Objective

We reached out to the de Lorenzo Lab to learn how to select optimal plasmids and promoters for Pseudomonas species capable of synthesizing PHB from TPA. Their advice helped us narrow down the SEVA database and support the idea of using a highly effective expression system, XYIS/PM. Their guidance was instrumental in helping us avoid potential experimental errors and in identifying the most suitable Pseudomonas chassis for our work.

Takeaways

The correspondence provided several key insights that helped guide our project. They first recommended Pseudomonas putida KT2440 as a reliable chassis, highlighting its efficient natural excretion system. They also suggested the XyIS/Pm system, known for its tight regulations and high induction capabilities. In addition, they shared the pSEVA228 plasmid (pSEVA228) as a valuable resource. Finally, they emphasized the importance of using a growth medium with a high carbon-to-nitrogen ratio to stress the cells and promote PHB accumulation.

Implementation

After our interview, we followed the recommendations that were provided. We transitioned from P. stutzeri to Pseudomonas putida KT2440 as our chassis. Next, we selected and designed an appropriate SEVA plasmid from the XyIS/Pm expression system. Once the parts are secured, we cloned our TPA-to-PHB pathway genes and transformed them into our host. For cultivation, we grew the engineered strain in a minimal M9 medium, designing it to provide TPA as a carbon source while limiting nitrogen availability. Finally, we induced protein expression and triggered the production of enzymes that will activate PHB synthesis. Therefore, their advice provided us with a clear pathway for our experimental approach.

BluePHA

Overview

We met with Dr. Chitong Rao, chief scientist at BluePHA Co., Ltd., a leader in synthetic biology known for their production of fully biodegradable bio-polymer polyhydroxyalkanoates (PHAs) to replace traditionally made plastic. Co-founded by two former iGEM team leaders at Tsinghua and Peking University in 2010, their expertise in advising businesses on reducing their dependence on conventional plastics served as an example we hoped to learn from. We discussed their work on implementing a circular economy and optimizing productivity in biological processes, and we delved into the key considerations for life cycle analysis and biosafety protocols. Throughout the correspondence we were given early access to unpublished data on the LCA.

Objectives

Our primary goal was to gain insight from BluePHA's expertise in sustainable biopolymer production, especially on experiment protocols and genetic constructs. Also, a goal was to gain potential insights in drafting a Life Cycle Analysis (LCA), especially on the quantitative calculation of determining the carbon footprint of PHA. PHA is a fully biodegradable family of polymers derived from biomass or waste. Since one of our team's goals is to enhance the production of poly(3-hydroxybutyrate) (PHB), a specific type of polyhydroxyalkanoate (PHA) that can be depolymerized into the high-value beta-hydroxybutyrate (BHB), understanding the optimization of this process became a key objective in the interview. Furthermore, their team provided recommendations for our wet lab team on the use of unique engineered strains such as Senecella to improve PHB production efficiency.

Takeaways

We learned that BluePHA’s bioplastic technology, which uses biomass or waste cooking oil to synthesize biodegradable PHA, provides an implementable pathway toward a circular economy. Furthermore, when conducting a life cycle analysis, we learned that upscaling a process is a complicated process with many real-world variables that could result in a wide range of values. Thus, an LCA has high utility in pinpointing optimizable steps of the process where values have a higher range. Beyond manufacturing, we noted that a successful LCA must take into account the impact of the gas and electricity used to transport and power the process, not just the carbon footprint of the manufacturing itself. Additionally, we learned that in industrial processes, yield is often low, and they shared the value of optimization techniques such as refining technologies and choosing the right feedstock to maximize yield to enhance profitability and productivity.

Implementation

We implemented the knowledge about metabolic engineering, life cycle analysis, and circular economies that we gathered during our correspondence with BluePHA in our dry lab and LCA. In conducting our LCA, we referenced BluePHAs papers as context, which concentrated on utilizing real-world production data to conduct a comprehensive analysis, and particularly learned from their example to emphasize the importance of factoring in downstream processes such as purification in calculating the environmental impact of our process. Following their example, we also pinpointed optimizable steps in our process such as transport and resource management. Furthermore, our takeaways about the importance of maximizing yield brought us to conduct a flux balance analysis to optimize our PHB synthesis. Using genes, enzymes, and reaction targets, we modified Pseudomonas Putida KT2440 to increase yields of NADPH and Acetyl-CoA, key metabolites in the pentose phosphate (PP) pathway.

