Reaction #1379907

ord-64fd689fb2ec4abfa580540c19f4d292

Reaction equation

O=C(O)[C@@H]1CCCN1
L-proline
O=C(O)CCC(=O)C(=O)O
2-ketoglutaric acid
O=C1O[C@H]([C@@H](O)CO)C(O)=C1O
L-ascorbic acid
O=S(=O)(O)CCN1CCOCC1
MES
→
O=C(O)[C@@H]1C[C@@H](O)CN1
trans-4-hydroxy-L-proline

Reactants

O=C(O)[C@@H]1CCCN1
L-proline
O=C(O)CCC(=O)C(=O)O
2-ketoglutaric acid
O=C1O[C@H]([C@@H](O)CO)C(O)=C1O
L-ascorbic acid
—
ferrous sulfate
O=S(=O)(O)CCN1CCOCC1
MES

Reagents

None

Conditions

Temperature
100°CELSIUS
Detailed conditions
See reaction.notes.procedure_details.

Workup

  1. 1
    OtherThe enzyme preparation
  2. 2
    Otherthe reaction

Procedure

The enzyme preparation to be determined is added to 80 mM MES buffer (pH 6.5) containing 4 mM L-proline, 8 mM 2-ketoglutaric acid, 2 mM ferrous sulfate and 4 mM L-ascorbic acid to make 250 μl in total, and the mixture was allowed to stand at 30° C. for 20 minutes. The reaction mixture is heated at 100° C. for 2 minutes so as to stop the reaction, and the amount of the trans-4-hydroxy-L-proline produced in the reaction mixture is determined by high performance liquid chromatography (hereinafter referred to as HPLC).

Source

DOI: 10.6084/m9.figshare.5104873.v1Patent: US07238501B2uspto-grants-2007_07