I am working with immobilized enzymes i want to calculate immobilization efficiency
Immobilized enzyme activity and Immobilization efficiency are two different things.
Immobilization efficiency is the protein binding capacity of a particular matrix/support that you are using. The percent of the total protein bound onto the matrix indicate its immobilization efficiency.
Activity of immobilized enzyme can be calculated as U/mg of your immobilized biocatalyst.
Good luck
Also if you want to calculate the binding efficiency
; You can calculate the total protein of stock solution before immobilization,
After immobilization you can separate the supernantent and protein bound matrix then you have to calculate the amount of protein [unbound protein] left in the supernatent. Now you can calculate the ampount of protein bound onto your matrix by subtracting the protein of supernatent from total protein. You can calculate the percentage of same.
Also during washing of your matrix you may loose some loosely bond or unbound prtein from your biocatalyst.
Thank you for your valuable suggestions Sir. I will try to implement it.
The observed of immobilized enzymes is not calculated; it is empirically determined using a standard activity assay. It corresponds to the expressed activity of the catalyst. Once you have already determined this observed activity you must divide it by the total activity you added to immobilize. There you are the immobilization efficiency. Take a look to the attached paper.
Good luck!
Also remember when measuring the activity of the immobilized enzyme that its apparent response to, for example, pH may be different depending upon the immobilization matrix. You immobilized enzyme may have an apparent (or real for that matter) pH optimum above or below the soluble enzyme: apparent Km can also change so you need to confirm that the conditions you are using to assay the enzyme are optimum in each case before comparing immobilized to soluble activity.
Thank you so much for your valuable suggestions, I will implement your suggestions
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