Water → Coenzyme A

HRID 444669

反応式

HRID 444669 の反応式
C1(=O)NC(=O)C=C1.OC(CCCC[C@H]1[C@@H]2[C@@H](NC(N2)=O)CS1)=O.[Na].[Na].[C@@H:26]1([N:64]2[C:73]3[N:72]=[CH:71][N:70]=[C:68]([NH2:69])[C:67]=3[N:66]=[CH:65]2)[O:63][C@H:35]([CH2:36][O:37][P:38]([O:41][P:42]([O:45][CH2:46][C:47]([C@H:50]([C:52]([NH:54][CH2:55][CH2:56][C:57]([NH:59][CH2:60][CH2:61][SH:62])=[O:58])=[O:53])[OH:51])([CH3:49])[CH3:48])([OH:44])=[O:43])([OH:40])=[O:39])[C@@H:29]([O:30][P:31]([OH:34])([OH:33])=[O:32])[C@H:27]1[OH:28].O.CC([C@@H](O)C(NCCC(NCCS)=O)=O)(COP(OP(OC[C@H]1O[C@@H](N2C3N=CN=C(N)C=3N=C2)[C@H](O)[C@@H]1OP(O)(O)=O)(O)=O)(O)=O)C.OC(CCCC[C@H]1[C@@H]2[C@@H](NC(N2)=O)CS1)=O>CN(C=O)C.CC#N.O.CC#N>[CH3:49][C:47]([C@@H:50]([OH:51])[C:52]([NH:54][CH2:55][CH2:56][C:57]([NH:59][CH2:60][CH2:61][SH:62])=[O:58])=[O:53])([CH2:46][O:45][P:42]([O:41][P:38]([O:37][CH2:36][C@H:35]1[O:63][C@@H:26]([N:64]2[C:73]3[N:72]=[CH:71][N:70]=[C:68]([NH2:69])[C:67]=3[N:66]=[CH:65]2)[C@H:27]([OH:28])[C@@H:29]1[O:30][P:31]([OH:34])([OH:33])=[O:32])([OH:40])=[O:39])([OH:44])=[O:43])[CH3:48]

参加物

8

反応物 4

HCID 962WaterHCID 22896102Maleimide-biotinHCID 86643215coenzyme A disodium saltHCID 86643216CoA biotin

生成物 1

HCID 87642Coenzyme A

試薬・触媒・溶媒 3

HCID 6228Dimethylformamide溶媒2 回HCID 6342Acetonitrile溶媒2 回HCID 18373917Acetonitrile--water (1/1)溶媒

実験手順

To a solution of biotin-maleimide 1 (1 mg, 0.0022 mmol) in 100 μl DMF, a solution of coenzyme A disodium salt (1.79 mg, 0.0022 mmol, 1 eq.) in 90 μl DMF and 10 μl 50 mM Tris-Cl pH 7.5 is added. The mixture is stirred for 4 hours at room temperature. It is then diluted with CH3CN/H2O 1:4, and aliquots of 500 μl are injected on a preparative HPLC column: Gradient (A=H2O 99%, CH3CN 1%, 50 mM NH4OAc/B=CH3CN) from A/B 95:5 to A/B 80:20 in 2 min, to A/B 68:32 in 7 min, to A/B 20:80 in 2 min, then back to A/B 95:5. The retention time of the CoA-biotin is 6.5 min. Fractions containing the desired product are concentrated in vacuo, dissolved in DMSO, and an analytical amount injected to control the purity. The pure fractions are combined. The concentration of CoA-biotin is determined by absorption at 260 nm (ε(adenine, 260 nm)=15'300 [M−1 cm−1]). The yield of CoA-Bt is 0.895 mg (33%).

後処理

  1. waitto A/B 68:32 in 7 min
  2. waitto A/B 20:80 in 2 min

出典と根拠

ORDレコード
ord-89a59de69fc74c1a972f8cc3068a1e93
出典データセット
uspto-grants-2013_07
特許番号
US08476031B2

関連ノート

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