Nadide (JAN/USAN/INN) → Nadh

HRID 634075

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HRID 634075 반응식
NCC(O)=O.[OH-].[K+].[O-]S([O-])(=O)=O.[Mg+2].[CH:14]1[CH:19]=[N+:18]([C@@H:20]2[O:24][C@H:23]([CH2:25][O:26][P:27]([O:30][P:31]([O:34][CH2:35][C@H:36]3[O:40][C@@H:39]([N:41]4[C:45]5[N:46]=[CH:47][N:48]=[C:49]([NH2:50])[C:44]=5[N:43]=[CH:42]4)[C@H:38]([OH:51])[C@@H:37]3[OH:52])([OH:33])=[O:32])([OH:29])=[O:28])[C@@H:22]([OH:53])[C@H:21]2[OH:54])[CH:17]=[C:16]([C:55]([NH2:57])=[O:56])[CH:15]=1>>[CH:47]1[N:48]=[C:49]([NH2:50])[C:44]2[N:43]=[CH:42][N:41]([C@@H:39]3[O:40][C@H:36]([CH2:35][O:34][P:31]([O:30][P:27]([O:26][CH2:25][C@H:23]4[O:24][C@@H:20]([N:18]5[CH:17]=[C:16]([C:55]([NH2:57])=[O:56])[CH2:15][CH:14]=[CH:19]5)[C@H:21]([OH:54])[C@@H:22]4[OH:53])([OH:29])=[O:28])([OH:33])=[O:32])[C@@H:37]([OH:52])[C@H:38]3[OH:51])[C:45]=2[N:46]=1

참가 물질

4

반응물 3

HCID 5893Nadide (JAN/USAN/INN)HCID 24083Sulfuric acid magnesium salt (1:1)HCID 87772558Glycine KOH

생성물 1

HCID 439153Nadh

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온도
45 °C

실험 절차

Alcohol dehydrogenase activities were measured spectrophotometrically essentially as described previously (Hektor, Kloosterman et al. (2002) Chem 277(49): 46966-46973) and the reaction mixture contained: 100 mM Glycine-KOH pH 9.5 (unless otherwise stated), 5 mM MgSO4, 0.5 mM NAD+ and 500 mM alcohol (methanol, ethanol, propanol, 1,3-propanediol, or butanol). NAD+ was substituted with equal concentrations of NADP+, FMN+, and FAD+, when indicated. The reaction mixture for measurements of formaldehyde reductase activity contained: 50 mM Potassium-phosphate buffer pH=6.7, 0.15 mM NADH, 1 mM DTT and 11.6 mM (0.1-116 mM) formaldehyde. The assay components were mixed in the cuvette and pre-warmed to 45° C., unless otherwise stated. The reactions were started by addition of 5-40 μg of the purified MDH proteins, and the production of NADH was monitored at 340 nm for 4 minutes. One unit of MDH activity was defined as the amount of enzyme needed to produce 1 μmol NADH per minute under the conditions described above. Purified Act (0.1-40 μg) or NudF (20 μg) proteins were added to the reaction mixtures as indicated in the text.

후처리

  1. concentrationNAD+ was substituted with equal concentrations of NADP+, FMN+, and FAD+, when
  2. additionThe assay components were mixed in the cuvette
  3. additionThe reactions were started by addition of 5-40 μg of the purified MDH proteins

출처 및 근거

ORD 기록
ord-18ad074c4faa4d3a967e111daf0464c9
출처 데이터셋
uspto-grants-2016_04
특허 번호
US09322000B2

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