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Team:TU Delft/Project/conclusions
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===Survival=== | ===Survival=== | ||
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====Salt tolerance==== | ====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. | 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. | ||
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| + | ====Solvent tolerance==== | ||
| + | Our biobrick has enabled us to increase the solvent tolerance of ''E.coli'' when n-hexane is present in the culture medium at high concentrations. According to our findings, the growth rate of ''E.coli'' is improved 60% at a n-hexane concentration of 10%(v/v). | ||
===Solubility=== | ===Solubility=== | ||
Revision as of 12:33, 27 October 2010
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.
Solvent tolerance
Our biobrick has enabled us to increase the solvent tolerance of E.coli when n-hexane is present in the culture medium at high concentrations. According to our findings, the growth rate of E.coli is improved 60% at a n-hexane concentration of 10%(v/v).
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
After the pre cultures are grown in glucose, it needs to switch to alkanes as a feedstock. Therefore we designed a biobrick switch that can sense the absence of glucose: pCaif.
CONCLUSION
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 |
| [http://partsregistry.org/Part:BBa_J61100 J61100] | 1.20% |
| [http://partsregistry.org/Part:BBa_J61101 J61101] | 11.9% |
| [http://partsregistry.org/Part:BBa_J61107 J61107] | 7.70% |
| [http://partsregistry.org/Part:BBa_J61117 J61117] | 1.26% |
| [http://partsregistry.org/Part:BBa_J61127 J61127] | 6.52% |
| [http://partsregistry.org/Part:BBa_B0032 B0032] | 30.0% |
