(+-)-Methionine → Penicillin G Potassium

HRID 716431

反応式

HRID 716431 の反応式
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]

参加物

15

反応物 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

生成物 1

HCID 23664709Penicillin G Potassium

試薬・触媒・溶媒 3

HCID 62662Ferrous sulfate heptahydrate触媒HCID 22138077Manganese II sulfate pentahydrate触媒HCID 962Water溶媒

実験手順

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).

後処理

  1. customAfter completion of the culture, culture supernatant obtained

出典と根拠

ORDレコード
ord-f2651ff79f8b485785e6afc1ce418be4
出典データセット
uspto-grants-2015_11
特許番号
US09187554B2

関連ノート

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