An experiment to measure the optimum ph of an enzyme

Question by collette512bbi Reward: 25 Pond Points + 10 Knowledge Tokens Status: Pending a resolution
I need an experiment to show me how to measure the optimum ph of an enzyme please. I chose an experiment with photographic film placed in acid, alkili and enzyme solution. So really I need to find the effects of a protease on gelatin at different ph's.



Reply from nicomarm User Rating:  1380 Knowledge Tokens
One way to do it will be incubation of the protease with the substrate 0.2% gelatine at 30C for 10 min. use different pH 6.0,6.5 7.0 7.5 8.0 8.5 9.0 or other ranges if you have some information about the particular enzyme. I would run samples on electrophoresis gels and staine with coomassie blue. Run a standard gelatine in one line .Measurement of the lower intensity band for gelatine will show the highest enzyme activity . Give me more details about the photographic film and I will try to adjust the assay to it.
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