971 HBJifleTCH chhohhmom c Tolyniveum (Rostrup) Bisset. comb. nov. h Fusarium Saleni, MC1-1549, 1-401-1550 (onydJiHKOBaHHas 3a-HBKa na naTeKT Ηποκμμ N 62 63093) . OflHaKo npoMunuieHHoe mnojib-aoBaHMe θτμχ iiiTaMMOB MMKpoopraHH3MOB b enocodax (±)epMeHTaiiHH HMeeT tot HesocTaTOK, ^to BpeMeHa <|sepMeHTauHJī npoflOJDfoiTejiBHbie (11-14 flHež), nojiynajoT Majio κομπλθκοηογο UMKJiocnopHKa (okojio 20-30 ΜΓ/MJl) μ ΚΟαφφΗΙίΗΘΗΤ Π0Λβ3Η0Γ0 fleHCTBMH BblfleJieHMH ΚΟΜΠ-jieKca η3 $epMeHTaimoHHoro dyjiī>0Ha hm3khh (okojio 40%). B xofle HccjieflOBaHHM hcKajin ιιπήμμη MviKpoopraHH3MOB, npn noMoinn κοτορΒίχ mo^kho noJiyyaT& KOMnjieKCHbin βητμοηοτηκ unKJiocno-PHH MJIH ero ΚΟΜΠΟΗΘΗΤΗ B BbICOKOii KOHUOHTpaUHH Η πρκ āOJiee' Bbl-roflKbix ycjūOBJisx, yeM flo chx nop. B npouecce cepHHHbK Hccjieflo-BaHHž, KOTopbie pacnpocTpaHHJiMCb Ha μηογο Tbicsq mTaMMOB rpw6Ka, cmotjih BbiopaTb npHHaflJietfaiitKiī κ Bimy Tolypocladium MHKpoopra-hmbm, KOTopbiiī 3a dojiee κοροτκοβ BpeM» h b dojiee SHa^iTejibHOM KOJiM^ecTBe dM0CMHTe3iipyeT κομπλθκοηβιη iļHKjrocnopnH, neM η3βθοτ-Hbie I1IT3.MMbl, M KOTOpbIM, KaK HOKasaJIM ΤΘΚΟΟΚΟΜΗ^ΙΘΟΚΚβ HdCJieflOBa- hhk, He MfleKTnneH hh c οληημ H3 ηββθοτηηχ npoflymīpyjomjwx ijmk- - 4 - JIOCnopMH IIITaMMOB ΓρΗΟΚΒ. HOBbIM ΙΙΙΤβΜΜ (07/93) - KOTOpbIH ΠΟ co6ctb0hhhm nccjieflOBaHMHM cjieziyeT paccMaTpMBaTB κβκ hobo-ro oTJUi^HTejiBHoro TaKcoHa po^a Tolypocladium - 6biJi onnoaH nofl Ha3BaHweM Tolypocladium varium specias nova κβκ γολοτμπηηΛ ΙΙΙΤΒΜΜ.
MsočpeTeHHe ocHOBbiBaeTCH, c.ne,noBaTeJiī>HO, Ha tom οτκρβιτιιη, B pe3yJIBTaTe φβρΜΘΗΤβΙίΜΜ ΗΘΜ3Ββ0ΤΗ0Γ0 flO CHX nop IIITaMMa THna Tolypocladium varium specias nova (flOManiHee HaaBaHHe: CV/ 93), κοτορΜΜ 5biJi flenoHHposaH īiojļ HOMepoM NCAIM( P)F 001005 b EyflaneniTCKOM Υηηβθρομτθτθ caflOBOflCTBa η πηπϊββοζ προμηιμθηηοοτμ B HaLtHOHaJIBHOM COČpaHMH CeJIbCK0X03HHCTBeHHbIX H ΠρΟΜΜΜβΗΗΗΧ MHKp00praHH3M0B, M03IIH0 nOJiyqaTL· $epM0HTaH,MOHHbM 6yJIi)OH C O^eHB BHCOKOM ΚΟΗυ,βΗΤρΒΙΙ,ΗβΜ PMKJIOCnopHHa. ΦθpMeHTaiļHOHHblM 6yjn>OH Ha-pfffly C OCHOBHbIM npOffiTKTOM liMKJIOCnopHHOM A COflepJKHT B ΗβδΟΛΒΠΜΧ. KQJM*ieCTBaX IļMKJIOCnOpMH B M IļMKJIOCIIOpMH C.
HcnOJIL3yeMbie JļJlH CMCTeMaTH3aHMH ΠρΗ3ΗΒΚΜ BbffleJieHHOrO HO-Boro niTaMMa rpnoKa cpaBHHBaioT hhm© c ββ^κηθηκιμμη ms flMarHOCTM-qecKKX xapaKTepHCTWK μββθοτηηχ βη,ςτοβ Tolypocladium.
Poā Tolypocladium 6bui orriicaH b 1971 Gans κβκ HOBbift BHfl KacejisK)iipiX no^By Ivfoni 1 ialas. Ero xaparcrepHHMH cbohctbbmm hbjih-iotch: MefiJieHHbiH ροοτ, nopyiiiKooopasHbie oejibie hjīh SejiOBaTbie oaK-TepnajīBHhie kojiohmh, κοηθ^ηηθ n Ροκοβηθ, pacnojuoaeHHbie ^bctm^ho Hcl KOpOTΚΗΧ GūKOBbIX ΒΘΤΒΗΧ φ'ΑεϋΙίΊ^Η C CJiJTBHO pa3čyXIHHM OCHOBaHH-6M, Ka KOTOpOM MOJKHO pa3JHWHTL· HMTeBiiflHyiO, ^BCTO flyroo6pa3Hyio HaKJioHHjno mežKy, KOTopas oKaHUMBaeTca μβχθκβκομ οΛΗοκΛβτο^ΗοΛ KOHHflMež (γολοβκοζ). BHyTpH 3Τ0Γ0 posa Gans pa3JUīwajr 3 cjreffino-ipx HOBbix BKfla: T. geodes, T. cylindrosporum η T. inflatum LV 10500 T. geodes ΗΜββτ nopasMTeJibHbiM, HanoMHHaiomn# 06 aKTMH0MHiļ6Tax 3anax seMJin, κοτορυΛ nojiKocTLio OTcyTCTByeT y ηοβογο T. varium sp. nov. Cy/93. KpoMe τογο, κλθτκμ hocht6Jih T. geodes sHa^HTeJib-HO yjift3, 40 M KJĪ8TKH HOCMT6JIH T. VariUJĪl Sp. HOV. 07/93. KOHMflMM T. cylindr oosporum xapaKTepno λληηκηθ h pacTSHyrtie, κοη^η^ριη T. varium sp. nov. 07/93, HanpoTMB, mapoBMflHfaie hjih cjiado pacTS-HyTbie. ΚοΗ,ςΗκ T. inflatum BJiJimiTmecKm h RJiMHHee, yeM komņm T. varium sp. nov. 07/93. §mmm T. inflatum pacnojmeHbi mw He-nocpeflCTBeHHo Ha Γκφθ suut Ha KJieTKax flep^arejin rmhož 2-3 m KJiySKOM. fljiH ffiMajīH^oB T. varium sp. nov. cy/93 pacnojioifieHMe KJiydKOM He HBJĪHēTCH xapaKTepHbIM H HauJiiOflaeTCff TOJIBKO B OTfleJIBHbIX CJiy-qaHX B6PTMKaJIBHbie KOJIOHMH Bcex ΗΒΒβΟΤΗΗΧ RO CHX ΠΟρ BHflOB Tolypooladium oeūHe, <jto oāī>HCHHeTCH} b nepByio oqepefl£>, āejibiM, BaTOodpasHBiM bobapihhm MMuejmeM. HHūuiHS CTopona daKTepHaJLbHHX kojtohhh BSJīTOBaTO-cepaa, oecuBeTHaa mjih KejiTas, xapaKTepHbiž PaCTBOpMMbM ΠΗΓΜΘΗΤ Ηβ npOBBBOflHTCa. Jf B 3T0M ΟΤΗΟΙΠΘΗΗΗ T. varium sp. nov. 07/93 oTJnraaeTCH οτ Μ3Β8οτηβιχ bh^ob: Ka nHTa-TejiBHbix cpesax c rjnoK030Ā u πθπτοηομ, Tara® Ha iiMTaTeJiLHbDC cpeflax C COJIOflOBhIM SKCTpaKTOM/flpOSOK0BbIM SKCTpaKTOM ΓρΗδΟΚ BH-pafiaTUBaeT τθμηημ, BapbHpyeMb$ bo ΒρβΜβκκ οτ rycToro ceporo flo ^epHoro iiseTa, πκγμθκτ.