Entrepreneurship

iGEM Entrepreneurship Summer School

Overview

Four of our team members attended the iGEM Startups Virtual Summer School, which featured speakers such as Johannes Sonnenschein, Head of Business Development at Insempra; previous Best Entrepreneurship prize winners like Santiago Plata Salazar and Arely Jiménez Hernández; and Florian Kroh, who led pitching workshops. The curriculum covered important components to consider when building a commercial venture from a scientific idea, including market validation, strategic feasibility, and effective communication.

Objectives

Our primary objective was to learn how to structure a compelling business proposal, as our team had limited prior experience in entrepreneurship and commercialization. We aimed to understand the essential components of a strong proposal and some strategies for communicating scientific ideas in a way that appeals to investors and partners. Furthermore, we sought to gain first-hand insights from previous competition winners and industry professionals, whose experiences guided how to refine how business models, anticipate potential market challenges, and position our project for long-term commercial success.

Takeaways

We learned that a successful venture is built on a clear mission addressing a defined problem with a specific solution, all supported by a unique value proposition. For example, we learned the necessity of conducting thorough research on our market, demonstrating feasibility with a cohesive manufacturing plan and exit strategy, and building credibility through thorough stakeholder and risk analysis. Also, we recognized the importance of aligning the project with Sustainable Development Goals (SDGs) and presenting all this information in a professional, clear, and structured manner.

Implementation

We used these teachings to structure our entire business plan and wiki pages. We formulated a precise Mission, Problem, and Solution statement and defined distinct Unique Value Propositions for our B2B and B2C customers. The framework guided our research, helping us identify what specific information we needed to find, such as market sizes and potential manufacturing partners. We outlined a manufacturing and exit strategy, and conducted a SWOT analysis to showcase strategic awareness, ensuring our project presentation is both professional and convincing. Moreover, we performed a Life Cycle Analysis (LCA) in order to quantify our project’s impact on the environment, allowing us to gain a more comprehensive understanding of our alignment with the SDGs.

GoodCycle Interview

Overview

We conducted an interview with 王佳韵 (Wang Chia Yun), the manager of the sustainability department at GoodCycle. GoodCycle is an environmental brand in China that strives to reduce plastic waste by collaborating with various organizations to collect plastic and then mechanically upcycle it into high-value consumer products like school uniforms and pencil cases.

Objectives

Our goal was to learn from an established industry player about the practical realities of operating a circular economy business. We chose GoodCycle because they have been working on reducing plastic waste in China since 2017, making them an experienced and reputable organization in the field of sustainable recycling. By consulting GoodCycle, we hoped to learn from their experiences about the process of degrading PET plastic and how they sustain large-scale plastic reduction initiatives. Understanding their production process of turning PET waste into consumer products can provide us with practical insights when we conduct our own project.

Takeaways

Through this interview, we gained a deep, practical understanding of the supply chain and the manufacturing operation of plastic upcycling. GoodCycle collected PET bottles from their collaborator, crushed them into bottle bricks, and sent them to a cleaning factory. The factory then removes the covers, sterilizes the bottles, shreds them into flakes, and melts the flakes, turning them into resin chips. Next, another factory in this chain makes polyester fibers from the molten plastic by forcing the plastic through a fine-hole spinner. Lastly, the cloth factory can produce uniforms out of the polyester fiber. Additionally, the interview provided valuable insights into how to effectively track and communicate environmental impact through metrics like CO₂ savings. They use the Greenhouse Gas Equivalencies Calculator invented by the United States Environmental Protection Agency to calculate how much carbon emissions they reduced.

Implementation

By designing and constructing novel biological parts, we aim to develop an innovative production that aligns with the principles of the circular economy, a concept that was emphasized by our interview with GoodCycle. Their mission of transforming PET into clothing for people in rural areas aligns with our goal to contribute to our society by reducing carbon emissions in the biomanufacturing industry. Therefore, their information on how PET is upcycled helped us to build the framework of our life cycle analysis. Specifically, understanding that the process requires separate facilities allowed us to define our LCA boundaries and identify the costs and energy consumption. This allowed us to quantitatively analyze whether our biological process offers a better sustainable alternative.

