CS254131B1 - Method of carnitine dehydrogenase production - Google Patents
Method of carnitine dehydrogenase production Download PDFInfo
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- CS254131B1 CS254131B1 CS225284A CS225284A CS254131B1 CS 254131 B1 CS254131 B1 CS 254131B1 CS 225284 A CS225284 A CS 225284A CS 225284 A CS225284 A CS 225284A CS 254131 B1 CS254131 B1 CS 254131B1
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- nph
- efl
- carnitine dehydrogenase
- hoctb
- pseudomonas
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- 238000000034 method Methods 0.000 title claims 3
- 108010029922 carnitine dehydrogenase Proteins 0.000 title claims 2
- 238000004519 manufacturing process Methods 0.000 title claims 2
- 241000589776 Pseudomonas putida Species 0.000 claims description 4
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 claims 1
- 238000012258 culturing Methods 0.000 claims 1
- 239000007788 liquid Substances 0.000 claims 1
- 229910052760 oxygen Inorganic materials 0.000 claims 1
- 239000001301 oxygen Substances 0.000 claims 1
- 241000589516 Pseudomonas Species 0.000 description 5
- 101150096418 Mepe gene Proteins 0.000 description 3
- 101100321306 Bacillus subtilis (strain 168) yxeB gene Proteins 0.000 description 1
- 235000006693 Cassia laevigata Nutrition 0.000 description 1
- 241001553014 Myrsine salicina Species 0.000 description 1
- 229910017917 NH4 Cl Inorganic materials 0.000 description 1
- 229930182556 Polyacetal Natural products 0.000 description 1
- 241000522641 Senna Species 0.000 description 1
- 240000003834 Triticum spelta Species 0.000 description 1
- 239000012491 analyte Substances 0.000 description 1
- 206010009887 colitis Diseases 0.000 description 1
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 description 1
- 229910000396 dipotassium phosphate Inorganic materials 0.000 description 1
- 235000019797 dipotassium phosphate Nutrition 0.000 description 1
- 230000000422 nocturnal effect Effects 0.000 description 1
- 125000001400 nonyl group Chemical group [H]C([*])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])[H] 0.000 description 1
- 229920006324 polyoxymethylene Polymers 0.000 description 1
- 229940124513 senna glycoside Drugs 0.000 description 1
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- Enzymes And Modification Thereof (AREA)
- Organic Low-Molecular-Weight Compounds And Preparation Thereof (AREA)
Description
i
HA3BAHHE H30EPETEHHH Μθτοα nonyneHHH KapHHTHHflerHflporeHaabi (ΚΦ 1.1.1 *108)
06nacTh npHMeHeHHH H3O6peTeHHH Η3θ6ρβΤβΗΗβ OTHOCHTCH K MeTOfly nOJiyneHHH KaHHTHHflerHflpOreHa3bl, KOTOpaH MO“Μβτ npHMeHHTbCH B KflHHHKO-XHMHWeCKOift flHarHOCTHKe «JIH OnpefleneHHH L-KapHH-THH3.