ĪIpH CMCTeMaTPmeCKOM CpaBH0HHH ΗΟΒΟΓΟ UITaMLMa C K3BeCTHbJMH BHRaMH Tolypocladium čbuia ncnojn>30BaHa npejKBe Bcero cjieayTūinasī JiHTepaTypa: Ganīs, W: Persoonia 6, 185-191 /1971/ ) - Barroji, G. L. Gan. J. Bot. 5R, 439-442 /1980/ m Bissett, J.: Can. J. But 61, 1311-1329 /1982/. - 6 - #MarHOCTM*iecKMe πρμβηβκη Tolypocladium varium sp. nov. CY /93 μο;ληο odočmHTb cjieflyK)iii^M očpasoM. EaKTepMaJiLHbie kojiohhm B03pacTOM 10 nuePi meicm fluaMeTp 10-25 mm, ηχ noBepxHOctb ποκ-pb!Ta BSTOOOpaSHbLM SeJIbiM, HaCbHHeHHblM CnOpaMH B03flyniHbIM miļem-eM. Hhjkhhh CTopoHa 6aKTepMajiBHbix kojiohmm «ejiTOBaTo- cepas mn rpH3H0-cepas. Ha μηογμχ κομπλθκοηηχ nHTaTejii>Hhix cpe^ax odpa3y-6TCH TeMHO-CepHŽ paCTBOpMMblft ΠΗΓΜ6ΗΤ. Ha ΓΜφβΧ B03flyiIlH0r0 MM-uejiMH cnopooGpasoBaHHe npOTeKaeT ksk κοηθ^ηοθ, TaK μ āoKOBoe. ®iajiHflbi pacnojiciJKeHhi MecTaMK hjīh MHorfla TaiUKe nyuKOM Ha Γΐιφβχ MJIM Ha κοροτκκχ ( ajihhom'2-3 m) KJieTKax flepjftaTeJiH. ΦΗΖΐΜJļbl coctoht M3 pa36yxfflež ochobhoh uacra (2-4 x 2-3 mm) h RJimoft HMTeBHflHOH KpOMKH (2-4 X 0,5-0,6 M). KOHHflHH TOJIOBOK He60JH>- nwe (2-3 x 1,3-2,2 m) , b odmeM, niapoBHflHbie h poBHbie. BHflOBoe onpeaeJieHHe "varium” othochtch k η3μθη^ηεοοτη Φηθληηοβ. Tol. varium npoHBJiseTCH b CBoeM Λ®ΐ3ΗβΗΗ0Μ iļMKJie onpeflejieHHy}o exo-jfoscTB c Ganusharposporium, ο ηθμ no3»e ems dyfleT roBopHTBCS. IifraMM cy/93 He nenojn>3yeT pa<$MH03y, oflHaKo xopomo pacTeT b npHcyTCTBHM hctohhmkob yrjiepofla: MaHHHT, MH03HT, caxapo3a, $pyKT03a, paMH03a, raJiaKT03a, rjiKK03a, apa0MH03a h Kcmi03a. Ha nHTaTejiBHOM arape h cojio^obom arape ero Kyjiī>Typbi pa3BHBaioTCH cjiado, Ha οοθβομ arape, arape C apeK h KapTotJejiBHOM arape ohh pacTyT ycme.HHo. OBcsHO-flpojfCKeBOii SKCTpaKT, rjnoK030-flpoaoKeBoii SKCTpaKT H OBCHHhlM arap He CJIHUIKOM ΠΟβΧΟβΗΤ RJŪI COXpaHeHHH. LV 10500
TaejiHua 1 f- Tolvpocladjum varjum CY/93 e 3. 3 M **-( O "D 0.0 cn cn Γ' r-H o · U Μ Γ-Ο Ό .-I o. c r-~ ]>ι *<—v σ >>čo ι— u o ε Ē 3 3 U • rn o n ac cn (Γ. r i—1 o . U tJ c ο ό ία c r « HH1T o >. a 1— u c E 3 3 Ό ~ cn • f—H 3 U η o ir. - cx (U >^~o 3 H Ot : o ai c 3 1— PIC e c 3 r> ·<—1 ο n <n cn iN i—1 Γ~ - u N o cr -i a gj >.n r f-i a r o ai 1— π cn ε ε —i c-l 3 <3· aj c-i vj Ό Ο λο o an .c cn o >u •'"i 1—1 CJ U O (-i— n <r c a a u cn o U \0 O J--£-< Ē ΙΛ Ό 3 c a _j ,r“» ·<-» Q£ ' H uo; O»' 3 2: f \ _| pa3Jio«eHHe ueji.ruo.no3t I, _ _ - - - 4 44 4-44 BO 3J3yn3HbIH MHUeJIJIHH 3eJieHbIH HJIH r0Jiy60BČ το3βτι. 1 - - - 4 + - 4 (-4 : KpaCHblH paCTBOpHMblH ΠΗΓΜβΗΤ - - - - - - 444 :;KOpHMHeBbIH ΠΗΓΜβΗΤ Ha jKejie3onenTOHOBOM 'arape - " - - - - +++ ΤβΜΗΟ-ΧΟρΗΗΗ.ΠΗΓΜβΗΤ Ha chht .TjiHtiepHH-ara pe - - - - - ,4 44 - ycBaeMocTb caxapo3bi 444 +++ +++ 4 44 + -T _ 444 ycBaeMocT& HHyjTHHa - - - - - 444 ροβτ npn 5°C 444 444 +++ 444 + m ± - * ροοτ npn 37°C - - - - · - 4 4 44 CTOKKOCTb K 60-mhh. HarpeBy πρκ 50°C ++ + - - «* - +++ ļ +++ nojiyHeHHe KHCJiOTbi : H3- rJII0KO3bI +++ l’ 444' ' + 4 H---l· 4 4*4* +++ ycBaeMocTb Ma-casm-UHJiaTa - - - - +++ t 444 ycBaHeMOCT& ή/a-Majio-Hara + + ++ ++ - - ++ ++ ycBaHeMOCTb //a-TapT-paTa + ++. ++ - - ++ ++ JIHJTOBblfi paCTBOpHMblfl ΠΗΓΜβΗΤ Ha THP03HH- -JieiiHHH-arape - - - +++ + + + - - 5760 - 9 -
Tol. varium pe3K0 OTJBiuaeTCH ot Tol. Trigonosporum, TaK κ.ακ nocjieflHHH npoflyu,MpyeT Trigonale conidia, κοτορπΛ ημθθτ cjiaoo H3orHyTHe Kpaa, Tol. varium OTjmaeTCH Tarase ot Tol. balanoides, TaK KaK nocjieflHHft npoflyii»pyeT οοκοβηθ ΚρΟΜβ ΤΟΓΟ, OTJIHuaeTCH rOJKjTMIIHfcJM SITclMM (Cy/93) Tolypacladium varium n. sp. ot 3aperHCTpwpoBaHHoro τηποβογο ΐίίΤαΜΜα (CBS 714. 70) T. niflatum Taranē τθμ, hto nocJieflHMH o6pa-3yeT Ka cojīosobom arape οομλβηκιη oejibiM B03flymHhM MnpejumiS, b npoTHBonojiojKHocTb 3TOMy T. variumm sp. nov. cy/93 He oāpa3yeT ηκκβκογο miļeJim. TarcoBož ίιιτβμμ T. niflatum npon3BOflHT b cy6c-TpaTe MMuejuiHH Ha JKejieso-nenTOHOBOM arape KpacHOBaTO-KopnHHe-BbīM KOKe^HHM ΠΜΓΜΘΗΤ. B npoTHBonojio«HocTi> 3T0My MHpe.MM#-cy5cTpaT T. varium sp. nov. 07/93 ocTaeTCH 6jteflHO-jfiBJiTbiM. T. varium sp. nov. cy/93 ηθ nepepafiaTHBaeT apa6MH03y, b npoTMBonojroxHOCTB T. niflatum. T. niflatum saeT c HPiTpaTOM ΗΕτρκκ οομ.πκηημ miļejumft, b npoTHBonojiojKHOcTh 3TOMy T. varium sp. nov. cy/93 eflBa pacieT.
