Overview

Throughout this year, we learned a lot from the work of previous iGEM teams and scientific research. Their experiences helped us think beyond just building an enzyme. Step by step, we connected different aspects of our project, from experiments and data measurement to education and economic analysis, into one complete system.

Our goal was simple: to make something useful, understandable, and lasting. We designed biological parts that can be reused, built a CO₂-based measurement method that other teams can easily follow, created education programs for all ages, and developed an economic plan.

By sharing these contributions, we hope future teams can use what we built, improve it, and continue exploring new ways to connect synthetic biology with people.

Parts

BBa_25CCXO78 (Basic part, coding)

endo-β-1,4-glucanase gene from Bacillus subtilis JA18.

This basic coding part expresses the endo-β-1,4-glucanase gene found in Bacillus subtilis JA18. This enzyme specializes in breaking the internal β-1,4 glycosidic bonds in cellulose, cutting long chains of polysaccharides into shorter pieces and eventually turning them into fermentable sugars. We cloned this gene from the JA18 strain and studied its chemical properties, including the best pH and temperature for its activity, through various literature reviews. The enzyme works really well under moderate temperature and pH conditions. We used this part to express endo-β-1,4-glucanase for food waste pre-treatment. However, this construct could also be applied by other iGEM teams to produce endo-β-1,4-glucanase for various purposes, such as agricultural, environmental, or research purposes.

BBa_25P3Y9PI (Composite part, coding)

YebF-endo-β-1,4-glucanase

This composite part encodes a fusion construct combining the Bacillus subtilis JA18 endo-β-1,4-glucanase with E. coli K12 secretion carrier protein YebF. In this composite design, YebF directs export of the fusion to the periplasm and culture supernatant, where the enzyme endohydrolyzes internal β-1,4-D-glycosidic bonds in cellulose. Our team utilizes this part to pretreat food waste, enhancing the black soldier fly larvae's degrading efficiency. However, this part holds potential beyond our project and can also be used by other iGEM teams. This part could also be utilized for agricultural and environmental purposes. By demonstrating how YebF can effectively transport proteins to the extracellular space, other teams can use a similar construct to secrete their gene of interest into the periplasm into the extracellular space of E. coli, serving as a platform for future teams who seek to express enzymes in large quantities, reducing the costs of cell lysis and downstream purification.

Measurement

This experiment is a reproducible CO₂-based attempt for evaluating anaerobic organic waste degradation under controlled conditions. By applying the Gompertz function uniformly across scenarios, we were able to estimate comparable kinetic parameters and identify the strengths and limitations of using standard growth models for various substrates. Our results provided numerical baselines such as the asymptote, lag time, and growth rate that can serve as benchmarks for assessing future studies involving improvements using enzymes such as endo-β-1,4-glucanase activity.

In addition, the strong fit observed in control samples compared to the weaker fits for fruit substrates has provided insights into how substrate complexity and fiber content affect model performance. This reveals the limits of classical kinetics for heterogeneous materials. With the steep decline accounted for around 150,000 seconds, the study also showed a good example of validation of data quality in a small-scale experiment.

In conclusion, this experiment contributes new modeling insights, a reproducible CO₂-based protocol without the use of advanced instruments, quantitative benchmarks for enzymatic improvement, and a transparent approach to data validation. Overall, this work demonstrated that a familiar biological observation has been transformed into a quantitative, reproducible, and open-source measurement framework that future teams can use to evaluate and improve biodegradation efficiency.

Education

Our project's education plan provides a practical and reproducible framework for future iGEM teams to design and implement outreach and educational initiatives. Our programs cover a wide range of age groups from kindergarteners, elementary students, junior and senior high school students, to elders. On top of this, we have considered international communities in Sri Lanka and Vietnam.

Our tools are accessible such as the eight-panel comic with coloring activities, the board game, Scratch animations, teaching card decks, problem-solving games and large-font visuals with oral storytelling. We developed a framework accessible to most age groups and English Language Learner (ELL) friendly. This is because we want the future iGEM groups to continue to consider and advocate for accessible learning resources.

In addition, we incorporated team-based and interactive visuals to make biology accessible with limited resources. These educational activities introduced the public to synthetic biology, enzyme engineering, black soldier fly larvae applications, and food waste solutions, fostering curiosity, critical thinking, and intergenerational engagement.