XapaKTepHCTHKa H3BecTHbix TexHHHecKHx pemeHHň' flo HacToamero ΒρβΜβΗΗ KapHHTHHflerHAporeHaaa He npoHSBOflHTCH.B xayecTBe kom-MepuecKoro npenapaTa. HccneflOBaHHH b HanpaBneHHH npHMeHeHHH «jih κοηκηβοτΒβΗ-Horo OnpefleneHHH KapHHTHHa oTcyrcTByioT. B HacTonmee BpeMH L-xapHHTHH kojih-HeCTBeHHOiOnpeflejIHIOT 4epMeHTaTHBHbIM Cnoeo6OM C nOMOmblO KapHHTHH-aueTHJITpaH-c<t>epá3bi (ΚΦ 2.3.1.7)) b κομ6ηηβαηη c aflHn-CoA-cHHTeTa3oň (ΚΦ 6.2.1.2.) hjihflTHB (Pearson, D.J., Tubbs, P.K., Chase J.E.A. b Methoden der tnzymatischenAnalyse (pefl. H.U. Bergmeyer), Akademie-Verlag Berlin 1970, c. 1710-1723).06a MeTOfla o6naflaioT pnflOM HeflocTaTKOB, H3 κοτορωχ, πομημο 3HanHTenbHbix 3βτ-par, cneflyeT ynoMHHyTh cpaBHHTenbHO cnoxHyro MeTOflHKy h BbicoKyio BOcnpm&niH-BocTb no OTHOmeHHio κ MeniaiotuHM bjihhhhhm. ílocne Toro κβκ 6blná npeflocTaBJíeHaBO3MOxttocTb KonnuecTBeHHoro onpefleneHHH L-KapHHTHHa c noMOiftbio onTHaecKoroaHanH3a (Sčhopp, W., Analyt. Biochem., nyĎnHKaflHH noflroTaBnH^aeTcn)γ cnefly-βτ paccHHTbfflaTb Ha mnpoKoe npHMeHeHHe KapHHTHHflerHflporeHaabi. ΦβρΜβΗτ oĎpaay-eTCH b pa3*nHHHbK BHflax Pseudomonas (Aurách., H., Kleber, H.-P., Schopp, W.,Biochem. Biophys.Acta, 139, 505 (1967); Kleber, H.-P.j Seim, H., Aurich, H.,Straek, E., Arch. Microbiol. 116, 213 (1978)). 06HapyxeHHbie yflenbHbie ηκτββ—HOCTH φβρΜβΗΤΒ B ÓeCKneTOHHblX 3KCTpaKTaX ΗΜβΙΟΤ 3HaneHHH MeJKfly 1 H l,5efl./Mr.
OrmcaHHe nareHTa C1HA 4 221 869 onncbiBaeT Μβτο« nonyneHHH xapHHTHHflerHflpore-Haabi noepeflCTBOM KynbTHBHpoBaHHH mTaMMa Pseudomonas C1P52 191 b οδωηΗωχycnoBHHX KyňbTHBaflHH πρκ 30°C 6oneé 24 qacoB. EtonynaeTCH φβρΜβκτ c βκτηβ—HOCTb» 0,55 efl./ΜΓ., JtenneTCH oneBHflHMM, hto npoflyKT co cronb HHsxoň βκτηβ— 254131
HocTbio He npeflCTaBjíHeT HHTepeca b KOMMepnecKOM οτΗοιηεηΗΗ. 3φφεκτΗΒΗΜ& Μβτορohhcťkh. φβρΜβΗτβ b pHTHpyeMOM πητθητηομ onHcaHHH He yKaabiBaeTca.
Uejib H3o6peTeHHfl ΗΗ3ο6ρβτβΗΗβ ΗΜββτ CBoeň uejibio npeflCTaBHTb MeToa nonyneHHH KapHHTHHflerHflpo-reHa3bi, b xoae κοτοροτο noc/ie ae3HHTerpanHH κηβτοκ npocTbiM cnocoSoM yxeB 3KCTpaKTe SejiKOB nocTHranacb 6bi aKraSHocn φβρΜβΗτβ no MeHbmeň Mepe b3 efl./ΜΓ H 6bUIO 6bl BO3MOXHO 3KOHOMHHHO OCytUeCTBHTb βΓΟ OUHCŤKy BOJlOTb RO3Ha'<ieHnň 25 efl./ΜΓ c noMombio HeMHoronHcneHHbix, oflHaxo xe o6naflaiomHX 3φφβκ-THBHblM fleňCTBHeM ΜΘΤΟΑΟΒ pa3fleJieHHH, Πομημο 3τογο, nonxeH 6wrb nojiyqen npenapaT, Ko-ropbiň npaKTmecKH He coflepXHThh MenaTflerHflporeHa3bi, hh naKTaTperHflporeHaabi.' 3το ΗΜββτ cymecTBeHHoe 3Ha-ηβΗΗβ, Tax K3K τβΜ caMbiM b nonHoň Μβρβ npoHBnnioTCH 3HanHTejibHbie npeHMymecT-Ba MeTOfla onpeneneHHH L-KapHHTHHa c noMombio KapHHTHimerHflporeHa3bi πρκ homoiuhonTHHecKoro aHanH3a, npeftnoxeHHoro W.SchÓpp. H3JIOXeHH6 CymHOCTH Η3θ6ρβΤβΗΗΗ Η3θ6ρβτέΗΗβ ΗΜββτ CBoeň 3ananeň nocpeflCTBOM Bbióopa MHKpo6HanbHoro HCTOHHHKa*φβρΜβΗΤ3, a TaKXe nOCpeflCTBOM ΠρΗΜΘΗβΗΗΗ HOBblX yCJlOBHň KyjlbTHBHpOBaHHH BOB3aHMOCBH3H C 3φφβΚΤΗΒΗθή ΤβΧΗΟΠΟΓΗβή OqHCTKH npeflOCTaBHTb B ρββΠΟρΗΧβΗΗβxejiaeMbiň φβρΜβΗτ c Bbicoxoň ynenbHoň aKTHBHocTbio, He coflepxamnň MemaioiUHX no-CTOPOHHHX φβρΜΘΗΤβΤΗΒΗΜΧ 3KTHBH0CTeň.