CpaBHeHMe c saperHCTpMpoBaHHbiMH ιιιτβμμβμη fflpyr«x bmhob Tolypocladi urna, a Tarae co niTaMMaMH Trichoderma polysporum ATCC 16641 h cylindrocarpon lucidum NRRL 5760 npHBe^eno b Tač-JiMue 1. IīpeflMeTOM HsoopeTenHH HBJiHeTcs cnoeoā nojiyyeHMH κομπ-JieKCHOrO aHTMCiMOTMKa IļHKJIOCnopHHa H/MJIM ero KOMIIOHeHTOB: UHK-JĪC-CnOpHHa A, UHKJIOCnopHHa B H UHKJIOCnopHHa C KyJILTHBHpoBaHMeM 5hocMKTesHpyroiij,ero KassaKKHe aKTHOHOTHKM m’TaMMa rpHOKa b coflep-*ame# ycBanEaeMbie mctohhhkh yrjiepofla h a30Ta h MHHepajīBHbie co- LV 10500 - Β· - m πΜΤ9.τβϋί>Η0Η cpefle, πρπ aapo6Hbix ycjioBHHX, h OTflejieHHeM οδ-pa30BaHHbix npoflyKTOB. flira cnocoāa οτλϊ'Γιητθ.πβηο# ^βρτοζ HBJiHeTCH to, qTO npoayju;Hpy}omHM κομπλθκοηηιζ 3βτη6ηοτρικ uhkjīoc-πορηη ιΐίΤΕΜΜ ηοβογο BHfla rpHdrca Tolypocladium varium, npeflnoq-thtojibho flenoHMpoBaHHbiž nofl HOMepoM NCAIM(P)F 001005 b Eyfla-neniTCKOM Υηηβθροητθτθ eafiOBOflCTBa μ πμπϊθβοΛ προΜΜίυιβΗΗοοτίί b HaUJHOHaJiBHOM COOpaHMM CejrBCKOX03HMCTBeHHbIX M IipOMb!IIIJieHHbIX MHK-poopraHM3MOB uiTaMM Tolypocladium varium sp. nov. C7/93, KyjibTHBMpyioT b coflepffiametf ηοτο^ηηκμ yrjiepofla, a Taranē opraHH-qecKMe h HeopraHimecraie mcto^hmkm asoTa κ MMHepajiLHbie cojw m-TaTejibHož cpese, ripu aapo6Hbix ycjiOBHHX, npn TewnepaType 25-30 C, b cjiy^ae ηθοΟχοαμμοοτη BHflejiHioT 06pa3OBaHHbiž κομπλθκοημΑ aHTHČHOTHK mm ero κομποηθητη hs <#epMeHTaiļMOHHoro 5yjiBOHa μ b cjiy^ae HeočxoflWMocTH o^HinaiOT. ITO ΗΒΟδρβΤβΗΗΙΟ flJĪH ΠΟΛ^ΘΗΗΗ ΚΟΜίυΐβΚΟΗΟΓΟ aHTHOHOTHKa UHKJIOCnopMHa ΙφΗΜΘΗΗΙΟΤ npefllIOtlTHTeJIbHO KyjIbTypy ΗΟΒΟΓΟ EUTaMMa Tolypocladium varium sp. nov. 07/93. 3tot inTaMM BCJieflCTBHe cbo-ero ābicTporo pocia HBJiHeTcs ocoāeHHo BbirofiHbiM. Caxapo3y, rjiro-K03y, cop6o3y, Majn>T03yJ $pyKTOsy, KpaxMaji, rjomepHH h pa3JiHy-Hbie jKHpu h Macjia oh χοροπιο Hcnoju>3yeT k&k mcto^ihhk yrjiepofla. B Ka^ecTBe ncTo^HMKa asoTa npMHHMaroT b pac^er pasjiH^Hbie opra-ημηβοκμθ HCTOtfflMKH a30Ta, HanpMMep, BOfly Ha6yxaHHH KyKypy3bi, Π8ΠΤΟΗ, flpOJfOiieBO/I SKCTpaKT, MHCHOŽ SftCTpaKT, KHTpaT HaTpuH, HHTpaT aMMOHHH, CyūLCļ)aT aMMOHHH M paSJĪM^Kbie aMJĪHOKHCJIOTbl. Kpo-ΜΘ 3thx hcto^khkob yr.nepofla m asoTa nHTaTejīBHas cpe^a mojkbt coflepjuāTb eme MMHepajībHbie cojih nanpHMep, ΧΛορΗΟΤίώ K3jīmm, cyjii>(|)aT μβγημη ma mcjmA φοοφορΗοκκοΛΗίί Ksuim), μηκροθλβμθητη (HanpHMep, MeflL, MapraHen; h atejieso), a Tarace βητρμηηη η πθηο-racsiiĶie BeinecTBa. ĪTo npejB[nowTHTejibHOMy BapnaHTy ocyiipcTBJieHHH cnoeoča no Η3θ6ρβΤβΗΗΚ) OT ΑβΠΟΚΜρΟΒβΗΗΟΓΟ IlOfl HOMepOM MCAlM(P)F 001005 B ByflaneiDTCKOM ΥΗΗΒθροητβτβ ca,zīOBOflCTBa η πμιπ,θβομ προμβιιμθηηοοτμ B HaUHOHaJLbHOM COČpaHHH Cejn>CK0X03SĀCTBeHHbIX H ΠρΟΜΗΙΜβΗΗΗΧ MHKpoopraHMSMOB fflTaMa Tolypocladium varium sp. nov. 07/93 Remm noces KyjiBTypbi Ha kocom arape, cycneH3HeĀ cnop n cycneH3Hefl MHIiēJIHH ΚΟΤΟΡΟΓΟ ΠρΗΒΗΒδΙΟΤ ΠΗΤαΤθϋΒΗΤΚ) Cpefly HHOKyJIHTa, 3aT6M So5aBJisK3T Kyjii)Typy HHOKyjiHTa B03pacT0M 3 rw k npnSjiHsiiTejtbHO flec'HTKKpaTHOMy KOJuraecTBy epesbi npoflyu;HpoBaHHS h HHKy6npyioT cpeny npoflyiņipoBaHHH npn 25-30 C, npeflnonTHTejiBHO 25’ C, b Te-qeHHe 5-7 jļnevi. Bo BpeMH (fepMeHTaiņui no^epatsiBaioT BejnniHHy pH MejKfly 6,0-2,5, πρ6Λποητητβ.3ΐ)Ηθ πρκ 5,2. ΦβρΜβΗτβφΐΗ προηοχο,ςΗΤ npH a3p00nbix '/CJIOBHH'Ā Η ΓφΗ ΗΗΤΘΗΟΗΒΗΟΜ ΠθρβΜβΙΜΒΕΗΗΜ (950-1000 mhh ). ΠοτρβδΗοοτί. Kyjn>Typu b bossyxe cocTaBJiHeT οκο-jio 300 Ji/nac.
Bo BpeMH φβρΜΘΗΤΒΠ^Μ 3a COflepjSaHHeM aKTHBHOrO BenļeCTBa B φΘρΜβΗτειίίΗοκκοΜ 0yjo>oHe HaāJiiOflaiOT nyreM MHKpooMOJiornnecKoro HSMepeHMH BeJIM^MH H nyTeM atHflKOCTHOft Χρ0Μ3Τ0Γρ3φΜΗ npH BbICOKOM itaBJieHHn (HPLC). Kor^a coaep«aHne aKTMBHoro seiuecTBa nocīnrjio CBoero MaKCHMyMa, φερΜθΗτβΐίΜΐο npeKpamarar w aHTn6noTMK BbiflejnnoT μθτοηβμμ 3KCTpaKiņiM h/mjih aflcop6i[Mn.
Aspargillus japani cus n MMKpoāHOJiornnecKoro HSMepeHira βθηθ^ηη ncnojn>3yioT OKa-3aHH0e nHKJIOCnopnHOM A Ha ΜΗΟΓΜβ ΓΜφθΜΗΙ1ΘTbl H36MpareJH>H0e τορ-MOJKBHMe pocTa. B KanecTBe TecT-opraHH3MOB Hanāojiee ποαχοαηιπ,μμμ HBJiHiOTCH Asparg i 11 as η 1 gerr, LV 10500 -//-
Curvularia Iunata (M. Dreyfuss h ap.: European J. Appl. Microobiol. 2, 125-133 /1976/). ftjifl aHaJuraa jfttiflKOCTeft xpoMa-
TorpatjMeii npn bhcokom saBJieHHH (HPLC) $epMeHTautnoHHoro 5yjubOHa ΓφΜΜΘΗΗίΟΤ φΜ.ΠΒΤρ3Τ pa30aBJieHHbIX B ΟΤΗΟΠΙΘΗΜΜ 1:10 MeTaHOJIOM npoo (annapaT: ISRB, MOfleji£> Hacoca 2150, Mo^eJiB yjiBTpa$iiOJieTO-Boro aeTeKTopa 2151; H3MepeHMe npn 220 hm; kojiohkb B5T -10C-C8, 7 mm; BbicoTa kojiohkh 25 cm, BHyTpeHHHž flnaMeTp 0,4 CM; MIRKm 3ΛΘΜβΗΤ: aiļeTOHMTpMJE Η TpHSTHJiaMHH φΟΟφορΗΟΗ KHCJIO-th pH 6,8) η B OTHomeHHM 65:35). . 3ΚΟΠβρΗΜ6ΗΤ2ϋΠ>ΗΗβ ΟΙΊΗΤΗ nOKB3aJIH, *JTO HOBBI& IffTaMM Tolypocladium varium sp. nov. cy/93 fiae-ϊ c oneHB bhcokhm bhxo- flOM (950 Mr/MJl) KGMHJieKCHblH aHTHOHOTHK, COfiepmUīHM B Ka^eCTBe ΟΟΗΟΒΗΟΓΟ npOflVKTa IiMKJIOCnopHH A H h’apfffly c KMM Β ΚΒΉβΟΤΒΘ Ηθ" dQJILII!riX ΚΟΜΠΟΗΘΗΤΟΒ UHKJIOCIIOpIĪH B H UMKJJOCnOpHH C.
