反応 #672948

反応方程式

tyrosine

PLP

EDTA

→

tyrosine phenol

溶媒

potassium phosphate

反応条件

後処理

1. 1
   その他The 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
   その他to obtain the YG38 strain
4. 4
   workup.ADDITIONThe suspension containing the disrupted microbial cells
5. 5
   その他was dialyzed against the buffer overnight

実験手順

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.

出典