Tannase is found in tannin rich plants and also in microorganisms. It can hydrolyze the ester bond and the
phenol carboxyl bond to form gallic acid and glucose.
Using propyl gallate (PG) as the substrate of tannase, it has a characteristic absorption peak at 270nn.
Tannase activity can be calculated by measuring the absorbance at 270 nm.
Required but Not Provided:
Ultraviolet spectrophotometer/microplate reader, desk centrifuge, water-bath, transferpettor, micro quartz
cuvette/96 well flat-bottom UV plate, mortar/homogenizer, ethanol, ice and distilled water.
Protocol
I. Preparation:
1. Tissue:
Add 1 mL of extraction reagent to 0.1 g of tissue. Homogenate on ice. Centrifuge at 10000 rpm 4℃ for 10
minutes. Take the supernatant on ice for test.
2. Cells or bacterial
Collect bacteria or cells into the centrifuge tube. Discard the supernatant after centrifugation. It is
suggested to take about 5 million bacteria/cell and add 1 mL extraction reagent. Bacteria/cell is split by
ultrasonication (power 20%, ultrasonic 3s, interval 10s, repeat for 30 times). Centrifuge at 10000 rpm 4℃
for 10 minutes. Take the supernatant on ice for test.
II. Determination