BbIfle.HeHWe ΚΟΜΠΛβΚΟΗΟΓΟ ^ĪKJIOCīTOpMHa M3 φβρΜβΗΤ3Ι1ΜΟΗΗΟΓΟ 6yjii)OHa uejiecoodpasHo npoBOflHTb sKCTpaKiiMeft. BbiroflHO nepea sk-CTparaņieū ysejiHTL· mmi^-mM m πητΒτβϋΒΗΌΜ cpeflbi Φμβτροββημθμ HJIH IjeHTpHlferrHpOBaHHeM. 0t OTfleJieHHOŽ ČHOMaCCbF ΠρΗϋΗΠΙΠΗΜ aHTH-Ghothk liejiecoočpasHo npoMUBaTb ηηβιπμμη cimpTaMH, ocodeHHO Me-TaHOJioM vijai KBTOHaMM, ocodeHHO aiļeTOHOM, b to BpeMH KaK fljuī aKCTpaKUHM paCTBOpeHHbK Β ΠΗΤβΤβΛΒΗΟίί Cpefle UHKJIOCnopHHOB Ha-XGĀHT ΠΡΗΜΘΗΘΗΗΘ Hc C Μθ ίίΜΒ αβMbic C BOflOft paCTBOpHTeJĪH, HanpHMep, 3THJiaueTaT, n-oyTMJiaiļeTaT, ī-6yTiiJiaueTaT, αηχλορμθτθη, 1,2-flHXJīop3TaK Km χϋοροφορΜ, npeĀnoqTHTejii)Ho n-6yTHJiaii;eTaT. HojiyqeKHbiH aKCTpaKi^reH ebipotf προ^νκτ 3 ar pa 3 ne h npo2yiinpoBaHHbi-
MH rpnāKOM KpaCHbLMM M JT/IJĪOBbiMM ΠΗΓΜβΗΤ3ΜΗ. IIOCJieflHMe MOJKHO yflajiHT& a^cop6iļMePi na aKTMBHOM yrjie mm cMJinKarejie. MMefombiecH - /1 - npn cjiy^ae β φβρΜΘΗτβρΗΟΗΗΟΜ 6yjibOHe npMMecH jumMflHoro xapaK-Tepa (HanpHMep, aHTMBcrreHHBaTeJiM) mojkho yflajiHTi> flHcneprMpoBa-HMew cbiporo npojiyKTa b neTpojiežHOM a$Mpe h coflepjKanņiM 10% BOflhi MeTaHOJie. jlHnHji;H006pa3nbie πρημθοη nepexo,qHT b $asy neTpojietfHO-ro ac^ipa. Jis BOflHOMeTaHOJioBoft φδ3Η yqajiHiOT BbinapMBaHHSM b Ba-KyyMe, οοτβηχιφποοη BO,zi;Hyfo φa3y 3KCTparwpyioT flnxjiopMeTaHOM m bh-napMBaiOT SKCTpaKT. Tia nojiyqeHHoro TaKMM nyTeM ompHHoro npoflyKTa μομηο οταθπητβ ρκκϋοοπορΗκ A m ,npyrHe očpasoBaHHbie b MeHbfflēM KOJIH^eCTBe ΚΟΜΠΟΗβΗΤΗ UHKJIOCnopHKa Χρ0Μ3Τ0Γρ3φΗβίί; Ha KOJioHKe mjih nepeKpHCTajumsapiieM. Πρκ cpaBHeKMM c M3BecTKbLMH flo cmx nop cnoeoSaMH cnocoā no Η3θδρβΤβΗΗ.Ό ΗΜΘΘΤ pfifl ΠρθΗΜ'/ίΠ,βΟΤΒ, KOTGpbie MOJKHO C^MMPĪpOBaTb cjieflyioiipiM oopasoM:
1) OcHOBHoe npeHMyiii,ecTBO cnocoča coctoht b tom, hto DiTāMM
Tolypocladium varium sp. nov. cy/93 flaeT b HecKOJiLKo pas 6ojn>- ·· rnee κμμηθοτβο κομπλοκοηογο UHKjrocnopHHa, nojiy^aeMoro M3BecT-HHMH flO CHX πορ ΜΘΤΟ^ΜΜ, ΙφΜ JiadopaTOpHbIX yCJIGBMHX KOJIHHeCTBa flocTHraioT ro 950 mt/mjl 2) HOBbIM IDTaMM flaeT KOMIUfeKCHHft pHKJIOCnOpMH ČJiaronpHHTHO-ro cocTaBa ( okojio 85% KOMnjieKca coctoht H3 pHKJiocnopHHa Δ), h n03T0My $6pMSKTaiiMOHHblM 0yJIi>OH MOJKHO ΠρΟΰξδ Η ΘΚΟΗΟΜΜΗΗΘΘ ΠβρΘ~ paoaTUBaTb, ηβΜ θτο HMeeT mccto npn M3BecTHhix cnocodax.
BbiflejieHHbie npoflyKTbi ηΑβΗΤΜφΜυ;ΜροΒ3Ηη HCCJieflOBaHMHMM yjo>-τρ3φκοηθτοβογο cneKTpa, HH$paKpacHoro cnerapa, cneKTpa ΙΗ-HflepHoro MarHMTHoro pe30HaHca κ 13C-HflepHoro μειγηητηογο pe-30HaHca, aHajiH30M aMHHOKMCJioT h 'mRKOCTHoPi xpoM3Torρ3φηθM npn BbiCOKOM flaBJieHMH. LV 10500 - /3. - jfeočpeTeHne ποηοηηκτ nospoāHee Ha ocHosaHMH cjieflyioinwx npMMepoB. ΙΊΡΜΜΕΡ 1: līs BbipaiiļēHKOH Ha cosepmHipM cojio/ioBbiM SKCTpaKT h apojote-boh aKCTpaKT kocom arape BospacTOM 9-7 j\hsm KyjibTypbi uiTaMMa 7olypocladium varium sp.no v. 07/93 (NCAIM (P) F 001005) īipn-TOTOBJIHfOT c 5 mji 0,9 %-hopo paCTBOpa ΧΛΟΡΜΟΤΟΓΟ HaTpHH cycneH-3HI0 Cnop. ĪTpH nOMOIHH 1 MJI 3T0M CyCn0H3HM ΟΠΟρ 3α0ΘΒ3ΐ0Τ 100 MJI nHTaTeJttHOŽ cpeflbl C MHQKyJIHTOM, KOTOpaH HaXOflMTCH B KOJIČe 3p-JieHMežepa oobeMOM 500 mji. .
CocTas nHTaTejiLHOH cpejļbi c HHOK.yjihtom 1C rjiiOKOsa 40 r Κ33ΘΗΗ0ΒΗίΐ ΠβΠΤΟΗ 5 Γ KHTpaT HaTpHH 3 r KMCJIHM $Oe$OpHOKHCJIbJM KaJIMM 2 Γ xjiopncTbič[ KajiHž 0,5 r cyjib$aT Mcithhh · 7H^0 0,5 r cyjiL(|)aT jKejie3a (11)-711^0 0,01 r B 1000 MJI BOflOnpOBOflHOft BOflbl rīMTaTēJibHyio cpesy ycTaHaBJiHBajOT so pH 5,2 m nocjie θτογο GTepMjiM3yH3T πρκ 121° C b τθηθηηθ 25 MHHyr.
KyjiBTypy BCTpHXHBaior Ha annapaTe rjih βοτρηχμβ3ημη (340 MHH, OTKJĪOH6HM0 3,5 CM) ΠρΜ 25° C B T0H0HH0 3 flHeft. IIpH ΠΟΜΟ#ί 5 mji nojiy«eKHoft KyjibTypw HHOK.yjiHTa saceBaiOT 15 kojič SpjieHMeže-pa OČB0MOM 500 MJI, B ΚΟΤΟΡΗΧ h'a>:OflHTCH no 100 MJI CTepHJIbHO# nMTaTejfbHOM cpejibi FC1.
