I'm currently extracting GAD enzyme (Glutamate decarboxylase) by doing enzymatic lysis onto the fungus Aspergillus Oryzae.The lytic enzymatic cocktail was Yatalase, which composing of Chitinase, Chitobiase, Chitosanase, beta-1,3-glucanase and Protease.
I used the cocktail (10 mg/ml) to lyse 5, 10, 20, 30, 40, 50, & 60% w/v of fungal concentration. After incubation time (60 mins), the supernatant was collected for determining total protein (Bradford reagent) and enzymatic activity (GABA Assay).
Here's my finding: total protein increased proportionately with fungal cell concentration. However, total activity and specific activity of GAD enzyme decreased from 30% w/v fungal cell concentration. IMO, I thought total activity of GAD would increase with total protein i.e. more proteins released from fungal cells, more GAD collected as well. Is this result usual or unusual? What phenonmenon caused it?
Fungus (5, 10, 20, 30, 40, 50, & 60% w/v) Undergo enzymatic lysis. Enzyme of interest: GAD
Total protein: increases
Total activity of GAD: decrease
Specific activity of GAD: decrease
Not sure how to interpret the result.
Edited by schroddycat, 02 September 2012 - 12:48 AM.