Submitted to: Applied Biochemistry and Biotechnology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 10/22/2008
Publication Date: 5/1/2009
Citation: Lee, C.C., Wagschal, K.C., Kibblewhite, R.E., Orts, W.J., Robertson, G.H., Wong, D. 2009. An alpha-glucuronidase enzyme activity assay adaptable for solid phase screening. Applied Biochemistry and Biotechnology. 155(1-3):314-320.
Interpretive Summary: Xylan polymer is the most common component of hemicellulose and represents a large and important renewable resource. In order to harness the full potential of xylan, the polymer must be enzymatically hydrolyzed to simple sugars. Glucuronic acid chemically modifies the xylan and prevents the complete hydrolysis of this polymer. Glucuronidase enzymes will remove the glucuronic acid from zylan and relieve the inhibition. We have developed an enzyme assay that allows for rapid discovery of new glucuronidase enzymes.
Technical Abstract: Glucuronic acid is a common chemical moiety that decorates the xylan polymer of hemicellulose. This chemical substituent impairs both enzymatic and acidic hydrolysis of xylosidic bonds. the fÑ-glucuronidase enzyme hydrolyzes the 1,2-linked glucuronic acid from the terminal, non-reducing xylose of xylo-oligosaccharides. There are relatively few fÑ-glucuronidase genes in the public databases. We have developed an assay with commercially-available reagents that can be used to search DNA libraries for fÑ-glucuronidase genes in a high throughput, solid phase activity screen.