CornacHo Η3θ6ρβτβΗΗΐο, 3Ta 3aflaqa pemaeTCH τθμ, ητο φβρΜβΗτ KapHHTHHflerHgpo-reHa3a nonynaeTCH nocpeflCTBOM KynbTHBHpoBaHHH nrraMMa Pseudomonas putidaATCC 17633. 3τοτ ιητβμμ, κοτορωή xpaHHTcn Taxxe h b UeHTpanbHOM HHGTHTyTeΜΗκροδΗοηοΓΗΗ h 3KcnepHMeHTanbHoň TepanHH b ΝβΗβ h nonýHHJi TaM o6o3HaqeHHezIMET 10947, cornacno Η3θ6ρβτβΕΗΚ>, KynbTHBHpyeTCH πρκ orpaHHneHHoň nopaneKHcnopofla. B neTanbHOM HŠnoxeHHH: πρβΑβτβΒηβΗΟ ΒωποηΗβΗΗβ MeTOfla nocTa-flHHHO.
ConepxaHHe nrraMMa npoH3BQflHTCH npeHMymecTBeHHO Ha 2%-hom. arape πρκ 4°C bτβΜΗοτβ h npH Hcnonb3OBaHHH bxanecTBe HCTOHHHKa yrnepofla-flekaHa h b xanecTBeHCTOHHHxa a3OTa - (NH^JjSO^. KynbTHBHpoBaHHe Pseudomonas putida Ha KapHHTHHenpoH3BOflHTCH nyTeM nepeceea ot KynbTypbi-npeflmecTBeHHHKa, KOTOpaa 6bina nony-ueHa, HanpHMep, πρπ πρΗΜβΗβΗΗΗ ra3Oo6pa3Horo reKcaHa. KynbTHBHpoBaHHe npoH3-βοαητοη πρκ 20-40°C b norpyxeHHoň xynbType b xHflKoň MHHHManbHoň cpene. Bm-paniHBaHHe, cornacHO Η3θ6ρβτβΗΗΐο, npoBOflHTCH b tskhx ycnoBHnx aspaqHHj ko-Topbie COOTBeTCTByiOT CHHbHO OrpaHHUeHHOMy CHaÓxeHHK) ΜΗΚΡΟΟΡΓ3ΗΗ3ΜΟΒ KHCJIOPO-flOM. ΠρΗ 3THX yCJIOBHBX ΤβΚΧβ H aKTHBHOCTb KapHHTHHaerHflP0reHa3b! flOCTHraeTMaKCHMajihHbix 3HaneHHň. B KaHecTBe HCTOHHHKa a3OTa πρΗΜβΗΗΐοτ (NH4)?S04, NH^Cl η τ.π. nočně BpeMeHH KyjibTHBayHH, cocTaB/unomero, b 33bhchmocth ot φπ-3HOJiorHuecKoro cocTOHHHH h KOJiHuecTBa noceBHoro MaTepHaua, 5-40 nac . , 6axTe-PHanbHym Maccy BbmensioT H3 KynbTypanbHoň epeflbí (HanpHMep, πρκ πομοπιη ηθητρη-φyΓHpOBaHHH) Η B 