Vitamin Experiment
After we found that UV has an impact on Vitamin, we than wanted to look further inside the relationship between UV and Vitamin. We conducted an experiment on different kinds of bottles and the degradation rate of Vitamin under sunlight. While we visit VEDAN, they told us that most of the refreshing drinks contain lots of Vitamin (especislly Vitamin B) which are easily deteriorated by UV. Generally, manufacturers put their drinks inside brown bottles to protect their product from UV. Than we sent our samples to Food Industry Research and Development Institute(Taiwan) to examine the amount of Vitamin B.
However, we also found out that the percentage of Vitamin B remain dropped off rapidly to 30% and 14% in brown bottle and transparent bottle respectively. This result simply means that those refreshing drink need to be protect from UV, and this is time our sticker can show its capabilities.
Experiment
Our product aims to reach a high quality of “long-term monitor”, which means considerably stable expression of color is needed. Thus, we choose chromoprotein as our reporter protein due to its steadily expression and low half-life degradation rate. (See more in here)
cj-Blue
We add a LVA tag to cjBlue in order to accelerate its degrade speed. (See more in here)
Concentration simulation of cjBlue
Chromoprotein
According to our model of chromoprotein
Expression
Temperature
35ﾟC / 37ﾟC / 39ﾟC / 32ﾟC
Chromoprotein o.d. value
(1) RFP (E1010): 607nm
(2) BFP (k592009):588nm
(3) cjBlue (k592011):610nm
Culture Setting
(1) 32ﾟC, 200rpm, culture12hr (15ml) *continue to log phase
(2) Add (15ml) LB *re-culture
(3) TeﾟC, continue cultural
(4) Take samples from the centrifuge tube per hour, and narrow the time range and improve the accuracy.
UV
Culture setting
(1) 37ﾟC, 200rpm, culture 12hr (15ml) *continue to log phase
(2) Add (15ml) LB *re-culture
(3) Re-culture at 37ﾟC under UV
(4) Take samples from the centrifuge tube per hour, and narrow the time range and improve the accuracy.
Expression measurement setting
(1) Extract: Take (3ml) bacteria liquid, and centrifuge at 14000rpm, 1min
(2) Add 1ml PBS, pipetting+vortex
(3) Measure the O.D. value
Results
This is the result of the biobrick K2383200 and K2383302. We culture the bacteria for 15 hours. The concentration in the figure is the concentration the bacteria. 100%= the origin liquid. We do dilution to simulate the pass of time. The minimum amount for naked eyes to recognize is 50%.
