(+-)-Methionine → Penicillin G Potassium

HRID 716431

Reaktionsgleichung

Reaktionsgleichung von HRID 716431
O=[CH:2][C@@H:3]([C@H:5]([C@@H:7]([C@@H:9]([CH2:11][OH:12])O)O)O)O.O.O.O.O.O.O.O.S([O-])([O-])(=O)=O.[Mg+2].S([O-])([O-])(=O)=O.[NH4+].[NH4+].P([O-])(O)(O)=O.[K+:38].[CH3:39][C:40]1[N+](CC2C=NC(C)=NC=2N)=CSC=1CCO.Cl.[Cl-].OC(CCC[CH2:65][C@H:66]1[C@@H:74]2[C@@H:69]([NH:70][C:71]([NH:73]2)=[O:72])[CH2:68][S:67]1)=O.NC([C:81]([OH:83])=[O:82])CCSC.[C:84](=O)([O-])[O-].[Ca+2].C1[C@H](N)[C@@H](O[C@H]2O[C@H](CN)[C@@H](O)[C@H](O)[C@H]2O)[C@H](O)[C@@H](O[C@H]2O[C@H](CO)[C@@H](O)[C@H](N)[C@H]2O)[C@@H]1N.CCCCCCCCCCCCOS([O-])(=O)=O.[Na+].CCN(C1C=CC(C(C2C=CC(NC3C=CC(OCC)=CC=3)=CC=2)=C2C=CC(=[N+](CC3C=CC=C(S([O-])(=O)=O)C=3)CC)C=C2)=CC=1)CC1C=CC=C(S([O-])(=O)=O)C=1.[Na+]>O.O.O.O.O.O.O.S([O-])([O-])(=O)=O.[Fe+2].O.O.O.O.O.S([O-])([O-])(=O)=O.[Mn+2].O>[CH3:84][C:66]1([CH3:65])[S:67][C@@H:68]2[C@H:69]([NH:70][C:11]([CH2:9][C:7]3[CH:5]=[CH:3][CH:2]=[CH:39][CH:40]=3)=[O:12])[C:71](=[O:72])[N:73]2[C@H:74]1[C:81]([O-:83])=[O:82].[K+:38]

Komponenten

15

Edukt 11

HCID 876(+-)-MethionineHCID 6032KanamycinHCID 6202Thiamine HydrochlorideHCID 10112Calcium CarbonateHCID 24843Magnesium Sulfate HeptahydrateHCID 61365Coomassie Brilliant Blue RHCID 107526(2R,3S,4R,5R)-2,3,4,5,6-pentahydroxyhexanalHCID 171548BiotinHCID 516951Monobasic potassium phosphateHCID 3423265Sodium Lauryl SulfateHCID 6097028Ammonium Sulfate

Produkt 1

HCID 23664709Penicillin G Potassium

Reagenzien, Katalysatoren, Lösungsmittel 3

HCID 62662Ferrous sulfate heptahydrateKatalysatorHCID 22138077Manganese II sulfate pentahydrateKatalysatorHCID 962WaterLösungsmittel

Versuchsvorschrift

The secretory expression system of the protransglutaminase using C. glutamicum has already been reported (WO01/23591). Then, by using the plasmid vector pPKSPTG1 for secretory expression of protransglutaminase described in WO01/23591, each of the YDK010 strain and the YDK010ΔPBP1a strain was transformed. Each of the obtained transformants was cultured in the MM liquid medium (120 g of glucose, 3 g of magnesium sulfate heptahydrate, 30 g of ammonium sulfate, 1.5 g of potassium dihydrogenphosphate, 0.03 g of iron sulfate heptahydrate, 0.03 g of manganese sulfate pentahydrate, 450 μg of thiamine hydrochloride, 450 μg of biotin, 0.15 g of DL-methionine, and 50 g of calcium carbonate in a volume of 1 L with water, adjusted to pH 7.0) containing 25 mg/l of kanamycin at 30° C. for 72 hours. After completion of the culture, culture supernatant obtained by centrifuging each culture broth was subjected to reduced SDS-PAGE, and then stained with CBB R250 (produced by Bio-Rad). Secretion amounts of the protransglutaminase were determined according to the previous report (Protein Expr. Purif., 26:329-335), and the amounts were compared. As a result, the secretion amount of the protransglutaminase was significantly improved in the YDK010ΔPBP1a strain compared with that observed for the parent strain YDK010 (FIG. 8).

Aufarbeitung

  1. customAfter completion of the culture, culture supernatant obtained

Quellen und Nachweise

ORD-Eintrag
ord-f2651ff79f8b485785e6afc1ce418be4
Quelldatensatz
uspto-grants-2015_11
Patentnummer
US09187554B2

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