N-Acetyltryptophan → D-Tryptophan

HRID 1180470

반응식

HRID 1180470 반응식
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]

참가 물질

7

반응물 4

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

생성물 2

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

시약·촉매·용매 1

HCID 8816Cetylpyridinium Bromide촉매

실험 절차

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.

후처리

  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

출처 및 근거

ORD 기록
ord-a144e3511a7f48afba62cb43f9a36150
출처 데이터셋
uspto-grants-2007_10
특허 번호
US07282356B2

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