The separation of vanillate O-demethylase from protocatechuate 3,4-oxygenase by ultracentrifugation.
1. Protocatechuate 3,4-oxygenase in the soluble part of a cell-free extract of Pseudomonas fluorescens (strain T) sedimented more rapidly than vanillate O-demethylase under specified conditions in a preparative ultracentrifuge. 2. The supernatant from this process contained vanillate O-demethylase and formaldehyde dehydrogenase, and when supplemented with NADH oxidized vanillate with an uptake of 1 mole of oxygen/mole of substrate and accumulation of protocatechuate. 3. This uptake was decreased to 0.5mole/mole of substrate in the presence of semicarbazide as trapping agent for formaldehyde. 4. Reasons are presented for the process of methyl group removal from vanillate being oxidative demethylation.
D002474:Cell-Free System; D005557:Formaldehyde; D008365:Manometry; D009243:NAD; D010084:Oxidation-Reduction; D010105:Oxygenases; D011549:Pseudomonas / Q000201:enzymology*; D014166:Transferases; D014461:Ultracentrifugation; D014465:Ultrasonics
D009243:NAD; D005557:Formaldehyde; D010105:Oxygenases; D014166:Transferases