33ΚηΚ>ΗβΗΗΘ, Ηβ ΠρΗ6βΓ3Η K npOMexyTOMHOH ΠρΟΜΗΒΚβ, pecyCneH“flHpywT β φοεφβτΗΟΜ δνφβρβ. nonyneHHe HeonHmeHHoro δθεκηβτοηΗΟΓΟ SKCTpaKTa npoH3BOflHTCH npeHMymecTBeHHOnyTeM 3aMopaxHBaHHH c nocnenyiomHM οττ3ηβθηηθμ, a 3βτβΜ ^eHτpHφyΓHpoBaHHH.BbicoKas 3φφβκτΗΒΗοβτΒ 3τογο npocToro MeTOfla ae3HHTerpaiíHH κηβτοκ μοχθτ 6biTbocoBeHHo xopomo npopeMOHCTpHpoaaHa npH cpaBHeHHH c οδρηδοτκοή yjibTpa3ByxoM(TaSnHua 1) . 254131
CoflepxamaHCH b 6εεκηβτοΗΗθΜ sxcTpaxTe 6ε.πκοΒ KapHHTHHAerHAporeHaaa MoaceT6biTb ošuměna b 3-5 pa3 nocpe^CTBOM ýpaKUHOHHoro BbicanHBaHHH h, εοοτΒβτοτ-ΒβΗΗΟ, 3KCTpaKIJHH, HanpHMep, npH ΠΟΜΟΠΙΗ CyjlbiaTa aMMOHHH. fljIH 3ΤΟΓΟ κ 6en-κοβομυ 3KCTpaKTy Ao6aBJiHioT TBepflhift cyjibifcaT ημμοηηη no κοηθίηοΑ KOHneHTpa-Hhh, cooTBeTCTByromeň 70% HacbímeHHH, npHneM bch aKTHBHocTb nenHKOM OKa3biBaeT-ch b oca^Ke. Hnn flanbHeňmeň omhctkh KapHHTHHflerHflporeHaabi octstok nepeMemn-BaioT c pacTBopoM cyabiJiaTa ^μμοημη, coflepwanwM cooTBeTCTBeHHO 0,05 Mojib/nNa2HPO4, npn 0°C c ποηημθηηθμ KOHueHTpauHH (NH4)2S04, Kaawbiň pa3 cooTBercT-ΒβΗΗΟ no 5 MHHyT. ΦβρΜβΗτ πρπ 9TOM Bo Bce Sojibuieň Mepe noABepraeTCH paCTBO-peHHio. OcHOBHoe KOJiHuecTBo pacTBopaeTCH b flHanaaoHe ROHueHTpaiwň cyjib<t>aTaβμμοηηη, cocTaBJíHiomHX MejKfly 50 η 45% HacbnneHHH; KapHHTHHflerHflporeHa3a mo-χετ nenocpeflCTBeHHO H3 3thx pacTBOpos 6biTb OHHineHa flo yjjeJibHOfí hkthbhocth,COCTaBJlHromeň npH6jIH3HTeJlbHO 25 βΑ./μγ ,nOCpeflCTBOM ΠρΗΜβΗβΗΗΗ ΓΗΛροφοβΗΟήxpoMaTorpaij)HH, HanpHMep, Ha 10-Kap6oxcHfleuHJi-Ce4»apo3e.
