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| Content Provider | Springer Nature Link |
|---|---|
| Author | Muir, Ryann M. Ibáñez, Ana M. Uratsu, Sandra L. Ingham, Elizabeth S. Leslie, Charles A. McGranahan, Gale H. Batra, Neelu Goyal, Sham Joseph, Jorly Jemmis, Eluvathingal D. Dandekar, Abhaya M. |
| Copyright Year | 2011 |
| Abstract | Gallic acid (GA), a key intermediate in the synthesis of plant hydrolysable tannins, is also a primary anti-inflammatory, cardio-protective agent found in wine, tea, and cocoa. In this publication, we reveal the identity of a gene and encoded protein essential for GA synthesis. Although it has long been recognized that plants, bacteria, and fungi synthesize and accumulate GA, the pathway leading to its synthesis was largely unknown. Here we provide evidence that shikimate dehydrogenase (SDH), a shikimate pathway enzyme essential for aromatic amino acid synthesis, is also required for GA production. Escherichia coli (E. coli) aroE mutants lacking a functional SDH can be complemented with the plant enzyme such that they grew on media lacking aromatic amino acids and produced GA in vitro. Transgenic Nicotiana tabacum lines expressing a Juglans regia SDH exhibited a 500% increase in GA accumulation. The J. regia and E. coli SDH was purified via overexpression in E. coli and used to measure substrate and cofactor kinetics, following reduction of NADP$^{+}$ to NADPH. Reversed-phase liquid chromatography coupled to electrospray mass spectrometry (RP-LC/ESI–MS) was used to quantify and validate GA production through dehydrogenation of 3-dehydroshikimate (3-DHS) by purified E. coli and J. regia SDH when shikimic acid (SA) or 3-DHS were used as substrates and NADP$^{+}$ as cofactor. Finally, we show that purified E. coli and J. regia SDH produced GA in vitro. |
| Starting Page | 555 |
| Ending Page | 565 |
| Page Count | 11 |
| File Format | |
| ISSN | 01674412 |
| Journal | Plant Molecular Biology |
| Volume Number | 75 |
| Issue Number | 6 |
| e-ISSN | 15735028 |
| Language | English |
| Publisher | Springer Netherlands |
| Publisher Date | 2011-01-30 |
| Publisher Place | Dordrecht |
| Access Restriction | One Nation One Subscription (ONOS) |
| Subject Keyword | AroE Gallic acid Juglans regia Escherichia coli Shikimate dehydrogenase Walnut Tannins Plant Sciences Plant Pathology Biochemistry |
| Content Type | Text |
| Resource Type | Article |
| Subject | Genetics Plant Science Medicine Agronomy and Crop Science |
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