NADPH → HCID 178434963, 화합물

HRID 1794979

반응식

HRID 1794979 반응식
[CH3:1][C@H:2]1[O:8][C@H:7]([O:9][P:10]([O:13][P:14]([O:17][CH2:18][C@H:19]2[O:23][C@@H:22]([N:24]3[C:28]4[NH:29][C:30]([NH2:34])=[N:31][C:32](=[O:33])[C:27]=4[N:26]=[CH:25]3)[C@H:21]([OH:35])[C@@H:20]2[OH:36])([OH:16])=[O:15])([OH:12])=[O:11])[C@@H:6]([OH:37])[C@@H:5]([OH:38])[C:3]1=[O:4].C1N([C@@H]2O[C@H](COP(OP(O[C@H]3O[C@H](CO)[C@@H](O)[C@H](O)[C@@H]3O)(O)=O)(O)=O)[C@@H](O)[C@H]2O)C2NC(N)=NC(=O)C=2N=1.O=C[C@@H]([C@H]([C@@H]([C@@H](CO)O)O)O)O.[Mg+2].[Cl-].[Cl-].C1N=C(N)C2N=CN([C@@H]3O[C@H](COP(OP(OC[C@H]4O[C@@H](N5C=C(C(N)=O)CC=C5)[C@H](O)[C@@H]4O)(O)=O)(O)=O)[C@@H](O)[C@H]3OP(O)(O)=O)C=2N=1>O>[CH3:1][CH:2]1[O:8][CH:7]([O:9][P:10]([O:13][P:14]([O:17][CH2:18][CH:19]2[O:23][CH:22]([N:24]3[C:28]4[NH:29][C:30]([NH2:34])=[N:31][C:32](=[O:33])[C:27]=4[N:26]=[CH:25]3)[CH:21]([OH:35])[CH:20]2[OH:36])([OH:16])=[O:15])([OH:12])=[O:11])[CH:6]([OH:37])[CH:5]([OH:38])[CH:3]1[OH:4]

참가 물질

7

반응물 5

HCID 5884NADPHHCID 107526(2R,3S,4R,5R)-2,3,4,5,6-pentahydroxyhexanal2회HCID 5360315Magnesium ChlorideHCID 178461617HCID 178461617, 화합물2회HCID 178508932HCID 178508932, 화합물3회

생성물 1

HCID 178434963HCID 178434963, 화합물

시약·촉매·용매 1

HCID 962Water용매

실험 절차

A coupled assay was employed to assay YEF B activity. This was required since its substrate, GDP-4-keto-6-deoxymannose, is unstable and not commercially available. Twenty mUnit of GDP-mannose dehydratase was allowed to react with 480 nMole of GDP-mannose for 20 minutes under standard assay conditions as previously. At this time greater than 90% of the GDP-mannose has been converted to GDP-4-keto-6-deoxymannose, the substrate for YEF B. The following additions were then made: 500 nmole glucose, MgCl2 to 20 mM, NADPH to 0.15 mM, 10 mUnit glucose dehydrogenase, 100 nmole CTP (to normalize for losses that may occur when sample is injected onto HPLC), YEF B and water to 50 μl. The reaction was allowed to proceed for ten minutes at which point the reaction was stopped by freezing on dry ice. After thawing 10 μl aliquots were analyzed by reverse phase HPLC analysis. The amount of GDP fucose formed was calculated by analyzing a separate assay mix in the absence of GDP-mannose dehydratase to yield a 100% value for the absorption peak of GDP-mannose. This peak area was then divided by the peak area of GDP-fucose formed to yield percentage of GDP-fucose formed. (% GDP-fucose (nMole GDP-mannose in reaction)=nMole GDP-fucose formed.

후처리

  1. waitto proceed for ten minutes at which

출처 및 근거

ORD 기록
ord-26994cbeecf54f648a5a9f8a406dadb3
출처 데이터셋
uspto-grants-2006_04
특허 번호
US07026142B2

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