Secondary metabolites are often restricted in their distribution to different groups of organisms. For this reason, attempts to study these often useful and interesting products of metabolism require an ability to work in a diversity of non-model species. Methods for gene discovery with low investment and high efficiency are needed to effectively identify the biosynthetic genes in these diverse pathways. During this work, a workflow for efficiently identifying biosynthetic genes was developed and applied to Amaryllidaceae alkaloid biosynthesis. Genes discovered during this work include a norbelladine 4’- O-methyltransferase, a cytochrome P450 capable of phenol-phenol coupling 4’-O-methylnorbelladine to noroxomaritidine, and a short-chain dehydrogenase/reductase capable of forming norbelladine from tyramine and 3,4-dihydroxybenzaldehyde. These enzymatic discoveries support the future application of this workflow to other biosynthetic pathways and organisms.
|Advisor:||Kutchan, Toni M.|
|Commitee:||Haswell, Elizabeth, Henderson, Jeffrey, Jez, Joseph M., Kunkel, Barbara, Mockler, Todd|
|School:||Washington University in St. Louis|
|Department:||Biology and Biomedical Sciences (Plant and Microbial Biosciences)|
|School Location:||United States -- Missouri|
|Source:||DAI-B 77/05(E), Dissertation Abstracts International|
|Subjects:||Plant sciences, Biochemistry|
|Keywords:||Amaryllidaceae, Amaryllidaceae alkaloids, Cytochrome P450, Galanthus, Narcissus, Norbelladine|
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