Hydrochloric Acid → HCID 178508913, compound

HRID 1954187Notes

Reaction equation

HRID 1954187 reaction equation
[CH2:1]1[N:13]([C:14]2[CH:31]=[CH:30][C:17]([C:18](=[O:29])[NH:19][C@H:20]([C:26]([OH:28])=[O:27])[CH2:21][CH2:22][C:23]([O-:25])=[O:24])=[CH:16][CH:15]=2)[CH2:12][CH:11]2[N:2]1[C:3]1[C:4](=[O:33])[NH:5][C:6]([NH2:32])=[N:7][C:8]=1[NH:9][CH2:10]2.C1NC2NC(N)=NC(=O)C=2N2[C@H]1CN(C1C=CC(C(NC(C(O)=O)CCC(O)=O)=O)=CC=1)C2.C1C=[N+]([C@@H]2O[C@H](COP(OP(OC[C@H]3O[C@@H](N4C5N=CN=C(N)C=5N=C4)[C@H](OP(O)(O)=O)[C@@H]3O)(O)=O)(O)=O)[C@@H](O)[C@H]2O)C=C(C(N)=O)C=1.C1COCC1.C=O.C1C=[N+]([C@@H]2O[C@H](COP(OP(OC[C@H]3O[C@@H](N4C5N=CN=C(N)C=5N=C4)[C@H](OP(O)(O)=O)[C@@H]3O)(O)=O)(O)=O)[C@@H](O)[C@H]2O)C=C(C(N)=O)C=1.Cl>O>[CH2:10]1[NH:9][C:8]2[NH:7][C:6]([NH2:32])=[N:5][C:4](=[O:33])[C:3]=2[N+:2]2[C@H:11]1[CH2:12][N:13]([C:14]1[CH:31]=[CH:30][C:17]([C:18]([NH:19][C@H:20]([C:26]([O-:28])=[O:27])[CH2:21][CH2:22][C:23]([OH:25])=[O:24])=[O:29])=[CH:16][CH:15]=1)[CH:1]=2

Participants

8

Reactant 6

HCID 313Hydrochloric AcidHCID 712FormaldehydeHCID 5886NADP nicotinamide-adenine-dinucleotide phosphateHCID 8028TetrahydrofuranHCID 137182958HCID 137182958, compoundHCID 178513668HCID 178513668, compound

Product 1

HCID 178508913HCID 178508913, compound

Reagents, catalysts & solvents 1

HCID 962WaterSolvent

Procedure

5,10-Methylenetetrahydrofolate-dehydrogenase catalyzes the reaction, 5,10-methylene-THF+NADP+ →5,10-methenyl-THF+NADPH+H+. A 0.95 mL solution containing 1 μmole THF, 10 μmoles formaldehyde (fresh), 0.6 μmoles NADP and 25 μmoles n-[2-hydroxyethyl]piperazine-N'-[ethanesulfonic acid] was pre-incubated for 2 min at 30° C. Enzyme in the extraction buffer (50 μL) was then added. After the incubation continued for 15 min at 30° C., 2 mL 1 N HCl was added to stop the reaction. After 15 min at room temperature, the absorbency was measured at 355 nm. A blank containing water was used. The 5,10-methenyl-THF formed was determined using an extinction coefficient of 24,900 M-1 cm-1.

Workup

  1. additionEnzyme in the extraction buffer (50 μL) was then added
  2. waitAfter the incubation continued for 15 min at 30° C.
  3. customthe reaction
  4. waitAfter 15 min at room temperature

Sources & evidence

ORD record
ord-96756e3f96f446e2b930fecc8313f3a7
Source dataset
uspto-grants-2000_09
Patent number
US06121195

Reaction notes

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