Reaction equation
CC1(C)S[C@@H]2[C@H](NC(C(C(O)=O)C3C=CC=CC=3)=O)C(=[O:9])N2[C@H]1C(O)=O.CC(S[C@@H:31]1[O:36][C@H:35]([CH2:37][OH:38])[C@H:34]([OH:39])[C@H:33]([OH:40])[C@H:32]1[OH:41])C.[Mg+2].[Cl-].[Cl-].[Na+].[Cl-].C([O-])([O-])=O.[Ca+2]>>[OH:39][C:34]1[C@@H:33]([C@@H:32]([OH:41])[CH2:31][OH:36])[O:40][C:37](=[O:38])[C:35]=1[OH:9]Participants
Procedure
Five ml of one overnight culture of E. coli BL21 (DE3)/pET21b-SNDHaiHis in LB with carbenicillin 50 μg/ml was inoculated into 200 ml of the same medium. The cells were cultivated at 230 rpm at 37° C. for 2 h, then induced with 1 mM IPTG and continued to be cultivated at 230 rpm at 25° C. for 3 h. The resulting culture was centrifuged and washed twice with saline and the cell pellet was resuspended in 2 ml of water. The cells were used for resting cell reaction with the reaction mixture (500 in 5 ml tube) containing cells at OD600=10, 1% sorbosone monohydrate, 5 μM PQQ, 5 mM MgCl2, 0.3% NaCl, and 1% CaCO3 conducted at 30° C. for 15 h. 0.14 g/L of Vitamin C was produced after incubation for 15 h. When the resting cell reaction was done with 1 μM PQQ (the other conditions were the same as those described above), 0.05 g/L of Vitamin C was produced after incubation for 3 h.
Workup
- waitto be cultivated at 230 rpm at 25° C. for 3 h
- washwashed twice with saline
- customcell reaction with the reaction mixture (500 in 5 ml tube)
- waitconducted at 30° C. for 15 h
Sources & evidence
- ORD record
- ord-c44f3f37b89f4674bbef6ada592d7a72
- Source dataset
- uspto-grants-2015_08
- Patent number
- US09115378B2
Linked notes
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