TexHHKO-sKOHOMHHecxHe npeHMymecTBa npeflJio«eHHoro npoyecca coctoht b tom,mto uiTaMM Pseudomonás putída ATCC 17633 yace hcxoaho no cpaBHeHHio c H3BecT-HbIMH WTaMMaMH npOHSBOflHT B HeCKOJIbKO pa3 60JlbmyiO (JiepMeHTaTHBHylO aKTHB“ ·HocTb. 3to cneflyeT othgcth KaK.Ha cneT cBoeo6pa3HH npHMSHeHHoro niTaMMa, ko-Topbiň OTJiHHaeTCH sthm ot Bcex flpyrnx HCCJieAOBaHHbix ιπτ3μμοβ Pseudomonás Pe-tida, Tax h Ha cm oco6mx yenoBHň aapaijHH, to ecTb orpaHHueHHoro CHaSxeHHHKHCJiopoflOM. flanbHeňniHe npeHMymecTBa jjaioT μθτοα AeaHHTerpanHH κπβτοκ npH no-ΜΟΠΙΗ 3aM0pa»HBaHHH H OMHCTKa φβρΜβΗΤβ Ha 1Ο-ΚβρδΟΚΟΗΑβμΗη-ΟβφβρΟΒε.nonyHeHHbiň b cooTBeTCTBHH c Η3θ6ρετβΗΗβΜ φερΜβΗτ oSnanaeT xopomeň cToňKoc-Tbio npH xpaHeHHH. ΠρΗ yflejibHoň axTHBHOCTH 10-20 θα./μγ MoryT o6HapywHBaTbCHMaKCHMajIbHO CJieflyiOfflHe nOCTOpOHHHe φβρΜβΗΤ3ΤΗΒΗΜΘ axTHBHOCTH:
ManaTflerHflporeHa3HaH h JiaxTaTflerHflporeHa3HaH: xajKflaH MeHee 0,1%. B Hracecjie-flyiomeM H3JioxeHHH Η3θ6ρετβΗΗβ aojekho 6ητβ 6onee AeTanbHo οΘέηοηθηο Ha oahomnpHMepé ocymecTBneHHH.
npHMep ocyigecTBneHHH .KyjibTHBHpoBaHHe Pseudomonás putída ATCC 17633 ócymecTBjíHeTCH b cpefle, koto-;paa b 1 jiHTpe coflepacHT cneflywnme HHrpeflHeHTbi: K2HPO4 - 7,46 r; NaH2P04 - 0,54rr; MgS04-7H20 - 20 μγ; CaCl2-2H20 - 20 μγ;Fe(NH4)2(S04)2-6H20 - 10 μγ; b xanecTBe HCTOHHHxa a3OTa cnyxHT (NH4)2S04 (1 r/π),b xanecTBe HCTOMHHKa yrnepofla - D, t-xapHHTHH , HC1 (5 r/n). . BaxTepHH npeflBapHTenbHo KynbTHBHpyroT Ha τοή xe cpe«e c Hcnojib3OBaHHeM b xa-uecTBe HCTOHHHxa yrnepoAa-ra3Oo6pa3Horo rexcaHa (0,015% b noAaeaeMOM BO3fly-xe). IlěpeceB npoH3BOAHTCH b TeneHHe ηοΓβρΗφΜΗΜβοκοή φβ3ΐ>ι pocTa, onTHMeckaHnnoTHQCTb npceBHoro MaTepnana cocTaBJíaeT npH6nH3HTenbHo 0,1 no .KyjibTHBHpoBaHHe npOBOflHT B 1 0-JlHTpOBOM φβρΜΘΗΤβρβ C OSbeMOM 3arpy3KH7 JIHTpOB npH HHTeHCHBHOM ΠβρβΜβΠΙΗΒΗΗΗΗ H nOflaue BOSflyxa, COCTaBJIHK>meH 2-10 Ji/nac, Πό HCTeneHHH BpeMeHH KynbTHBHpoBaHHH, cocTaBJíHioinero 20-30 nacoBnpH 30°C, Korná noMyTHeHHe xynbTypbi cocTaBJíneT E^ »1,5-2,5, 6aKTepHanbHyioMaecy ΟΤΑβΠΗΙΟΤ ΑβΗΤρΗφνΓΗρΟΒΗΗΗβΜ Η B 3βΚΠίΟΗβΗΗβ peCyCneHflHpyiOT B 0,05MOJIb/nφοοφετΗΟΜ δνφερβ pH 8 (E npH6jiH3HTenbHo paBHa 20) . 100-mji nopHHH 3Toň cycneH3HH 3aMopaxHBaiOT npH -80°C H BbiAepxHBaioT πρπ 3ToňŤeMnepaType b TeweHHe 60 MHHyT. 