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Getting Good Stuff from Wasted Stuff

The Cole-Fort Research Group

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Want to Avoid the Flu?

SoutheastAsianCooking

Shikimic acid

Tamifluâ

An H1N1inhibitor

Star Anise

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Shikimic Acid is Made by All Plants and Some Bacteria

More: http://chemistry.umeche.maine.edu/CHY431/Shikimate.html

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Well, trees are plants……

Pinus strobus (White Pine) Pinus strobus in front of Aubert Hall

…. And Maine has a bunch of ‘em

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So we went needle collecting in the University Forest north of campus

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We made conifer needle tea, and analyzed it

Soxhlet extractor

Agilent GC-MS System

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Some of Our Analytical Results

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Conifers Compared to Other Sources of Shikimate

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OK, we know it’s in there, now let’s get it out!

Step one:Chromatograph on ion exchange resin

Step two: Chromatograph on special silica gel

Step three: recrystallize

~ 3% isolated yield

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Now, scale it up

RotatingDigester

Giant tea bag

Filter

Chromatograph

Will it workon kg scale?

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Want to Live to Be 140?

Start with calorie restriction (CR): Calorie restriction means a low calorie diet containing all necessary nutrients CR extends the life of C. elegans, Drosophila, mice, and some primates Good health is maintained

Resveratrol Found in red wine (ng/mL) Claimed to be responsible for the “French paradox” Mimics the effect of calorie restriction

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OK, you’re a fat mouse, and we feed you resveratrol:

Insulin sensitivity increases Insulin-like growth factor (IGF-1) decreases Peroxisome proliferator increases Motor function improves Alzheimer’s peptide decreases

Result:

No type II diabetes Reduced risk of cancer Lower blood lipids You make the mouse Olympics

More: http://chemistry.umeche.maine.edu/CHY431/Resveratrol.html

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Resveratrol is Made by Many Plants

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So, we went out in the woods again, and cut down a bunch of trees, stripped the bark, and extracted it with acetone/water

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Why the bark?

Conifers store antioxidants in the bark to guard against fungi and bark beetles

Fungi use oxidative enzymes (laccase, peroxidase) to degrade bark; beetles attack partially degraded bark

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Source Resveratrol, mg/gNorway spruce 567Black Spruce 439White spruce 382

Eastern White Pine 7.1Japanese knotweed 523

Grape skins 32.5Low bush blueberries 12.8

Heirloom tomatoes 1.8Unsweetened chocolate 1.24

Quantitation by HPLC-MS, GC-MS, SIM

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Tons of bark are burned by mills every day!

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Shikimic Acid Resveratrol

Nilmini Vithanage (PhD) Regan LeBlanc (MS)

Alex Schaffer (REU) Melody Rhine (REU)Mark Rowe (OHS) Dustin Neidt (BS)Jess Richards (OHS) Sorel Edes (OHS)Zach Caron (OHS)Sorel Edes (OHS)Chi TruongJoe Dumont

Francis and Robin Avery, Brian Perkins

$$ USDA, NSF-EPSCoR, FBRI

Prof. Barbara Cole Prof. Joe Genco Prof. Ray Fort

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Extra:How doesResveratrolWork?

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Extra: How Does Resveratrol Work?