Reaction equation
CC1(C)S[C@@H]2[C@H](NC([C@H](N)C3C=CC=CC=3)=O)C(=O)N2[C@H]1C(O)=O.CC(S[C@@H]1O[C@H](CO)[C@H](O)[C@H](O)[C@H]1O)C.C([NH:43][CH:44]([C:55]([OH:57])=[O:56])[CH2:45][C:46]1[C:54]2[C:49](=[CH:50][CH:51]=[CH:52][CH:53]=2)[NH:48][CH:47]=1)(=O)C.[C:58]([NH:61][C@H:62]([C:73]([OH:75])=[O:74])[CH2:63][C:64]1[C:72]2[C:67](=[CH:68][CH:69]=[CH:70][CH:71]=2)[NH:66][CH:65]=1)(=[O:60])[CH3:59]>CCCCCCCCCCCCCCCC[N+]1C=CC=CC=1.[Br-]>[NH2:43][C@@H:44]([C:55]([OH:57])=[O:56])[CH2:45][C:46]1[C:54]2[C:49](=[CH:50][CH:51]=[CH:52][CH:53]=2)[NH:48][CH:47]=1.[C:58]([NH:61][C@@H:62]([C:73]([OH:75])=[O:74])[CH2:63][C:64]1[C:72]2[C:67](=[CH:68][CH:69]=[CH:70][CH:71]=2)[NH:66][CH:65]=1)(=[O:60])[CH3:59]Participants
Procedure
Cells of the E. coli transformant were inoculated on an LB agar plate. Each colony grown on the plate was inoculated with a toothpick to a well of a microtiter plate which contained 150 μl of LB culture medium containing 50 μg/ml ampicillin and 0.02 mM IPTG, and then the microtiter plate was covered with Breathable Sealing Membrane (Nalgene). The microtiter plate was incubated at 37° C. for 18 hours while being shaken. 10 μl of the culture medium solution was added to the reaction solution containing 300 mM Tris-hydrochloride buffer (pH 8.0), 0.1% cetylpyridinium bromide, 5% N-acetyl-DL-tryptophan, 1% N-acetyl-L-tryptophan, and distilled water in the wells of a microtiter plate. The plate was incubated at 25° C. for 1 hour while being shaken. Then, D-tryptophan produced from N-acetyl-D-tryptophan was quantified by colorimetry according to the TNBS method.
Workup
- addition10 μl of the culture medium solution was added to the reaction solution
- distillationdistilled water in the wells of a microtiter plate
- waitThe plate was incubated at 25° C. for 1 hour
- stirringwhile being shaken
Sources & evidence
- ORD record
- ord-5b0d6c2848b6414cb8c8f68cb3cd146e
- Source dataset
- uspto-grants-2007_10
- Patent number
- US07282356B2
Linked notes
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