3aTeM npoóbi BbiAepxHBaioT b TeneHHe 4 nacoBnpH 30eC. Ilocne AeHTp^yrHpoBaHHH πρκ lOOOOxg b τβΜβΗΗβ 30 MHHyT no MeHbmeňMepe 75% 3kthbhocth jcapHHTHHflerHflporeHa3bi ηβχοαητοη b cynepHaTatcce. «a cn- 254131 flepwameM κπβτοκ. YflenbHaH aKTHBHOCTb φβρΜβΗτβ b sthx ycnoBHHx HMeeT 3Hane-hhh 2-4..en./μγ, coflepacaHHe 6entca cocTaBJíneT πρκ6πΗ3ΗτεπΒΗθ 5 μγ/μπ. H3 βτογο pacTBopa ocHOBHoe KOjnmecTBÓ -6enKa ocax«aioT npn 4°C nocpeflCTBOM ho-SaBJíeHHH cyjibifraTa ημμοηηη no κοηθμηοη KOHijeHTpauHH, cooTBeTCTByiomeň 70%HaebímeHHH. Bca aKTHBHOCTb KapHHTHHflerHflporeHa3bI HaXOflHTCH B ocaflKe. IlocneijeHTpHttiyrHpoBaHHa ocanox nofleepraioT pacTBopeHKo b pacTBOpax cynbiJiaTa aMMO-HHH (c yMeHbmaioiueňCH ero KOHtieHTpaijHeň b 0,05 Monb/n Na^HPO^.) npH 0eC c ποβ-tophkjhhmch pecycneHnHpoBaHHeM h ijeHTpn<})yrnpOBaHneM. TaxnM nyTeM 90% eceňaKTHBHocrn KapHHTHHflernflporeHa3bi 0Ka3biBaeTca b cynepHaTaHTeflHana3OHa κοΗΐχβΗ—TpaijHň Me««y 50 h 45% HaebímeHHH (cm. TaSnHqy 2). Φθρμθητ (ynenbHOň aKTHBHOC—th 5-10 efl./Mr) mojkho CHOBa ocaflHTb flo6aBJieHHeM cyjib<i>aTa ημμοηηη flo κοΗβη—hoh KOHixeHTpauHH, cooTBeTCTBywmeň 70% HaebímeHHH. Ecjih nonyneHHMň SKCTpaKUueňb cpeae cyjib4>aTa aMMOHHH cynepHaTaHT (C 45) ranecTH Ha KonoHKy, HanonHeHHyio10-Kap6oKCHaeitfui-Ce<i>apo3oň, h anroHpoBaTb pacTBopaMH cyjibifcaTa ημμοηηη (40, 35 h 30% HaebímeHHH) npw 22°C, to aocTHraeTCH onHCTKa KapHHTHHnerHnporeHasbipo yflejibHoň βκτμβηοοτη, cocTaBJíHmmeň πρη6λη3Ητθπβηο 25 θλ./μγ (Ta6mma 3).
TaSjíHua 1 nojiyneHHe KapHHTHHflerHnporeHa3bi H3 Ps. putida npH Hcnonb3OBaHHH paanHHHbixΜβτοβΟΒ «e3HHTerpaijHH κπβτοκ
BaxTepHanbHyio Maccy nocne KynbTHBHpoBaHHH na D, L-KáPHHTHHe οτηβηπιοτ npwnoMomn ueHTpHifcyrHpOBaHHH, pecycneHflHpyioT b 0,05 Monb/π φοοφβτΗΟΜ 6yφepePH 8 (e££ iiph6ji. 20) h aaTeM no 20 μπ cycneH3HH κπβτοκ nonBepraioT ne3HH-
TerpayHH - o6pa6oTKoň yjibTpa3ByxoM (4 MHHyTbi npH 50 Bt/cm1) ηπη xesaMopá- χηββηηθμ (30 MHHyT npH -80°C) n pa3M0paxHBaHHeM (1 nac npn 30°C) Μβτοχξ pe3- 06maa 06ma« aK- 06qee yflenbHan HňTerpaijHH aKTHB- THBHOCTb κοπή— - aKTHBHOCTb κπβτοκ HOCTb b cynepHa- necTBO b cynepHa- nocne TaHTe nocne Senná b TaHTe fleŠHH- fleaHHTerpa- cynepHa- (efl./Mr) TerpaqHH HHH Η Β(βΗΤ- TaHTe κπβτοκ ΡΗφγΓΗρΟΒβ- (μγ) (en.) HHH npH15000xg (efl.)
