تفاعل #672948

ord-da99b7641dac4d0382cb967f32db3301

معادلة التفاعل

N[C@@H](Cc1ccc(O)cc1)C(=O)O
tyrosine
Cc1ncc(COP(=O)(O)O)c(C=O)c1O
PLP
O=C(O)CN(CCN(CC(=O)O)CC(=O)O)CC(=O)O
EDTA
N[C@@H](Cc1ccc(O)cc1)C(=O)O.Oc1ccccc1
tyrosine phenol

المتفاعلات

الكواشف

لا شيء

المذيبات

ظروف التفاعل

الظروف التفصيلية
See reaction.notes.procedure_details.

المعالجة

  1. 1
    أخرىThe plasmids pTK919 [pSC101 ori, bla::tet, tyrR] and pTK922 [pSC101 ori, bla::tet, tyrR V67A, Y72C, E201G] obtained
  2. 2
    workup.ADDITIONabove were introduced into the YG17 strain
  3. 3
    أخرىto obtain the YG38 strain
  4. 4
    workup.ADDITIONThe suspension containing the disrupted microbial cells
  5. 5
    أخرىwas dialyzed against the buffer overnight

الإجراء التجريبي

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.

المصدر

DOI: 10.6084/m9.figshare.5104873.v1براءة الاختراع: US06933365B2uspto-grants-2005_08