I'm currently conducting research on the production of xylanase, pectinase and protease via solid state fermentation. The crude enzyme I harvested from each set of SSF is my primary source of each enzyme I mentioned earlier.
From my literature review it is okay to use sodium acetate buffer for the extraction of xylanase and pectinase. How about protease? does sodium acetate buffer suitable for the leaching out of protease from the SSF?
Since most of the pectinase work in acidic pH therefore the extraction buffer used is generally citrate or acetate. Protease can be neutral, acidic or alkaline. Based on type of protease secreted by your fungi during SSF, you need to choose the buffer system. When I was doing my Ph.D. even physiological saline or 1% Tween-80/20 used to be sufficient for leaching out protease from the SSF...
I think that sodium acetate buffer is very acid for proteases. You can use a neutral buffer, for example, sodium phosphate at pH 7, or you can use distilled water until you know your enzyme.
The reasonable and general way is the use of the buffer with the pH at which the enzyme has optimum /maximum activity to keep the enzyme on it's suitable conformational structure gives maximum activity. In addition, the buffer concentration should be at gentle values such as in the range of 0,03- 0,5 M depending on the enzyme type. You can determine the suitable buffer kind, it's pH vand also concentration values from the related literature on the base of your specific enzyme.
I didn't add any additional chemical on my substrate prior to fermentation.
The choice of buffer depends upon the optimal pH of the enzyme to be extracted, which again lies with the source of the enzyme. Of-course you can use sodium acetate buffer if it is acidic pH range.
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