Enzyme extraction/isolation - (Oct/15/2008 )

Hi there,
I am trying to extract enzyme from fungi..(such as xylanase, chitinase and beta galactosidase). But I didnt get a proper method. Can anybody give me some suggestion? and i would like to know, using HPLC or FPLC is suitable for enzyme study? I need to isolate the enzyme. Thanks in advance..

Abby05.

I just had a small look in a book I have. I am carrying out a similar thing for my project in college but I'm using a plant source. According to the book all you have to use is a pestle and mortar but freeze it in liquid nitrogen first. Then centrifuge.

Hi..
Thanks for your reply. But I am using liquid media and looking for extracellular enzyme. So I cant use pestle and mortar. I am still waiting for a proper method to extract enzyme. Crude extract is a mixture of all the enzyme and other chemical compounds, right? how to prepare this crude extract?

Let say I want to extract xylanase from fungi. But I do not know whether that spesific species of fungus has xylanase activity. Is it better to get the crude extract first? or do xylanase assay?

Thanks a bunch in advance..