I am working on an enzyme assay for a fungal pectinase.I assayed the enzyme in different buffers from pH 1-12.5 However,the enzyme has good activities starting from pH1-10.5.
Is it possible to have enzyme activity over such a broad range of pH?
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$\begingroup$ Can you tell us which pectinase you are using? Besides that, is the activity the same over this pH range or does it vary? Can you show a curve of the activity plotted against the pH? $\endgroup$– Chris ♦Sep 2 '14 at 8:27
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$\begingroup$ Could you also give more details of methodology and controls - pectin can be hydrolysed non-enzymatically at low and high pH. $\endgroup$– Alan BoydSep 2 '14 at 8:31
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I actually doubt that the pectinase has such a broad pH range in which it works optimally. Searching the web I found two figures which support my doubts:
The first is from an article ("Immobilization of pectinase by adsorption on an alginate-coated chitin support") which compares the activity of native and immobilized pectinase under different conditions. The second is from the datasheet of a company which sells the enzyme. Both show an activity over 50% roughly between pH 3.5 and 5.5 but not at higher or lower pH values.
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$\begingroup$ I extracted the enzyme from 48 hours actinomycete culture ( submerged fermentation) the methodology I used is miller DNSA method $\endgroup$– OliSep 2 '14 at 8:58