Cells are disrupted and debris are removed using a 0.2 μm filter to obtain enzymes. However, after filtering(remove debris) , the value of the titre is very low. why happen to low enzymes titre?
Step : culture => disruption(high titre value) ==> filter(remove debris) => titre(Low titre value)
The enzyme may be sticking to the filter and the filtration equipment. Instead of filtering to remove debris, consider centrifugation.
The enzyme may be unstable in the extract, for example because of proteolysis, and loses activity over time because of that, rather than because of filtration.
Assuming the enzyme is not sticking to filter material and the case was encountered after applying centrifugation similarly...What is the cell lysis buffer composition?...
If the purposed enzyme is not secreted it should exist either membrane-bound (anchored)-non-specifically interacted-bound to the debris or exist as aggregated (inclusion bodies) in the cytosol. During filter or centrifugation application you may lose targets due to the abovementioned reasons. At this point, lysis buffer composition with the preferred physical treatment option is critical to assess your loss...
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