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N-Acetyltryptophan → D-Tryptophan

HRID 1180470
ノート
印刷プレビュー

反応式

HRID 1180470 の反応式
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]
参加物実験手順後処理出典と根拠反応ノート

参加物

7

反応物 4

HCID 2002N-AcetyltryptophanHCID 6249AmpicillinHCID 656894isopropyl beta-D-thiogalactopyranosideHCID 700653N-Acetyl-L-tryptophan

生成物 2

HCID 9060D-TryptophanHCID 439917N-acetyl-D-tryptophan

試薬・触媒・溶媒 1

HCID 8816
Cetylpyridinium Bromide
触媒

実験手順

Three types of the E. coli transformant libraries obtained in Example 11 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. The same reaction as described above was conducted simultaneously using wild-type E.coli strain. Then, mutants with increased activity for producing D-tryptophan compared with wild-type strain were isolated as mutants with reduced inhibition. Screening of about. 200 colonies yielded a number of mutants whose competitive inhibition had been reduced.

後処理

  1. customcoli transformant libraries obtained in Example 11
  2. addition10 μl of the culture medium solution was added to the reaction solution
  3. distillationdistilled water in the wells of a microtiter plate
  4. waitThe plate was incubated at 25° C. for 1 hour
  5. stirringwhile being shaken

出典と根拠

ORDレコード
ord-a144e3511a7f48afba62cb43f9a36150
出典データセット
uspto-grants-2007_10
DOI
10.6084/m9.figshare.5104873.v1
特許番号
US07282356B2

反応ノート

読み込み中…

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