Last week I spent a lot of time doing research on the celullase complex and the structure of cellulose itself. Mat and I are still trying to come up with a way to determine whether the cell wall has been degraded and to what extent by the cellulase solution before starting the extractions. Initially, we thought we may use the Maltose color reagent and spectrophotometry to generate a curve to find the optimal pH and Temperature within a range that won't denature DNA molecules. So far we have the optimal pH of the complex at between 4-5 and temp at 60-70 C, but the low pH may cause severe damage to DNA. So far I think we are going to keep the pH at 5 for the digestion time and then proceed with the buffer and the same protocol I have been using. I will be carrying out two extractions, one with chopped pieces of tissue and the other one with ground with mortar and pestle as usual.
These couple of days we get free I have been wondering whether this could work. Ideally it will be a very nice addition to the protocol, that is to have enzymes aid in the breakage of cellulose. However, as of today I have not found any information on any fungus attacking Palo Verde trees, except for one that grows in its roots. So all my planing may actually not even work. Either way, I have become very interested on enzymes and specially those in fungi. So although this may or may not work for this project, it may be the basis of my own project next semester.
The though Sonoran desert tree may not even be susceptible to enzymatic digestion.
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