CocTaB πκτβτβ,πΒΗΟ# cpeflbi c ηηοκυ,πητομ FC1
rjnoKosa 80 r TpMnKaSHH 40 r Mo^eBHHa 2 r CyJIL·φaT aMMOHHH 12 r HHTpaT HaTpHH 3 Γ KHCJIbJŽ φοσφορΗΟΚΜΟΛΗή KaJIMH 2 r XJIOpHCTbltf KaJIHH 0,5 r cyJIx5φaτ MarKHH · 7HA0 0,5 r cyjiB(|)aT atejiesa (II)· TEjfi 0,01 r B 1000 MJI BOflOnpOBOflHOii BOflbJ
IfiiTaTejii>Hy]0 cpefly ycTaHaBJTMBafOT ro pH 5,2 m nocjie 3Τ0Γ0 cτepmjimsyfOT ripu 121° C b τθ^θηη© 25 μμηυτ.
Kojiod! BCTpHXMBafOT Ha annapaTe zum βοτρηχμβοημη (340 mmh , οτκΛΟΗβΗΜβ 3,5 cm) npM 25°0. 3a Μ3μθηθηη6Μ coflepaaHMa aHTKdno-THKa b $epMeHTaiņīOKHOM 6yjihOHe HadjnoflaiOT πρπ noMoma ΜΗκροδΜΟ-λογηηθοκογο Me-TOfla flHφφy3Hκ b arape.
MeTOfl βΜφιφγοΜΗ b arape r.eh MMKpooHOJiormecKoro HSMepeHMH BeJIHHHH OCHOBHBaeTCH Ha ΠρΟΤΗΒΟΓρΜδΚΟΒΟΜ fletfCTBHH IŅiKJĪOCnOpHHa A. B KanecTBe TecT-opraHH3Ma πρκΜβΗΗΐοτ ASpergillus niģer. Ak-THBHOCTL 05pa3OBaKH0ro ΚΟΜΠΛΘΚΟΗΟΓΟ IļHKJIOCnopHHa MBMepHIOT OT-HocHTejiBHO UHKJiocnopnHa A-CTaHflapTa. CosepjKaHue 2-8 mt/mji umk-jrocnopHHa A, πρηγοτοβλθηηηθ c βο^ηβιμ chhptom CTaHflapTHbie pacTBophi ztaioT Ha co^epaainež αρολοκθβοη aKCTpaKT m rjnoK03y ara-poBotf epefle sohh topmojkshmh inaMeTpoM 16-25 mm. LV 10500 - !S - (fepMeHTaanio npoflOJDiiafOT 168 qacoB, ποτομ BbmejiHioT oāpa30-BaflHbM KOMIĪJieKCHblii aHTHČMOTMK H3 φβρΜβΗΤ3ΡΜ0ΗΗ0Γ0 6yjIbOHa.
Sfe oflHoro jMTpa $epMeHTauKOHHoro 6yjihOHa, κοτορω# ripu npeKpameHMH $epM©HTapnH coflepjKHT 400 mt/mji κομπλθκοηογο φΐκ-jrocnopHHa, nojiyyaioT aHTHČHOTHKM cjieflyfomnM o6pa30M.
KneTKM yflaJIfii0T φΜΛΒΤρΟΒΘϋΗβΜ 1Δ3 φθρΜβΗΤ3Ι1ΗΟΗΗΟΓΟ 6yjn>OHa M npHJHfllHIHe aHTHSMOTMKM OTMbIBaiOT 2x200 MJI MeTaHOJia. Jfe BOflHO-MēTaHOJIOBOM ΠρΟΜΗΒΟ^ΗΟΗ JKHflKOCTH yflaJIHKJT MeTaHOJI npH ΠΟΗΗΛίθΗ-HOM flaBJieHHH M 3KCTpaTHpyfOT nOJHyweHHbIŽ BOfiHblPl KOHIieHTpaT 2x50 MJT n-6yTyTJiaiieTciTa. 3fe ĢKJIbTpaTa φβρΜβΗΤΕφϊΟΗΗΟΓΟ 6yjH»OHa 3KC-TparHpyfOT KOMnjieKCHbiĀ uhkjiociiopmh 200 mji n-oyTHJiau;eTaTa. 3kc-TpaKTbi coeflHHaioT, cymaT Kāp; āesBOSHHM cyjtt$aTOM HaTpMK κ Bbma-pHBafor npw ποημκθηηομ flasjieHMM npyi 40° C. ĪIoJiyqeHKbiM cbipow npo,r;yKT (3,7 r) pacTEoparor b 10 mji MēTanojia, μ ληποΦμλβημθ npMMecH ysajiHiOT 3JuoHpoBaHneM c μθτβηολομ Ka cjioe rejifl M3 Sephedex LH-20 (BbicoTa 40 cm, /piaMeTp 2,5 cm). CofiepjKaīpe KOΜΠJieKCHblM IļHKJIOCnopiiH φρβΚφΙΙί BbinapHBafC'T IipH IIOHHSteHHOM flaB-JieKMM npH 4Q°C. IlojiyqeKHbiH κομπJiekckhh LļHKJiocnopHH pa3fleJiHfor Ha ero κοΜποΗβΗΤΗ Ka πρηγοτοβλθηηοΛ h3 20 r eMJiMKarejiH (cmjm-Karejip 40, PeaHaJī, ByflaneniT) KuJioKKe SJUOMpGBaHHeM c noBbimaioiiiKM rpaflHeHTH ΧΛοροφορΜΗ-MeTaHOJia cosepaaHHeM MeTaHOJia. OjnoHpoEa-Hwe HauwHafOT c 100 mji χαιοροφορι»©. m npoflOJKuaior c 1.00 mJi - kojm-
VieCTBaMM aJIiOeHTa, B KOTOpOM CO^epKaKMē MeTaHOJia npH KaKflOM CTa^HM noBbimaiOT Ha 0,5%. 3a coflepataHHeM UHKJiocnopHHa b xpoMa-τοΓρβφΗ^βοκκχ φρΕκρκΗχ cjieflHT npH πομοιιιη tohkocjiohhoh xpoMa-τοΓρβφκκ (iuiacTMHbi c cMJMKarejieM 50F254, DC ajnoMMHneBaa $ojib- ra, MepK; KOHCTpyKTOp: 3THJian,eTaT H H3onponaHOJi 95:5, OKpacKa: napbi irojia). E>ikjiociiopmh A οτλθληιοτ ot kojiohkh coflepjKa-m«M 2,0% MeTaHOJia ΧΛοροφορΜΟΜ, ijmkjiociiopmh B HfleT npn 2,5% co-flepasaHMH MeTanojia, m UMKJiocnopHH C mojkho ηβητη bo φρβκυ,ΗΜ c 3, 0% MeTaHOJia. ΧροMaTorραφwqecκwe $paKm4H, cosep^anj^e oTflejibHbie KOMFIOHeHTbl UMKJīOCIIOpHKa B qMCTOM BMfle, BbinapHBaiOT B BaKyyMe. TaKHM nyTeM nojiyqafOT 225 mt UMKJiocnopmia A (TO^Ka njiaBJieHHH: 137-i4u°C; l ] D: -189 /MēTaHOJi/; 2o μγ UMKJiocnopHHa B (TO^Ka njiaBJieHHH: 149-151 C) h 52 μγ pMKjiocnopHHa C (ToqKa iuiaBJteHHiī: 150-152°C). ΠΡΚΜΕΡ 2:
OnHcaHHbiM b npHMepe 1 enocodoM npHroTaBJDiBaioT H3 aiiiiapaTa fljiH BCTpHXHBaHHH Kyjn>Typy HHOKyjiaTa. 200 mji θτομ Kyju>Typbi 3a-CeBarjT 5 ūMTpOB nMTaTeJībKOH cpegb! C HKOKVJISTOM 1C, KOTOpbie Ha-X05flTCH B .īīaOOpaTOph'OH yCTaHOBKe JļJIE φβρΜΘΗΤ3Ι1ΙΊΗ ΟΟΒβΜΟΜ 10 jimtpob, h CTepHJiH3yiOT npii 121°C b TeqeHHe 45 MHHyT. Kyjū>Typy nepeMeinMBaiOT npH 25° C c ^hcjiom οδοροτοΒ 750 mn κ npn stom aa-pqpyioT KOJīimeeTBOM B03,nyxa 300 ji/qac.
Ilocjie 72 qacoB $epMeKTau;HM 500 mji nojiyqeHHOH KyjitTyphi HHOKyjiHTa πρημθηημτ ajih saceBaHHH 5 jihtpob mrraTejiBHož cpeflbi FC2, KOTopaa HaxoitHTCs b JiaSopaTopHofi ycTaHOBKe jļhh φβρΜβΗΤ3-φίΜ οοβθμομ 10 ji, η cTepnjiH3yioT πρπ 121 °C b TeqeHHe 45 MMHyr.
