Team:Rice University/RESULTS

A. Genetic Engineering and Part Construction

A detailed summary of parts, design considerations, and genetic engineering progress can be accessed at the CONSTRUCTS page.

Please visit our Notebook for a summary of labwork and protocols.

B. Yeast Transformation

We are currently accumulating data for plasmid transformations into our brewing strain, SAB-Hefe. Data will be updated soon.

C. HPLC Data

To analyze our beer samples for resveratrol content, be will be using High Performance Liquid Chromatography (HPLC), which will allow us to separate the metabolites produced by the yeast and analyze these compounds by spectrophotometry. By comparing HPLC chromatogram peaks of metabolites produced by our yeast with a resveratrol-only standard, we can identify if resveratrol is being produced, and at what quantities. We will be monitoring the production of resveratrol and the consumption of p-coumaric acid, which will only be added to the media for the 4CL::STS-integrated yeast; the 4CL::STS+TAL . Below, we show our initial data for HPLC calibration curves using known quantities of resveratrol and p-coumaric acid standards and test chromatograms using extracts from different wine samples.

Figure C1. Example HPLC chromatogram of a trans-resveratrol standard.

HPLC: Calibration

Figure C2: trans-resveratrol standards (Sigma). Different serial dilutions are shown: 20 ug/mL (red), 10ug/mL (purple), 5ug/mL (blue), 2.5ug/mL (green).

Figure C3: p-Coumaric acid standards (Sigma). Different serial dilutions are shown: 18.5ug/mL (red), 9.25ug/mL (purple), 4.63ug/mL (blue), 2.3ug/mL (green).

Figure C4: trans-resveratrol and p-coumaric acid separate at different times; this will allow us to resolve both molecules in a complex mixture, such as a yeast extract. Shown is 18.5ug/mL of p-coumaric acid and 20ug/mL trans-resveratrol of

HPLC: Wine Extract Tests

HPLC: Fermenation batches

Fermentation

Coming Soon. For a sneak preview, check out the Gallery