I want to hydrolyze oat protein by using enzyme Alcalase 2.4 L ( 2.4 AU g).
how can we calculate enzyme / substrate ratio 0.3 AU g of protein??
To calculate the (molecular) enzyme/substrate ratio, you would need to know the molar concentration of your enzyme. To get this from the specific activity of the enzyme preparation, you would need the purity of your preparation and the molecular weight of your enzyme, or the specific molar activity of the pure enzyme.
IUBMB would give you the appropriate unit definitions and BRENDA is every biochemists friend, a great resource.
https://www.brenda-enzymes.org/index.php
Enzyme /Substrate ratio can be determined by using this formula:Concentration of enzyme concentration / molecular weight
Substrate concentration/ molecular weight.
Protein and substrate concentration can be determined by spectrophotometer, where the formula is
A= E x C x L
where
E= extinction coefficient ( value given in literature)
C= concentration
L =path length in cm
A= O. D(optical density)
By rearranging this equation you can calculate enzyme or substrate concentration.
Finally this will give you molar ratio.
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