CocTaB nirraTejiBHOž cpejļbi c μηοκυομτομ FC2 80 r 40 r 2 r 12 r rjuoK03a ΤρΜΠΚ33ΗΗ
MoqeBMHa cyjii5i|)aT aMMOHMH LV 10500 - /7 - HMTpaT HaTpMH 3 r KMCJIfcJM φΟΟφορΗΟΚΜΟΙΕΗΜ K3JIMM 2 r XJIOpKCTbIM KaJIHH 0,5 r eyj^aT Marnm · 7H^0 0,5 r cyJIbφaτ MeflM ·7Η^0 0,01 r cyjibOaT MapraHpa (II)· 7H^p 0,01 r cyjK^aT «ejiesa (ΙΙ)·7Η^0 0,01 r B 1000 MJI BOflOnpOBOflHO# BOflhl
BejuFiMHjr pH nHTaTejmHoii cpeflbi yeTaHaBJiMBafOT nepefl cTepe-JiMsaiiMeii flo 5,2. IīoceBHyio nepeMeniHBaioT npn 25° C c ^mcjiom o6o-ροτοΒ 750 mmh h npH 3TOM a3pHpyioT KOJureecTBOM B03flyxa 300 jiAiac. CfepMeHTauHio npoflOJLfflaioT b Te^eHHe 144-168 qacoB μ npeK-panpKļT nocjie flocTUKeHHH MaKCMMcuiBHoft KOHpeHTpaiņof ipntiiociiopMHa.
Pfe 4,3 JIMTpa φβρΜΘΗΤαυ,ΜΟΗΚΟΓΟ 6yjl£iOHa, KOTOpbIM κ KOHliy (ļiepMeHTamiii coflepjKHT 500 μγ/mji κομπ^θκοηογο miKJiocnopMHa, bh-flejiHHT pPiKuiiocnopHH A cjieflymmKM oāpa30M.
KneTKH yfia.nHioT m Φθρμθκτ3ιχηοηηογο 6yjibOHa ģMJībTpoBaHHeM, npMJiHnmee Ha KJieTKax coflepaaHHe aHTHOMOTMKa βημηβθμτ 2x1 jiht-poB MeTaHOJia. Pīs προμηβο^ηομ «μακοοτη MeTanoji yflajunoT npn no-HHJKBHHOM flaBJI6KHH Η ΒΚΤΗΒΗΟβ ΒΘΙϋ,ΘΟΤΒΟ 3KCTparnpyiOT H3 ΚΟΗΙίΘΗΤ" paTa 2x250 mji n-6yTMJiaii;eTaTa. Ρί3 φΚΛί>Τρ3Τ3 φβρΜΘΗΤβΙίΗΟΗΗΟΓΟ 0yJibOHa KOMITJieKCHbliii UHKJiocnopHH 3KCTparHpyiOT 2x500 mji n-6yTHJiaueTaTOM. 3KCTpara?bi coesMHHKīT, cyuiaT Ka# 6e3BOflHHM eyjih$aTOM HaTpMS h BbinapHBafOT npH ΠΟΗΜΜΒΗΗΟΜ flaBJieHHH πρκ 40° C. IfoJiyyeHHHŽ Cbipoii npOflyKT (19,2 r) npeflBapMTejibno o^nmajor msto^om χρ0Μ3Τ0Γρ3φκΗ Ha ko-JioHHe Ka 100 r cmuiKarejra 60 (pasMep qacTWLi: 0,063-0,2 mm) c - 18' ~ χ,ποροφορΜΟΜ: MeTaHOJioM: aueTOHOM=92: 4:4 b Ka^ecTBe ajimHTa.
Coaep«amne κομπλθκοηηζ iiHKJiocnopMH φρακι^ΜΗ ( tohkocλοΛημ xpoMaTorpa-$M5ī na-njiacTHHax hs CHJMKarejiH 60 F254, DC-ajifOMMHMe-Baa φοΛΐ>Γ3 /Μβρκ/; KOHCTpyKTūp: reKcaH m aii,eTOH b οτηοιπθημ 2:1, OKpacrca, xjiop m tojimahh) BwnapMBcUor z;ocyxa. Pīs ocTaTKa ot BbinapHBaHHH (5,28 r) OTflejiHfOT μθτοαομ xpoMaTorpa$MH Ha KOJiOHHe HMKJiocnopHH. KojioHHa coflep«HT 115 r cMJiMKareJiH 60 (pasMep ^ac-THiļ: 0,063-0,2 mm,). B Ka^ecTse pacTBopMTejia πρκΜβΗΗΐοτ coiepm-HMe aiieTOHa, noBtimarampe rpasHeHT reKcaHa-aueTOHa. IjHKJiocnopHH A npoMUBaioT coflep»ainHM 23% aiieTOHa n-reKcanoM hs kojiohhbi. Bh-napHBaHHēM coflepjisainnx u;hkjiocnopMH A φρβκ”hh nojiywai0T 1,46 r iiHKjrocnopHHa A. ΠΡΡΓΜΕΡ 3:
Pis Bbipam,eHHOH na cofiepaajueM cojiOflOBUž SKCTpaKT h jiporae-boā SKCTpaKT kocom arape BospacTOM 5-7 βκβΗ KyjiBTypbi niTaMMa Tolypocladium varium sp.no v. 07/93 (NCAIM (P) F 001005) npn-γοτοβληβτ c 5 mji 0,9 %-ηογο pacTBopa xjiopncToro HaTpua cycneH-3HK3 cnop. CycneH3HK3 cnop ΓφΜΜΘΗΗΙΟΤ flJIH ŪOCeBa 800 MJI nHTaTeJIb-hoā cpeflH 1C yKa3aHHoro b npHMepe 1 cocTaBa, KOTopaa Haxo^HTCH b Koaoe 3pjiaHMeMepa οοβθμομ 3 jiHTpa h KOTopan npeflBapKTejiBHO oHJia CTepHjiH30BaHa. Koji6y BCTpHXHBafOT b annapaTe ajih bctphxh-BaHMH (340 mhh , oTKJiOHeKMe 3,5 cm) πρκ 25°C B Te^eHHe 2,5 flHew. IIojiyqeHHOM Kyjn»Typoā HHOKyjiHTa saceBaioT 5 jimtpob nHTa-TejiBHOū cpeflbi FC3. HHTaTejnjHaa cpe^a HaxoflHTCH b JiaāopaTopHo# ycTaHOBKe flJin φβρ.ΜβΗΤ3ΐχΗΗ οοβθμομ 10 ji η Shfjia npeflBapMTejīBHO CTepHJiM30BaKa npM 121°C β τθ^θημθ 45 MHHyT. -/9- LV 10500
CoeTāB nMTaTeJIbHOM cpeflbl C HHOKyJBīTOM 1C copSosa 60 Γ TpHKaSHH . 40 Γ Mo^eBMHa 2 r CyJH^aT aMMOHHH 12 r HM'TpaT HaTpuH 3 r KHCJIbl^ φΟΟφορΗΟΚΗΟΛΜΖ KaJlViPi 2 r ZūOpMCThJpl KaJMft 0,5 r cyjib$aT MarHMfl · 7E£j 0,5 r cy.HbcpaT ieriesa (II)*7HA0 0,01 r B 1000 MJI BOflOlipOBOflHOŪ BOfibi
BejuiqHKy ρΗ πητετθ^κοζ cpeflu TCTaHaBJtuBaior nepefi CTepn-TimaiļViei/i jļo 5,2. noceBHyio KyjibTypy noffinepjKHBaioT τβρΜοοτβτοΜ npn 25° C, ne-peMeiDHBaiūT (1000 mhh ) si a3pnpyiūT 300 jrHTpaMH/^ac B03,qyxa.