Education

Ben Kask Interview

Overview

We met with the instructional coach of our school, Ben Kask, to discuss how we can make the agar art curriculum handbook more effective. Ben Kask has years of teaching experience and provided practical advice on how to improve the usability and clarity of our handbook.

Objectives

Our objective was to revise our handbook while considering a teacher’s perspective, ensuring that the curriculum is easier for educators to implement effectively in a classroom setting. Since Ben Kask is the instructional coach in our school, it was relatively easy for us to gather valuable insights from him. His familiarity with our school’s teaching methods, students’ learning styles, and curriculum procedures in classes allowed us to receive targeted feedback that can be directly applied to our handbook. By consulting him, we hoped to make our handbook more practical and accessible to teachers and strengthen its alignment with real-world teaching environments, ensuring that the Agar Art workshop can be readily adopted by educators beyond our school.

Takeaways

We recognized that our strengths for this handbook are that the curriculum is engaging, creative, and informative. Meanwhile, experiments in synthetic biology are often lacking in the normal curriculum in international schools in China. Therefore, we recognized this project as a bridge in making synthetic biology more accessible to high school students, while also appealing to teachers such that they can implement this in their own curriculum. Nonetheless, our curriculum had certain aspects that could have been improved. For example, the connection between agar art and synthetic biology was not clearly explained, which could reduce the curriculum’s appeal. Besides, our arrangement of the pages was a little confusing to readers who got to our handbook for the first time. Therefore, we spent some time rearranging the pages.

Implementation

First, we emphasized our objectives and the relationship between agar art and synthetic biology, appealing to the value of this curriculum for teachers who might want to do a hands-on experience exploring the bacterial operons. Secondly, we made some edits to the font layout, bolding the content that can best summarize the main idea, making it more perceptible for users. We added captions below our animations to clarify the process. Moreover, we added bits of teaching techniques into the lesson plans and arranged the Appendix (lab instructions), connecting to the lesson plan, because the logic is more fluent when we combine them. To that end, to deepen our understanding of effective curriculum design and assessment, we reached out to Biobuilder, an organization that is more connected with synthetic biology.

Biobuilder Interview

Overview

We interviewed Chloe Franklin, the Education Program Coordinator of Biobuilder, who offered valuable suggestions to visualize the effectiveness of our program and create resources for potential users. Chloe Franklin is an expert in teaching and has extensive experience in aligning hands-on workshops with diverse educational standards.

Objectives

Our goal is to learn effective ways to structure our resources in order to maximize the teaching qualities and gather advice on improving workshop design, assessment tools, and teacher training. To achieve this, we connected with Biobuilder, a renowned organization dedicated to the field of synthetic biology. Since Chloe Franklin has extensive experience on the educational side of synthetic biology, we can learn about their teaching framework and thus guide us to refine our curriculum’s structure. Moreover, we aimed to explore a more effective evaluation method of the workshop, which can help us to revise the workshop to enhance its effectiveness in raising awareness about synthetic biology.

Takeaways

The key takeaway from this interview was learning how to align synthetic biology curricula with international standards, emphasizing the importance of clear learning objectives and providing ready-to-use teacher resources. We learned that workshops are more effective when guided by teachers with clear instructions, materials lists, and structured protocols; visuals should be paired with captions where necessary; and lab steps should be clearly numbered and spatially organized. Finally, Chloe Franklin gave us the standards for pre- and post-lab surveying, particularly emphasizing the utility of multiple-choice questions for the sake of quantitative analysis.f

Implementation

After the interview, we designed a pre- and post-survey for the workshop to measure how effectively the workshop enhanced students’ understanding of synthetic biology. We revised the objectives using measurable verbs and reformatted surveys into multiple-choice questions for easier scoring and analysis. Moreover, we developed PPTs for each of the modules included in the handbook, along with handouts and structured lab guides with clear visuals and captions.