Formate → 无水磷酸二氢钾

HRID 7345

反应方程式

HRID 7345 反应方程式
C1C=[N+]([C@@H]2O[C@H](C[O:13][P:14]([O:17]P(OC[C@H]3O[C@@H](N4C5N=CN=C(N)C=5N=C4)[C@H](O)[C@@H]3O)(O)=O)([O-:16])=[O:15])[C@@H](O)[C@H]2O)C=C(C(N)=O)C=1.[CH:45]1[N:46]=[C:47]([NH2:88])[C:48]2[N:53]=[CH:52][N:51]([C@@H:54]3[O:58][C@H:57]([CH2:59][O:60][P:61]([O:64][P:65]([O:68][CH2:69][C@H:70]4[O:74][C@@H:73]([N:75]5[CH:80]=[C:79]([C:81]([NH2:83])=[O:82])[CH2:78][CH:77]=[CH:76]5)[C@H:72]([OH:84])[C@@H:71]4[OH:85])([OH:67])=[O:66])([OH:63])=[O:62])[C@@H:56]([OH:86])[C@H:55]3[OH:87])[C:49]=2[N:50]=1.C([O-])=O.C([O-])=O.[Na+].OP([O-])(O)=O.[K+]>O.C(#N)C>[P:14]([O-:17])([O-:16])([O-:15])=[O:13].[CH:45]1[N:46]=[C:47]([NH2:88])[C:48]2[N:53]=[CH:52][N:51]([C@@H:54]3[O:58][C@H:57]([CH2:59][O:60][P:61]([O:64][P:65]([O:68][CH2:69][C@H:70]4[O:74][C@@H:73]([N:75]5[CH:80]=[C:79]([C:81]([NH2:83])=[O:82])[CH2:78][CH:77]=[CH:76]5)[C@H:72]([OH:84])[C@@H:71]4[OH:85])([OH:67])=[O:66])([OH:63])=[O:62])[C@@H:56]([OH:86])[C@H:55]3[OH:87])[C:49]=2[N:50]=1

参与物

9

反应物 5

HCID 283FormateHCID 5892烟酰胺腺嘌呤二核苷酸HCID 439153辅酶NADH4 次HCID 516951磷酸钾2 次HCID 2723810甲酸钠2 次

产物 2

HCID 1061无水磷酸二氢钾HCID 439153辅酶NADH

试剂、催化剂与溶剂 2

HCID 962聚[6-[4-(4-氰基苯基)苯氧基]己基甲基丙烯酸酯]溶剂HCID 6342乙腈溶剂

实验过程

Reagents: Lyophilized nicotinamide adenine dinucleotide-diaphorase (NAD-diaphorase), p-iodonitortetrazolium violet (INT), formate dehydrogenase, and sodium formate are all obtained from Sigma Chemical Co. Acetonitrile, Na2HPO4, and KH2PO4 are of analytic grade, and reverse osmosis-deionized water is used. Phosphate buffer (100 mmol/L, pH 6.0) is prepared by mixing 100 mmol/L KH2PO4 and 100 mmol/L Na2HPO4 (5:1). Buffered NAD-diaphorase is prepared by adding 150 ml of buffer to one bottle of lyophyhlized NAD-diaphorase (Sigma). Buffered NAD-diaphorase-INT is prepared by addition of 140 mg INT (Sigma) to 140 ml of buffered NAD-diaphorase, and recombinant P. pastoris FDH is lyophilized. Specific activity of the P. pastoris FDH is 0.4 to 1.0 unites/mg solid (0.4 to 1.8 units/mg protein). Portions (5 to 15 mg) are weighed into 1.5 ml glass vials and stored at −70° C. One vial is used for each batch analysis. Each is reconstituted with about 100 to 200 μl/mg buffered NAD-diaphorase (5° C.) and kept in an ice bath during analysis. Serum and aqueous standards (10 ml each) are supplemented with stock aqueous sodium formate solutions containing 10.0 or 1.0 g/L to produce concentrations ranging from 0 to 400 mg/L with less than 1% volumetric alteration of the matrix. Working reagent and final reaction (cuvette) concentrations are shown in the table, below.

后处理

  1. customBuffered NAD-diaphorase is prepared
  2. customstored at −70° C
  3. customkept in an ice bath during analysis
  4. additioncontaining 10.0 or 1.0 g/L
  5. customto produce
  6. concentrationconcentrations
  7. customWorking reagent and final reaction (cuvette) concentrations

来源与证据

ORD记录
ord-75aa97babaad4e7680f7370acd01a5be
来源数据集
uspto-grants-2006_08
专利号
US07087418B2

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