KyjibTypy φβρΜΘΗτ^^ίοτ 120-168 ^aeos, nocjie SToro φβρΜβΗ-TaunoHHbrii Ovjiboh no Μκκροδhojiorckhm ηβμθρθηηημ BejranMH coflepūiPīT 600 μγ/μλ κομπλθκοηογο iļHKJiocnopHHa. Pīs oflHoro jnirpa φβ ρ Μβ K T S.IIH Ο Η Η Ο Γ 0 OV'jlbOKa OnMCaHHbiM B ūp!'lM6p6 2 CnOCOuGM nOJiy- naiOT 310 μγ UMKJiocnopHHa A. ΠΡΗΜΕΡ 4:
Jfo BbipaiiļeHKOH na co/tepasameM cojioflOBbiM 3κοτρβκτ n sporae-boh aKCTpaKT kocom arape BospacTOM 5-7 flHeft Kyji.bTypbi niTaMMa Tolypocladium varium sp.no v. cy/93 (NCAIM (P) F 001005) πρκ- -20.- γοτοβληβτ c 5 mji 0,9 %-ηογο pacTBopa xjiopHCToro HaTpMH cyeneH-3hkj cnop. CycneH3MK) cnop πρΗΜΘΗΗκτ flJīH noceBa 800 mji nHTaTejii»-HOM Cpē/ļbi 1C /KaSclHHOFO B npHMepe 1 COCTaBa, KOTOpāfl HaXOČ[MTCH b Kojioe SpJiiiHMeMepa οΟϊθμομ 3 JiMTpa h KOTopan npeflBapMTeJiBHO Obuia CTēpHJiHc-oBaHa. Kojioy BCTpHXMBaioT b annapaTe ajih bctphxm-BaKMH (340 MHH , ΟΤΚΛΟΗΘΗΜβ 3,5 CM) ΙφΗ 25°C B ΤΘΜΘΗΜβ 2,5 flHeft. IIojivvieHHOH KjjībTfījaft HHOKyjiHTa 3aceBaioT δ jihtpob nHTa-TejiBHOž cpeflbi FC4. -īlHTaTejihKaH cpe,qa ηβχοαμτοη b JiaSopaTopKoii ycTaHOBKe rjik φβομθητημμμ oSbcmom 10 λ h SbiJia .npeflBapuTejL&HO cτepwjim3oBaHa πρκ 121 °C b τβ^βΗΗβ 45 MHHyT.
CocTaB nmaTejibHoPi cpeflhi c HHOKyjiHTOM FC4 MajLbTosa 80 r TpnnKasMH 40 r Mo^eBMHa 2 Γ CyJIb$aT aMMOHHH 12 r HHTpaT HaTpMH 3 r KMCJIblM (|)OC(l)OpKOKMCJIbM K3J1MM 2 r XJIOpMCTMŽ H3JIMŽ 0,5 r cyjib$aT MarHHH * 7HJ3 0,5 r cyjū)$aT Mčjļft (11) * 7 Hjp 0,01 r cyju>$aT MapraHita (ΙΙ)*7ΗΛ0 0,01 r cyjib$a-T KcJiē3a (II) -713^0 0,01 r B 1000 MJI BOflOnpOBOflHOM BOflfal īlMTaTeJiBHyK3 cpeay nepes CTepMJiMsaimeM ycTaHaBJiMBaioT
BeJimmbi pH 5,2. IIoceBHyK) KyjiLTypy noflBepraraT $epMeHTaiļim npw 25°C h npM nepeMeniMBaHMH (1000 mmh ) m aapaiiHH 300 JiHTpaMH/Mac - 2/ - LV 10500 b Te^eHMe 144 naeoB. B KOHije φβρΜΘΗΤΕίχΜΜ $epMeHTamioHHbiii čyjii>-oh no ΜΗκροδηοJiorunec kmm μβμθρθηηημ BejmmīH coflepjKMT 628 mt/mji ΚΟΜΠΛβΚΟΗΟΓΟ UHKJIOCnopHHa. Jfe OflHOrO JiMTpa φΘρΜΘΗΤΒΙΙΜΟΗΗΟΓΟ 6yjiBOHa onncaHHbiM b npMMepe 1 ciioco6om nojiyqafOT 305 μγ umkjioc-nopnHa A. ΠΡΜΕΡ 5:
Pi3 BbipaiiieHHon Ha eosepjuameM cojioflOBbiM aKCTpara η κροχχθ-bom SKCTpaKT kocom arape BOspacTOM 5-7 flHež Kyju>Typu uiTaMMa Tolypociadium varium sp.no v. 0Υ/93 (NCAIM (P) F 001005) npn-ΓοτοΒΛΗίοτ c 5 mji 0,9 %-Horo pacTBopa xjiopncToro HaTpus cycneH-3Wo cnop. CycneH3MKD enop πρΜΜθΗΗίοτ sjih noceBa 800 mji miTaTeJib-hoft cpeflbi 10 yKa3aHHoro b npMMepe 1 cocraBa, KOTopan HaxoflHTCH B KOJiče SpjiaHMeftepa οΟβθμομ 3 JiMTpa n KOTopa# npeflBapHīejttHO 6tuia CTepHJiH30BaHa. Koji5y BCTpsxnBaioT b annapaīe ejih Βοτρπχκ-BaHHH (340 MMH , ΟΤΟΟΗΘΗΜβ 3,5 CM) ΠρΗ 25°C Β Τβ^ΘΗΜβ 2,5 AHefi. ĪTojiyqeHHOft Kyjn>TypoM MHOKyjiHTa saceBafOT 5 jihtpob nuTa-TejiBHoit cpeflM FC1. IĪMTaTejiLHaH cpe^a ΗΒχο/ίΗτοΗ b JiaOopaTopHoft yCTaH0BK6 flJIfl φβρΜΟΗΤαΙζΜΗ ΟΟΒΘΜΟΜ 10 JI H ābIJia npeflBapMTēJIBKO CTepnjiHsoBaHa npn 121° C b TeyeHne 45 μηηυτ. IToceBHyio KyjtbTypy noflBepraiOT Φθρμθητβμ,νμ npn 25°C u npn nepeμθiiiMBaHmm (750 mmh ) m aapapMH 300 JiHTpaMH/nae. B 96-m qae £o6aBJiHrOT 2% oīfieJiBHO CTepHJIHBOBaHHOH MaJIBTOSbī, 33ΤΘΜ npOMSBOflHT φβρΜβΗΤβΙίΗΚ) JļO 144~ ro qaca. ®epM6HTaij[iiOHHbjH oyjiBoh' coflepjftnT κ 3T0My MOMeHTy no MMKpo6μojiornneekmm μβμθρθημημ bojimmmh 950 mt/mji KOMriJieKCHoro pnK.7iocnopMHa. Pīs 2,8 JiMTpa φβρΜθKTauMOKHoro 6yjn>0Ka onncaHHbiM b npMMepe 2 cnocočoM BbmejisiOT 2,75 r UMKJiocnopnHa A. LV 10500 1. Cnocoč nOJI^^eHHH KOMITJISKCHOrO aHTHOHOTHKa IļHKJĪOCnopiiHa h/hjih ero ΚΟΜΠΟΗ0ΗΤΟΒ, BKjmmioiiinž K7Jn>THBnpoBaHiie npoflyiļeHTa npn a3po6Hbix ycjioBHHX b πητ3Τθ.ιιβηομ cpe^e, coflepjfiamepi ycBanBa-eMhie hcto^hmkh yrjiepofla, a30Ta m MHHepajīBHbie cojuī c nocjieflyio-npiM BbifleJieHPieM iļejreBbK npoflyKTOB, OTjmarouijiMCH τβΜ, ήό b Ka-qec<rBe npoflyueHTa Hcnojn>3yioT Γρκοοκ Tolypocladium varium NCIM /P/ F 00105, πρκ 3TOM KyjH)THBHpoBaHiie ocymecTBJiHioT πρη ΤβΜΠβ-parype 25-30° 0. 2. Cnocoč no π. 1, oTjm^aramnžcH τθμ, ^to b Ka^ecTBe mcto^-HPiKa B30Ta πρΚΜβΗΗΙΟΤ ΠΘΠΤΟΗ, ΟΤΠΒφβΤ aMMOHMH, TpMiIKa3HH, B Ka-qecTBe HCTOMHHKa yr.nepofla - rjnoK03y, MajibT03y, cop6nT. 3. ΙΙΙγβμπ rpnoKa Tolypocladium varium NCAIM (P) F 001005 - ΠρΟ^ΚΤ ΚΟί/ΟυΐΘΚΟΚΟΓΟ aKTMOHOTHKa īļHKJlOCnOpVind. H/MJĪM 6Γ0 ΚΟΜΠΟ- H6HT0B. LV 10500 ΑΗΗΟΤΑΙίΜΗ Μ3θ6ρβΤβΗΗβ OTHOCHTCH Η Κ Cn0C05y flJIfl nOJiyqeHHH KOMIIJieKC-ΗΟΓΟ UMKJIOCnOpMHa M/'AJIM ero ΚΟΜΠΟΗΘΗΤΟΒ: UHKJIOCnopHHa A, uyiK~ JIOCnopMHa B H UHKJIOCnopMHa C KyJII)THBHpOBaHM0M āMOCMHTe3MpyiOme-ro HassaHHbie 3Ητμ6μοτμκμ nrraMMa rpMČKa b coflepjKaineīi yeBaHBae-Mbie mcto^hmkm yr Jiepo^a m asoTa n MHHepajiLHbie cojīm nMTaTejiBHoft cpeae, npH aapoSHbix ycjiQBwax, μ οτκλοηθημθμ oāpa30BaHHbix npo-flyKTOB. JiJIH ΜΒΟδρβΤΘΗΜΗ 0TJHmMT6J!LbHbIM Πρΐί3Η3Κ0Μ ΗΒΛΗβΤΟΗ TO, qTO npOflyLļMpyK3IIļI'iM ΚΟΜΠΛΘΚΟΗΗΜ 3ΗΤΚ6Μ0ΤΗΚ liMKJIOCnopHH rnTaMM HO-
Boro BHfla rpMāKa Tolypocladium varium, npe^no^TMTēJīBHO αθιίοημ-poBaHHbiH nofl HOMepoM NCAIM(P)F 001005 b BysaneniTCKOM Υημβθρομ-τθτβ caāOBOflCTBa μ πμιπ,θβομ npoMbinuieHHocTH b HaimoHajcbHOM co6-paHHH CeJū)CK0X03fiiiCTBeKHbIX H IipOMbIIlIJieHHbIX MMKpOOpraHHSMOB iiiTaMM Toplypocladium varium sp. nov, 0Υ/93, KyjiLTHBHpyiQT b co-aepjKame# hctomkhkh yrjiepofla, a Taranē opranmecKMe a HeopraHH-yecKne hctovikhkm asoTa u MHHepajīBHbie cojiī! πητβτθοιβηοζ cpese, npH aapoāHbix ycjiOBMsx, npn TeMnepaType 25-30 C, b cjiy^ae Heo6-XOflHMQCTH Bbifle.HHK)T OČ'paSOBaHHblH KOMIIJIcKCKblM aHTHOHOTHK HJIH ΘΓΟ KOΜΠΟK6πTbi M3 φβρΜΒπΤαί^ίΊΟΗΚΟΓΟ 6yJIbOHa Η B Ο.ϋγ^α,θ H600X0flHM0C“ T Κ Ο hM ίΰβΧϊΓ.

