Date of Award
2010
Degree Type
Thesis
Degree Name
Master of Science
Program
Biology
Supervisor
Dr. Mark Bernards
Second Advisor
Dr. Susanne Kohalmi
Third Advisor
Dr. André Lachance
Abstract
The conversion of p-coumaroyltyramine to caffeoyltyramine catalyzed by p-coumaroyltyramine 3-hydroxylase (CYP98A) appears to be specific to suberization and could serve as a marker for wound-inducible suberization. The expression of this enzyme could be used as a tool for future studies of suberin biosynthesis. Five potential candidate CYP98A genes from Solanum tuberosum EST databases were determined (designated pCT3Hl through pCT3H5). The pCT3H2 gene was sequenced, cloned into the pTrcHis2B expression vector and transformed into TOPlO E. coli. pCT3H2 expression was induced and the recombinant protein was tested for the catalytic conversion of p- coumaric acid, p-coumaroyltyramine, p-coumaroylshikimate, and p-coumaroylquinate using HPLC. Although expression was successful, catalytic activity was not demonstrated. It is possible that post-translational modifications, such as insertion of a heme necessary for functionality, are not properly carried out in E. coli. Further studies are necessary to determine whether pCT3H2 acts as a CYP98A in Solanum tuberosum
Recommended Citation
Anderson, Stephanie, "FUNCTIONAL CHARACTERIZATION OF A PUTATIVE WOUND-INDUCED p-COUMARO YLTYRAMINE 3-HYDROXYLASE FROM SOLANUM TUBEROSUM" (2010). Digitized Theses. 4480.
https://ir.lib.uwo.ca/digitizedtheses/4480