Our team’s goal was to create a new method for chlorophyll removal from green seed canola oil. After months of hard work and experimentation, our team was able to demonstrate the following:

1. Our water soluble chlorophyll-binding protein, 6GIX, can be produced and purified
2. 6GIX can be emulsified and can be used to remove chlorophyll from canola oil

Using an IPTG-inducible system, our team was able to successfully produce and purify 6GIX in E.coli BL21(DE3). An SDS-PAGE gel with whole cell lysate and purified elutions of 6GIX without a signal peptide showed strong bands of the correct size (21kDa) in the whole cell lysate and elution 2. No bands were seen in the corresponding empty vector control lanes.

Figure 5: SDS-PAGE gel showing whole cell lysate and elutions after purification of no signal peptide 6GIX and empty vector control. Arrow denotes correct band length for 6GIX protein.

To make sure that the bands seen on the SDS-PAGE are of the correct protein, we ran a western blot using Anti-His MAb (from mouse) and Anti-mouse IgG conjugated with HRP antibodies. This western blot confirmed the identity of our protein at the correct band length.

Figure 5: Western blot showing whole cell lysate and elutions after purification of no signal peptide 6GIX and empty vector control. Arrow denotes correct band length for 6GIX protein.

We proved that our emulsification system successfully reduces the amount of chlorophyll in canola oil. with this THICC experiment