Claims (3)

  1. LV 10500 IZGUDROJUMA FORMULA KOMPLEKSĀ ΑΝΤΙΒΙΟΤΙΚΑ - CIKLOSPORĪNA UN/VAI TĀ KOMPONENTU UN SĒNĪŠU KULTŪRAS. TOLTPOCLADIUM YARIUM IEGŪŠANAS PAŅĒMIENS 1. Kompleksā antibiotiķa - ciklosporīna un/vai tā komponentu iegūšanas paņēmiens, kurā ietilpst producenta kultivēšana aerobos apstākļos barotnē, kura satur asimilējamus oglekļa, slāpekļa un minerālsāļu avotus, ar tai sekojošu galaņroduktu izdalīšanu, kas atšķiras ar to, ka kā producentu izmanto sēnīti Tol^pocladium varium N CAIM/P/F 001005, un kultivēšanu realizē 25-3O°0 temperatūrā*
  2. 2· Paņēmiens, saskaņā ar 1. punktu, kas atšķiras ar to, ka kā slāpekļa avotu izmanto peptonu, amonija sulfātu, tripkazīnu, kā oglekļa avotu - glikozi, maltozi, sor-bītu*
  3. 3. Sēnīšu kultūra Tolypocladium varium N CAIM/P/F 001005 - kompleksā antibiotiķa - ciklosporīna un/vai tā komponentu producents.
LVP-93-708A 1988-12-20 1993-06-28 Method for producing of complex antibiotic- cyclosporin and/or it's components and method for producing of tolypocladium varium culture LV10500B (en)

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CA2101777A1 (en) * 1991-01-25 1992-07-26 Toshio Goto Process for producing cyclosporin a and/or c
ATE127528T1 (de) * 1991-04-06 1995-09-15 Dresden Arzneimittel Verfahren zur fermentativen herstellung und isolierung von cyclosporin a sowie neue cyclosporin- bildende stämme.
FI92334C (fi) * 1992-12-30 1994-10-25 Leiras Oy Menetelmä syklosporiinien tuottamiseksi ja menetelmässä käytettävä uusi Tolypocladium-kanta
SI9300303A (en) * 1993-06-08 1994-12-31 Krka Tovarna Zdravil Process for isolation of hypolipemic effective substance
KR100304324B1 (ko) * 1994-10-13 2001-11-22 김용규 사이클로스포린a의제조방법
KR100341355B1 (ko) * 1994-10-14 2002-10-25 주식회사종근당 사이클로스포린a의제조방법
KR0126610B1 (ko) * 1994-10-18 1997-12-29 김충환 고생산 세포융합변이균주에 의한 사이클로스포린 a의 제조방법
EP0725076B1 (en) * 1995-02-01 2001-06-06 National Research Development Corporation of India A process for the preparation of cyclosporin A from tolypocladium species
WO1997001575A1 (fr) * 1995-06-29 1997-01-16 Fujisawa Pharmaceutical Co., Ltd. Substance wf16616, procede de production et utilisation
DE19611094C2 (de) 1996-03-21 1999-06-17 Dresden Arzneimittel Verfahren zur Reinigung von Cyclosporin A und/oder verwandten Cyclosporinen aus einem Cyclosporin-haltigen Rohextrakt unter Anwendung chromatographischer Verfahren mit Kieselgel als Adsorbens
US5747330A (en) * 1996-06-05 1998-05-05 Poli Industria Chimica Antibiotic producing microbe
US5709797A (en) * 1996-06-05 1998-01-20 Poli Industria Chimica S.P.A. Method of isolating cyclosporins
IL143604A0 (en) 1998-12-30 2002-04-21 Dexcel Ltd Dispersable concentrate for the delivery of cyclosporin
US7732404B2 (en) 1999-12-30 2010-06-08 Dexcel Ltd Pro-nanodispersion for the delivery of cyclosporin
US7176001B1 (en) 2000-02-29 2007-02-13 Biocon Limited Manufacture and purification of cyclosporin A
US7696166B2 (en) 2006-03-28 2010-04-13 Albany Molecular Research, Inc. Use of cyclosporin alkyne/alkene analogues for preventing or treating viral-induced disorders
US7696165B2 (en) 2006-03-28 2010-04-13 Albany Molecular Research, Inc. Use of cyclosporin alkyne analogues for preventing or treating viral-induced disorders
EP2151450A1 (de) * 2008-07-29 2010-02-10 Sandoz AG Verfahren zur Aufarbeitung von mikrobiologisch hergestellten zyklischen Oligopeptiden
JP6022548B2 (ja) * 2011-04-28 2016-11-09 セルジーン コーポレイション Ｐｄｅ４阻害剤を自己免疫疾患及び炎症性疾患の治療及び管理に使用する方法及び組成物
GB201319791D0 (en) * 2013-11-08 2013-12-25 Sigmoid Pharma Ltd Formulations

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US4117118A (en) * 1976-04-09 1978-09-26 Sandoz Ltd. Organic compounds
DE2819094A1 (de) * 1977-05-10 1978-11-23 Sandoz Ag Cyclosporin-derivate, ihre verwendung und herstellung
US4215199A (en) * 1978-06-05 1980-07-29 Sandoz Ltd. Antibiotic production
SE448386B (sv) * 1978-10-18 1987-02-16 Sandoz Ag Nya cyklosporinderivat, forfarande for framstellning av dem samt farmaceutisk komposition innehallande dem
US4764503A (en) * 1986-11-19 1988-08-16 Sandoz Ltd. Novel cyclosporins

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NO175873C (no) 1994-12-21
DK640989A (da) 1990-06-21
EP0379708B1 (de) 1994-12-07
RU1836425C (ru) 1993-08-23
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GB2227489A (en) 1990-08-01
PL162959B1 (en) 1994-01-31
DD290356A5 (de) 1991-05-29
SE8904262D0 (sv) 1989-12-19
GB8928715D0 (en) 1990-02-28
YU47100B (sh) 1994-12-28
GB2227489B (en) 1992-09-16
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FR2640641A1 (fr) 1990-06-22
DE58908740D1 (de) 1995-01-19
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NO175873B (lv) 1994-09-12
UA13320A (uk) 1997-02-28
FI92603C (fi) 1994-12-12
SE505442C2 (sv) 1997-08-25
CA2006144C (en) 1995-02-21
NO895160L (no) 1990-06-21
US5156960A (en) 1992-10-20
FR2640641B1 (fr) 1991-05-10
FI896157A0 (fi) 1989-12-20
CA2006144A1 (en) 1990-06-20
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BG60274B2 (en) 1994-03-31
YU241589A (en) 1991-02-28
IT8922765A0 (it) 1989-12-20
LV10102B (en) 1995-02-20
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JPH0763356B2 (ja) 1995-07-12
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