SDS Page:
After DNA fragments were assembled and ligated, DNA gel analysis was used to estimate the size. Additionally, the compatibility of the size and protein expression was confirmed by the SDS-PAGE analysis.
The results of the SDS-PAGE study
performed on our engineered E. coli are shown in the attached figure.
![](https://static.igem.wiki/teams/4689/wiki/results-page/res-1.jpg)
The study illustrates the production of prk, rbCS, rbCX, and rbcL at molecular weights of 35 kDa, 10 kDa, 14.4 kDa, and 48 kDa, respectively. As it is the case of the dual plasmid, the production of kivD and adhA were also confirmed separately, with molecular weights of 61 kDa and 35 kDa, respectively.
![](https://static.igem.wiki/teams/4689/wiki/results-page/res-2.jpg)
[1]TIME VS XYLOSE CONSUMPTION
A growth curve was monitored for Engineered E.coli K12 containing prk and RuBisCo, showing a rapid and increasing uptake of xylose over time.In engineered E.coli, there are two primary sources for CO 2 metabolism, namely xylose and CO2 itself. Our experiments were carried out using approximately under 5% CO2 and 1% xylose.
![](https://static.igem.wiki/teams/4689/wiki/results-page/xylose-consumption-engineered-e-coli-k12-prk-rubisco.jpg)
![](https://static.igem.wiki/teams/4689/wiki/results-page/xylose-consumption-e-coli-k12-control.jpg)
![](https://static.igem.wiki/teams/4689/wiki/results-page/average-xylose-consumption.jpg)
[2] TIME VS RUBISCO+PRK
Batch fermentation profile of E. coli K12 engineered strain containing prk and RuBisCo was conducted.Bacterial strain was cultivated in M9 minimal media with xylose in a bioreactor to favor growth of recombinant bacteria supplemented with 1% Xylose and supplied with 5% CO2 at 37°C under pH control.The optimal pyruvate production was observed in the engineered bacteria which is confirmed using the Methyl red test .
![](https://static.igem.wiki/teams/4689/wiki/results-page/time-vs-od-at-600nm-engineered-e-coli-k12-prk-rubisco.jpg)
![](https://static.igem.wiki/teams/4689/wiki/results-page/negative-control-e-coli-k12.jpg)
![](https://static.igem.wiki/teams/4689/wiki/results-page/growth-curve-engineered-k12-prk-rubisco.jpg)
[3] TIME VS ISOBUTANOL PRODUCTION
Batch fermentation profile of E. coli K12 engineered strain containing prk,RuBisCo,kivD and adhA.Bacterial strain was cultivated in M9 minimal media with xylose in a bioreactor to favor growth of recombinant bacteria supplemented with 1% Xylose and supplied with 5% CO2 at 37°C under pH control.Isobutanol is an extracellular product confirmed by the Lucas test to check for it’s presence at the end of the engineered cycle.
![](https://static.igem.wiki/teams/4689/wiki/results-page/time-vs-isobutanol.jpg)