How to calculate pectinase enzyme activity as, we have values of OD @540 nm obtained from DNS method of different Ph and Temp of sub merged fermentation using wheat bran and citrus peel ? please let me know the calculation of total activity and specific activity by using standard curve?
The pectinase enzyme activity can be calculated using the following formula:
Enzyme activity (U/mL) = (ΔAbs/Δt) * V * C
where:
- ΔAbs is the change in absorbance at a wavelength of 540 nm over a period of time (Δt)
- V is the final reaction volume (in mL)
- C is the concentration of the substrate (pectin) in the final reaction mixture (in mg/mL)
The absorbance is measured using a spectrophotometer. The substrate concentration is typically determined using a standard curve.
For example, if the change in absorbance is 0.100, the reaction time is 1 minute, the final reaction volume is 5 mL, and the substrate concentration is 100 mg/mL, then the enzyme activity is:
Enzyme activity (U/mL) = (0.100/1) * 5 * 100 = 50 U/mL
The units of enzyme activity are typically micromoles (µmol) of substrate converted into product per minute (per mL).
Here are the steps on how to calculate the pectinase enzyme activity:
The enzyme activity can also be expressed in other units, such as katals (kat) or international units (IU). 1 kat = 60 µmol/min. 1 IU = 1 µmol/min/mL.
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