Determination of peroxidase activity (color ratio)

the determination of peroxidase activity (color ratio)principle peroxidase is widely distributed in various
tissues and
organs of plants. In the presence of hydrogen oxide, peroxidase can make the more invasive wood phenol oxidation, the production of tea brown matter, can be used
hydrolumometer
to measure the content of the result.。 Instrument pharmaceutical 721 phosphorescometer
centrifuge sheet
balanceresearch magnet force
agitator30% hydrogen peroxide 20mmol/LKH2 PO4 100mmol/L phosphoric acid buffer, pH6.0 (see Schedule 2 ) Reaction mixture: 100mmol/L phosphate buffer (pH6.0) 50ml in a
beech
, add the more invasive wood phenol 28 μl,
heat
on the magnetic mixer stirring, until the more invasive wood phenol dissolves, after the solution is cooled, add 30% hydrogen peroxide 19 sl, mix evenly, store in the refrigerator.。 Step 1. 1. Called plant material 1g, plus 20mmol/L KH2 PO4 5ml, ground into a homogenous slurry in the research, with 4000r/min centrifugal 15 minutes, pouring out the liquid stored in the cold, residue with 5 ml KH2 PO4 solution extracted once, combined twice on the liquid, stored in the cold spare.2. Take the optical diameter 1cm color cup 2, add the reaction mixture 3ml, KH2 PO4 1ml in one, as a zero control, and add the reaction mixture 3ml in the other, the above enzyme solution 1 ml (e.g. enzyme activity is too high to dilute), immediately turn on the stopwatch recording time, measure the absorbance value on the hydrometry meter, read once every 1 minute, read at a wavelength of 470nm.3. The enzyme activity size is expressed in the value of the light absorption per minute, i.e. in the
protein
(or fresh weight g). Protein content determination can be found in Experiment 56.。