Team:CSU Fort Collins/Design

COLORADO STATE
UNIVERSITY iGEM

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Design

Proposed Process

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Evaluating Design

We spoke to employees at the City of Fort Collins to discuss how our process would integrate into frying oil recycling in our community. In Fort Collins, most restaurants pay a company to retrieve and recycle their oil into biofuels. Our process could allow for restaurants to break even or even potentially be paid for recycling their spent frying oil. And while Colorado is very eco-friendly, many places in the United States and around the world could benefit more incentive them to upcycle their oil. Our use of frying oil waste as an alternative substrate for chemical production would not necessarily replace, but expand the amount of frying oil waste recycled instead of sent to landfills.

The production of cytokinins in bacterial hosts also offers an important alternative to the time- and cost-intensive process of extraction from plant tissues. Our proposed use of biological hosts to manufacture reagents is part of a larger movement in the synthetic biology community to work towards safer production. Ideally, products would be transported outside of the cells. This way, the current standard of using harsh chemicals to extract the product is unnecessary.

The cheap production of zeatin would also allow for its expanded use in industry and agriculture. There is the potential for this to provide an alternative to the current standard of pouring enormous volumes of herbicide and pesticides onto crops. Zeatin could also be used in areas with poor crop performance to increase yields.

Functional Prototype Creation

Our team attempted to build a functional prototype of both the trans-zeatin production and frying oil breakdown steps of our process.

We grew our strains with the trans-zeatin biosynthesis pathway in 1L bioreactors over 72 hours. While the results of this experiment were inconclusive, we were able to design an experiment which represents a step towards the scaling up of our process.

We were able to show a functional prototype of our frying oil digesting strain. We developed an experiment which grew our strain on frying oil waste (both at 100% and 50% concentrations) donated to us by a local restaurant,The Mayor of Old Town. The results of this experiment showed that our lac promoter:fadD:fadL construct improved the cells’ ability to grow on actual frying oil waste.