Pyridoxal 5'-phosphate → Tyrosine phenol

HRID 585766Notes

Reaction equation

HRID 585766 reaction equation
[NH2:1][C@H:2]([C:11]([OH:13])=[O:12])[CH2:3][C:4]1[CH:9]=[CH:8][C:7]([OH:10])=[CH:6][CH:5]=1.CC1C(O)=C(C=O)C(COP(O)(O)=O)=CN=1.C(N(CC(O)=O)CC(O)=O)CN(CC(O)=O)CC(O)=O>P([O-])([O-])([O-])=O.[K+].[K+].[K+]>[C:7]1([OH:10])[CH:8]=[CH:9][CH:4]=[CH:5][CH:6]=1.[NH2:1][C@H:2]([C:11]([OH:13])=[O:12])[CH2:3][C:4]1[CH:5]=[CH:6][C:7]([OH:10])=[CH:8][CH:9]=1

Participants

5

Reactant 3

HCID 1051Pyridoxal 5'-phosphateHCID 6049Ethylenediaminetetraacetic AcidHCID 6057L-Tyrosine

Product 1

HCID 67207896Tyrosine phenol

Reagents, catalysts & solvents 1

HCID 62657Potassium Phosphate, TribasicSolvent

Procedure

The plasmids pTK919 [pSC101 ori, bla::tet, tyrR] and pTK922 [pSC101 ori, bla::tet, tyrR V67A, Y72C, E201G] obtained as described above were introduced into the YG17 strain to obtain the YG38 strain and the YG40 strain, respectively. These strains were cultured overnight at 30° C. in 100 ml of the basal medium containing 0.1% tyrosine. The microbial cells were suspended in 10 mM potassium phosphate buffer (pH 7.0), 0.2 mM PLP (pyridoxal 5′-phosphate), 5 mM 2-mercaptoethnol and 4 mM EDTA (pH 7.0) and disrupted by sonication. The suspension containing the disrupted microbial cells was dialyzed against the buffer overnight. The amount of protein and tyrosine phenol lyase activity in the crude enzyme solution obtained as described above were determined.

Workup

  1. customThe plasmids pTK919 [pSC101 ori, bla::tet, tyrR] and pTK922 [pSC101 ori, bla::tet, tyrR V67A, Y72C, E201G] obtained
  2. additionabove were introduced into the YG17 strain
  3. customto obtain the YG38 strain
  4. additionThe suspension containing the disrupted microbial cells
  5. customwas dialyzed against the buffer overnight

Sources & evidence

ORD record
ord-da99b7641dac4d0382cb967f32db3301
Source dataset
uspto-grants-2005_08
Patent number
US06933365B2

Reaction notes

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