Xylanase Enzyme by Andru O’Farrill, Dr. Ray Fort & Dr. Barbara Cole
The work conducted will consist of two interconnected types of research based on the same topic. The research will include both chemistry lab work, as well as in the chemistry computer lab classroom. The experiments will be done with the enzyme, xylanase, and the substrate xylobiose. Xylobiose is a product that is formed from the hemicelluloses in trees. Currently there is research being conducted in the chemistry lab to use the xylanase enzyme to cleave hemicelluloses into desired lengths. One of the molecules that should theoretically be detected is the xylobiose molecule. Using the xylanase enzyme the linkage between two xylose units in xylobiose should be cleaved. There will be two aspects of this that will be studied.
First, a way to identify the xylobiose molecule must be found using gas chromatography/ mass-spectrometry (gc/ms), and liquid chromatography/ mass-spectrometry (lc/ms), most likely lc-ms. These trials should theoretically find a way to determine if xylobiose is present in a mixture, and to see if the particular strand of xylanase being used is an efficient enzyme. Secondly, the research will consist of altering the reaction that is obtained when xylan is hydrolyzed by xylanase enzymes. Temperature and time will be altered to find the optimum conditions to drive the reaction.
In the computer lab, the computer simulation program Autodock will be used to examine how the xylobiose binds to the enzyme. By setting up a 3-D grid across the enzyme, and analyzing the interactions at different points on the enzyme, the location on the enzyme where xylobiose binds, and the best conformation of the xylobiose for this reaction to occur can be found. Additionally, different conformations will be analyzed to see the effects on the enzyme’s ability to catalyze the reaction. This will in turn give a good idea of what must be done in the lab to accomplish the selective cleavage of the hemicellulose bonds., and will reduce the amount of different xylanases that will have to be bought for lab experiments, and lab work done. This research will also help understand the same problems that are being found under the larger scale hemicellulose molecules.
Interview with Andru July 5, 2007
