Expression of mouse tyrosine hydroxylase in Escherichia coli.
Enzymatically active mouse tyrosine hydroxylase (TH) was successfully expressed at a high level in Escherichia coli using a T7 RNA polymerase directed expression system. The specific activity of mouse TH in E. coli cell lysate was 7.5 nmol/mg protein/min. Kinetic characteristics of recombinant TH were examined. Km for tyrosine and (6R)-tetrahydrobiopterin (6R-BH4) cofactor were determined to be 7.2 microM (420 microM 6R-BH4), 19 microM [( 6R-BH4] less than 55 microM, 20 microM tyrosine) and 54 microM [( 6R-BH4] greater than 55 microM, 20 microM tyrosine), respectively. These were in good agreement with previously reported values for this enzyme.
Department of Biochemistry, Nagoya University School of Medicine, Japan.
S
S
Ichikawa
S
S
Nasrin
T
T
Nagatsu
D000818:Animals; D001483:Base Sequence; D015153:Blotting, Western; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics*; D005822:Genetic Vectors; D007700:Kinetics; D051379:Mice; D008969:Molecular Sequence Data; D008970:Molecular Weight; D015345:Oligonucleotide Probes; D010957:Plasmids; D016133:Polymerase Chain Reaction / Q000379:methods; D011994:Recombinant Proteins / Q000302:isolation & purification / Q000378:metabolism; D015183:Restriction Mapping; D014446:Tyrosine 3-Monooxygenase / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism
D015345:Oligonucleotide Probes; D011994:Recombinant Proteins; D014446:Tyrosine 3-Monooxygenase