Dear researchers, I need to concentrate a fungal cellulases crude extract, even if I have tried a column concentration device, I lose enzyme activity, so is there an alternative way? Thanks, Veronica
Pass your crude extract thorugh at 10KD/30KD TFF membrane if your volumes are large or use 10KD/30KD centrifugal filter devices in case your volumes are low to concentrate cellulase enzyme. It should be kept in mind that the membrane should be made of Polyether sulfone (PES) rather than cellulose acetate. Good luck for your research.
Precipitate enzyme with ammonium sulphate at 70% saturation from crude extract solution at pH 4.5-4.8 with almost 100% yield.
As Deepti Agarwal rightly said that you can pass enzyme through UF of polymethyl sulphonate to concentrate it 8-10 times as cellulases are sturdy enzymes. Also you can try ammonium sulphate precipitation.
Thanks a lot, anyway as by PES membrane ultrafiltration devices I loose activity, I was just looking for a different way of acting: I will surely try precipitation!
If you want to go for precipiation, I will suggest you to go for chilled acetone (80%) precipitation along with AmSO4 as suggested y Vladimir. Wish you good luck.
Can you tell me what is the specific activity of your crude enzyme? If you have less amount of protein (enzyme) then in your precipitation method (amm. sulfate ppt.) you will not get any good result. So it is my suggestion to get more protein (mg) in crude sample by optimizing the conditions. Hope you will be benefited. All d best..
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