Materials and Reagents

1. Comamonas sp. 7D-2
   
2. LB medium
   
3. Substrate, 3,5-dibromo-4-hydroxybenzoate (DBHB) or 3-bromo-4-hydroxybenzoate (BHB) (Sigma-Aldrich)
   
4. Electron donor, NADPH or NADH (Sangon Biotech, catalog numbers: Y4433000-100 mg and NB0642-1 g , respectively)
   
5. Reaction inhibitor, sodium dithionite (Na₂S₂O₄) (Sigma-Aldrich, catalog number: 7775-14-6 )
   
6. Protein Quantification Kit (Sangon Biotech, catalog number: BE530-100 ml )
   
7. Phosphate buffered saline (PBS) (Sambrook and Russell, 2001) (see Recipes)
   
8. Mobile phase of HPLC (see Recipes)
   

Equipment

1. 7 ml centrifuge tube
   
2. Membrane filtration (pore size, 0.22 μm)
   
3. Centrifuge
   
4. HPLC (600 controller, Rheodyne 7725i manual injector and 2487 Dual λ Absorbance Detector) (Waters)
   
5. Ultrasonic instrument
   
6. Fast protein liquid chromatography
   

Procedure