YnbTpa3ByK 180 177 120 1,5 3aMopaxHBaHHe/ pa3MOpaXHB3HHe 177 183 80 2,3 254131
TaOjiHua 2 Φρβκμ»θΗΗ3Α sKCTpaKHKH KapHHTHHflernflporeHa3M b cpefle cynb$aTa βμμοηηη flaHHbte othochTch k 320 μπ Βκοτρβκτβ Οβπκοβ (4,5 μγ 6enxa b 1 μπ), κοτορωή6bin nonytieH nyTeM aaMopaxHBaHHn/pasMOpaxHBaHHn. (NH4)2S04(% HacbímeHHH) OSteM (μπ) Akthb- HOCTb (éfl.) Ββποκ (μγ) ynenbHaH aKTHB- HOCTb (efl./Mr) Bbixofl (X) 60 (ocaflox) 3950 1700' 2,3 60 (cynepHataHT) 50 55 (C 1) 57 35 0,8 55 (C 2) 50 105 2,6 55 (C 3) 50 40 120 0,3 1,0 45 (C 1) 54 1144 178 6,4 29 45 (C 2) 52 1548 210 7,4 39 45 (C 3) 42 670 110 6,1 17 35 (C 1) 48 220 585 0,4 5,5
Ta6nnqa 3 O^HCTKa KapHHTHHflerHflporeHaaw Ha IO“Kap6oKCjífleuHJi~Ce4iapo3e
Ha KonoHKy, κοτορβΗ coflepxHT 18 μπ 10-Kap6ÓKCHfleii;HJi-Ce4apo3bi, ypaBHOBeineH'HOH paČTBOpOM ΟγΠΒφβΤΑ aMMOHHJT (45% HaCbímeHHH) , HaHOCHT 106 ΜΠ paCTBOpaKapHHTHHflerHflporeHa3bi 8 pacTBope cynM>aTa πμμοηηη 45%-ηογο HacbnneHHH, Pac-TBop coflepxHT 2080 eflHHHfl aKTHBHOCTH φερΜβΜτπ h 329 μγ 6enKa. 3πιοηηη npo-H3BOflHTCH ΠΡΗ 22°C,.
OpakflHH(» n/n) ΜΠ (NH4),S04(% Hacbl-CbímeHHH) Akthb- HOCTb (efl.) Ββποκ (μγ) yflenbHan aKTHB“ HOCTb(efl./Mr) . BbIXOfl (Z) 1 í-- 1 3 4 s 7 1 106 45 22 2 25 45 2,2 254131 1 ripoflonaceHHe Ta6n. 3 1 2 3 4 S é ' 7 3 ' 25 45 · 0,6 4 25 40 1,3 5 25 40 1,9 6 25 35 43 7 25 35 177 25- 7 8,5 8 25 35 225 18 12,5 11 9 25 35 167 17,5 8,5 8 10 25 30 386 19 20 18,5 11 20 30 546 22 25 26 12 25 30 520 46 II 25
ΦορΜγΠβ H3OfipeT€HHB 1. MaTOfl Η3ΓΟΤΟΒΠβΗΗΗ KapHHTHH«erHflpOreHa3M nyTeM KyjIbTHBHpOBaHHH Pséúdo-monas putida b norpyxeHHoft xynbType b xHflxoň MHHHManbHofl nHTaTenbHOň cpe-«e, oTJiHQaioiunňcH τβΜ, hto ιπτβμμ ATCC 17633 BbipamireaioT np« orpaHnueHHOMCHaBxeHHH KHCJIOpOflOM. 2. ΜβτοΛ, cornacHo nyHXTy 1., OTnHHaiomHňca τβΜ, hto nocTynneHHe Bosayxa coc~τβΒΠΗβτ ot 0,3 no 1,5 mrrpa b uac Ha jihtp πΗτβτβηΒΗοή cpeaw. 3. Me-roa, cornacHo nyHKTaM 1. h 2,, OTJinuaiomHňcH τβΜ, hto fleanHTerpanHH tκπβτοκ ocymecTBJíaeTca npH πομοικη 3aMopaxHBaHfta/pa3MopaxHBaHHa. AHHOTaHHg
