Team:TU Delft/Project/conclusions

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Conclusions

Our goal was to tackle the biological conversion of hydrocarbons in an aqueous environment. The basis of the project was generating a "biological chassis", which provides the framework for varying and multiple characteristics needed for the conversion of hydrocarbons, including considerations like conversion ability, hydrocarbon tolerance/solubility and halo (salt) tolerance. This chassis could than be used in for example specifically the biological degradation of oil particles in oil sands tailing water.

Alkane Degradation

Survival

Salt tolerance

Our biobrick has enabled us to increase the salt tolerance of E.coli by an average of 20%. But due to the range of effects caused by increased salt stress, complete tolerance using a single protein is impossible. As such we hope to have made a first step and that the future iGEM teams will be able to build upon this knowledge.

Solubility

To overcome the mass‐transfer limitation between the water and oil fase, a gene encoding for AlnA, a protein with emulsifying properties was expressed. The increased solubility of about 20% was determined by a new method. We suggest that in future research the protein is tagged, so it can be isolated with higher purity.

Sensing

Anderson RBS Characterizations

In order to tune the protein expressions of the alkane degrading genes we've characterized 5 members of the Anderson RBS family using an improved protein production model taking dilution into account. The following relative efficiencies were found:

RBS Efficiency
J61100 0.03
J61101 0.12
J61107 0.08
J61117 0.02
J61127 0.07
B0032 0.30