Team:Evry/Medals

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Let's PLAy project - Bioproduction of PLA

Medals

Bronze

Status: Awarded


Criteria Fulfillment Links
Register and attend

We are registered! After having a hard-working but amazing summer, most of us went to the Giant Jamboree to proudly present our work!

Deliverables

Wiki: Here it is, enjoy it!

Poster: We presented our poster in the Giant Jamboree.

Presentation: We defended our project in the Giant Jamboree.

Project Attribution: Check out our wiki!

Registry Part Pages: Yes, linked on our wiki.

Sample Submission: Our beloved team owl brought them to the USA.

Safety forms: We indeed received safety formation, and we studied safety issues involved in our project.

Judging Form: Links activated, 150 words for Prizes written poetically.

Attribution

We described both team work and external help we had.

Part / Contribution

A Pseudmonas putida promoter inducible with Cyclohexanone, studied for the expression of the heterologous genes needed for PLA production.

Silver

Status: Not Awarded


Criteria Fulfillment Links
Validated Part / Validated Contribution

A Pseudmonas putida promoter inducible with IPTG. Potentially used for the expression of the heterologous genes needed for PLA production. It has been characterized with GFP as reporter of expression, and results have been contrasted and validated with literature.

Collaboration

We performed experiments for both Paris Bettencourt and Imperial College teams. We organized the European Experience with IONIS team and we did several smaller collaborations.

Human Practices

We started by a survey on the consumption of bioplastics, and ended up with a MOOC. Check the intermediate processes on our wiki!

Gold

Status: Not Awarded


Criteria Fulfillment Links
Integrated Human Practices

The issues investigated led to think about designing a practical way to implement PLA. We decided to design a DIY bioreactor that would demonstrate PLA production to be not expensive and would, at the same time, optimize it. Besides, we conceived and did a MOOC as consequence of awareness study. Check on the wiki!

Improve a previous part or project

Yale 2013 had previously worked on PLA, but using E.coli. We corrected an error on their original BioBrick characterization and we improved BioBricks of PhaC and Pct by optimizing them to P.putida.

Proof of concept -
Demonstrate your work -


Prizes

Status: Not Awarded


Prizes Description Links
Integrated Human Practices

After conducting a survey on our project, it appeared that people would prefer resorting to bioplastics in their daily life rather than petroleum-based equivalents. Yet the consumption of bioplastics remains lower. Therefore, we decided to focus on solutions to tackle this paradox. The first step was to carry out a pedagogic communicative approach to arise interest on synthetic biology, and then on PLA. That is why we decided to focus on MOOCs as an effective tool to spread knowledge on this topic. Besides, we noted that globally, public policies tend to gradually ban plastic consumptions and that could be related to restrictive legislations directed at petroleum-based plastics. Thus, we worked on describing an efficient DIY bioreactor that would push PLA scale-up in a cheap manner. Additionally, facing the controversy on PLA degradation, we have highlighted the role of skepticism on science and performed a simple experiment of plastics degradations.

Education and Public Engagement

Producing a bioplastic involved a broad range of topics from microbiology to genetic engineering and we asked ourselves how we could efficiently present them to a wide audience. At first, we met different kinds of publics from high schools to universities, through presentations and fairs. Discussions were interesting, but the impact was relative. Secondly, we looked for a tool enabling interactions with a large number of people on a given topic, on a short period of time. The MOOC concept seemed to fill those criteria. As we agreed on appraisals made on MOOCs, we decided to focus on its disadvantages to counterbalance our initial biaised opinion. That’s why, we experimented MOOCs format with high school and University students, in order to have external points of view. We also created a survey for gathering opinions and spread information; translated Wikipedia for iGEM competition; and participated in public events.

Model

In silico experiments can help us improve the basic design of our system. In our case, we have focused in two strategies: Flux Balance Analysis and a system of Dynamic Regulation. For FBA, we have modified a published SBML model of P.putida KT2440 to include our PLA production reactions and we have studied the tuning of cellular metabolism through genetic engineering, enzyme engineering and biochemical optimization of growth conditions. The results have been taken into account in wet-lab experiments of best carbon source selection. For Dynamic Regulation, we have designed a genetic-metabolic circuit regulating the pathway through feedback loops. Using biosensors and repressible promoters, the cell can find its own equilibrium to make PLA. We have studied it from two different approaches: rule-based modeling and ODEs in genetic circuitry. We would integrate the dynamic design in the whole system of optimization and improvement of the initial implementation.

Applied Design

Pseudomonas putida was elected as chassis for PLA bioproduction because its ability to produce polymers, conferring advantage on polymerization bottlenecks. Inducible promoters were used in order to control cell growth and production rates. An extra gene for increasing precursor production was implemented. Optimal growth conditions were investigated through Flux Balance Analysis, by testing optimal carbon source. We conceived and developed in silico a dynamic regulation system based on biosensors and inducible promoters to optimize biosynthesis by means of transcription factors binding precursors from the pathway to assess the optimization in a design-build-and-test iterative fashion. Bioprocess parameters based on experimental cell growth data were described. We designed and built a DIY bioreactor and described its optimal running. A DIY extrusion system and a roller were built for further downstream processing of the biopolymer to provide the whole system with an affordable manner to obtain and prepare the final product.