HaoČpeTetraexo«e xoTo-GenxoB noc- HCTOHHHka Η3ο6ρβτβΗΗβ othochtch κ MeTpny nonýMeHHfl KapHHTHHflerHflporeHa3M. ΗΜββτ uejibto npefljioxHTb MeTOfl nonyneHMa kapimTWiaerHAporéRasN, bporo nócne «e3HHTerpaitf«i χπβτοχ npocTUM ΜβτοαοΜ yxe b.axcTpaxTeTHrajiacb 6w ηκτηβηοοτβ φβρΜβκτβ no MeHbmeň Mepe b 3 en./Mr, 3aflaneň Η3θ6ρβΤβΗΗΗ HBJIHeTCH, nOCpeflCTBOM ΠβΗΜβΗβΗΗΗ ΜΚΚροβΗΟΓΟ φβρΜβΗΤΗ COBM6CTHO C 3φφβΚΤΗΒΗθή ΤβΧΗΟΠΟΓΗβή OHHCTXH, CflejiaTb aOCTynHMM~ ’,xenaeMbiň φβρΜβΗτ c ewcoKoň ynejibnoň axTHBHocTbio. 3ra sanana pemaeTca τβΜ,hto φβρΜβΗτ xapHHTHHnerHflporeHasy ΒΗΛβΛΗΜτ H3 niTaMMa Pseudomonás putidaATCČ 17633.
IlpH3HaHo Η3θ6ρετβΗΗβ no pe3ynbTaTaM 3xcnepTH3hi, ocymecTBneHHoň Beflow-CTBOM no nenaM Η3θ6ρβΤβΗΗή Η ΠβΤβΗΤΟΒ ΓβΡ. 254131 1
Claims (4)
1. Způsob výroby karnitindehydrogenázy kultivací Pseudo-monas putida v submersní kultuře v tekutém minimálnímživném médiu, vyznačující ee tím, že se kmen ATCC 17633kultivuje za omezeného přívodu kyslíku·
2. Způsob podle bodu 1, vyznačujícívzduchu je v množství od 0,3 do 1,5Živného prostředí. se tím, že přívod1 za hodinu na 1.1
3. Způsob podle bodu 1, vyznačující se tím, že dezintegracebuněk se uskutečňuje zmrazením a rozmražením. uieorodaky výrobnž-polygrafický podnik,' Proektnaja
4, UŽgorod»3896 , Cena 2,40 Kcs
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| DD24909083A DD228974A3 (de) | 1983-03-23 | 1983-03-23 | Verfahren zur herstellung von carnitindehydrogenase |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CS254131B1 true CS254131B1 (en) | 1988-01-15 |
Family
ID=5545844
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CS225284A CS254131B1 (en) | 1983-03-23 | 1984-03-28 | Method of carnitine dehydrogenase production |
Country Status (3)
| Country | Link |
|---|---|
| BG (1) | BG49226A1 (cs) |
| CS (1) | CS254131B1 (cs) |
| DD (1) | DD228974A3 (cs) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP3059735B2 (ja) * | 1989-10-13 | 2000-07-04 | 旭化成工業株式会社 | Ｌ―カルニチンデヒドロゲナーゼおよびその製造法 |
-
1983
- 1983-03-23 DD DD24909083A patent/DD228974A3/de not_active IP Right Cessation
-
1984
- 1984-02-13 BG BG6423684A patent/BG49226A1/xx unknown
- 1984-03-28 CS CS225284A patent/CS254131B1/cs unknown
Also Published As
| Publication number | Publication date |
|---|---|
| DD228974A3 (de) | 1985-10-23 |
| BG49226A1 (en) | 1991-09-16 |
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