Main reagents:

Yuchuang phenol (analytical grade), succinic acid, NaOH standard solution.

Substrate preparation: Mix 137.4 μ l of guaiacol (analytical grade) with 4.5g of succinic acid, adjust the pH to 4.5 with NaOH solution, and dilute to 1L with distilled water.

Main instruments:

⑴ UV-2100 UV visible spectrophotometer;

⑵ DELTA 320 pH meter.

Test method:

Dilute the enzyme solution to the required multiple.

Take 2.4ml of substrate, add 0.6ml of enzyme solution, mix well, and react at 30 degrees for 30 minutes.

Measure its absorbance at 465 nanometers.

Calculation formula:

Enzyme activity (U/L)=A × n × 13.89

A absorbance value n dilution factor 13.89 coefficient

definition:

The enzyme required to oxidize 1 μ mol of guaiacol within one minute is one enzyme activity unit (U).