Hi All,
I want to calculate the L-asparaginase enzyme activity. I ain't sure which formula should I use. If I use the standard one [1 unit equalling the amount of L-asparaginase that liberates 1 umol of ammonia from L-asparagine per min at pH 8.6 and 37degC (Yellin and Wriston, 1966)], my values are very less.
If I calculate the amount of ammonia released from standard graph then by applying it in
units/ ml enzyme = umol of ammonia released * Total reaction Volume
Volume of filtrate * Incubation time
results in 4 digit value, I do not know which one should I take in account. Expecting suggestions and help from you all.
Thanks in advance.
Dear Yogi,
For the calculation of the enzyme activity, I suggest that you use the following equation:
Enzyme Activity (uml) =
Amount of NH4+ liberated / (incubation time x ml of enzyme taken for a test).
For an example, please see attached file.
Hoping this will be helpful,
Rafik
In your equation, you have multiplied by time instead of divided by it. Also, to calculate units/ml of enzyme, the ml of enzyme in question should be the volume of the original sample (in ml) that went into the reaction. For example, if you performed the reaction in a 1-ml volume, and you added 10 µl of your enzyme sample into the reaction, then your should divide (not multiply) by 0.01 ml.
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