Penicillin G → Penicillin G Potassium
Reaction equation
OP([O-])([O-])=O.[K+:6].[K+].OC(CCCC[C@H]1[C@@H]2[C@@H](NC(N2)=O)CS1)=O.[CH3:24][C:25]1([CH3:46])[S:29][C@@H:28]2[C@H:30]([NH:33][C:34]([CH2:36][C:37]3[CH:38]=[CH:39][CH:40]=[CH:41][CH:42]=3)=[O:35])[C:31](=[O:32])[N:27]2[C@H:26]1[C:43]([OH:45])=[O:44]>>[CH3:24][C:25]1([CH3:46])[S:29][C@@H:28]2[C@H:30]([NH:33][C:34]([CH2:36][C:37]3[CH:38]=[CH:39][CH:40]=[CH:41][CH:42]=3)=[O:35])[C:31](=[O:32])[N:27]2[C@H:26]1[C:43]([O-:45])=[O:44].[K+:6]Participants
Conditions
- Temperature
- 30 °C
Procedure
Culture treated as under A is incubated for about 72 seconds at 30° C. in the shaking machine until it attains a cell titre of >109 per ml of culture solution. 0.1 ml of this cell suspension is transferred into 10 ml of the following nutrient solution (nutrient solution B): 0.05% NaH2PO4, 0.20% K2HPO4, 0.05% MgSO4.7H2O, 0.02% CaCl2.2H2O, 0.005% MnSO4.4H2O, 0.005% (Fe)2SO4.7H2O, 0.10% (NH4)SO4, 0.0001% biotine, 0.10% Tween 80 and 0.10% BNC. After shaking for 18 hours at 30° C. diluted with fresh, preheated nutrient solution of the same composition to about 106 cells per ml, 1000 IU penicillin G are added and incubation for a further 7 hours is carried out. The antibioticum is then removed by centrifuging of the cells with washing with sterile, physiologic sodium chloride solution and the cells further incubated for 3 days in the above-mentioned medium with 0.2% sitosterol instead of BNC. The penicillin treatment is repeated once more and the cell suspension finally plated out on the same nutrient medium A plus 1.6% agar. After the propagation of the cells to visible colonies, the cells are transfered with the aid of the "replica stamp technique" on nutrient medium B plus 1.6% agar and those colonies are selected and used as strain cultures which are not able to continue to grow, or scarecly able, on this medium.
Workup
- additionCulture treated as under
- additionare added
- waitincubation for a further 7 hours is carried out
- customThe antibioticum is then removed by centrifuging of the cells
- washwith washing with sterile, physiologic sodium chloride solution
- waitthe cells further incubated for 3 days in the above-mentioned medium with 0.2% sitosterol instead of BNC
Sources & evidence
- ORD record
- ord-fd999ae844b44b699edb1eb9f77da690
- Source dataset
- uspto-grants-1982_12
- Patent number
- US04362815
Reaction notes
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