We produced the crude enzyme solution from white-rot fungi grown on apple waste. We did not do any purification and therefore the compounds in apple wastes are present in the solution. The target enzymes are laccase and lignin peroxidase. according to standard methods for laccase, we performed the oxidation of ABTS. But we saw a very small activity. Now I am thinking about the effects of antioxidants and polyphenolic compounds that are present in solution. Does anybody have any idea?
In my opinion, may be the low activity shown in your lab related with the low amount quantity produced from pomace material. The low quantity yield of white-rot fungi may be inhibited by phenolic content in pomace itself, Thus the activity of laccase depend of the quantity of white-rot fungi produced, not phenolic content in pomace. For considering, In our laboratory we analyzed wild grown mushroom white-rot fungi from tropical plant showed moderate activity, and available for future isolation the enzyme. This enzyme prepare for green synthesis modification of phenolic compounds to enhance the bioactivity.
Simply change the method for laccase activity. Use siringaldazine as a substrat.
Hi Mehrdad Taheran
Body can deactivate free radical different ways so antioxidant can measure different way. You can check antioxidant through FRAP (Ferric reducing antioxidant power) method. Please see the attachment it may help you.
Try for the peroxidase (Ascorbate peroxidase, glutathion peroxidase and guicol peroxidase ). it is the maximum in pomacea so either from the enzyme assay or native page you will get some result.
Pomace is rich in lot of polyphenols, for example quercetin is powerful and very common in apple.
Peroxidase (Ascorbate peroxidase, guicol peroxidase and glutathion peroxidase) are the common antioxidants present in pomacae. you can go for the assay and activity staining of the native gel for these enzymes.
There are many bioactive polyphenols within apple pomace, many of which display antioxidant activity. Some examples include catechins and their dimers (procyanidins), trimers etc, quercetin and quercetin glycosides among many others. There are several articles you can review some of which you can view from these links.
http://www.sciencedirect.com/science/article/pii/S0308814699001673
http://www.ncbi.nlm.nih.gov/pubmed/19203755
http://www.ncbi.nlm.nih.gov/pubmed/21077127
Dear Friends, Now I am sure that we have antioxidants in pomace. but is there any idea about their inhibitory effects on laccase?
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