Part:BBa_K2969025

PheOH

PheOH, Phenyalanine hydroxylase, is an enzyme which can transform Phe into Tyr. Phenylalanine hydroxylase is an enzyme that catalyzes the hydroxylation of the aromatic side-chain of phenylalanine to generate tyrosine. It is one of three members of the biopterin-dependent aromatic amino acid hydroxylases, a class of monooxygenase that uses tetrahydrobiopterin (BH4, a pteridine cofactor) and a non-heme iron for catalysis. During the reaction, molecular oxygen is heterolytically cleaved with sequential incorporation of one oxygen atom into BH4 and phenylalanine substrate

Characterization

To get the kinetic parameters of PheOH, we used BL21(DE3) strain to express PheOH with His tag and then purified PheOH with high concentration. Then we used PheOH to catalyse phenylalanine and observed the absorbance change at 275nm within 50min (Figure 1). At the same time, as shown in Figure 2, we tested the standard curve of tyrosine. After processing the data, we got the kinetic analysis curve of PheOH, which is shown in Figure 3.

Figure 1:The absorbance change at 275nm within 50min using PheOH to catalyse phenylalanine to tyrosine

Figure 2:The standard curve of tyrosine

Figure 3:Kinetic analysis of PheOH

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]