▼ INTEGRATED HUMAN PRACTICES ▼

HUMAN PRACTICES

The St. Paul's College iGEM team is driven by a commitment to a responsible and meaningful rendition of the scientific method, along with an emphasis on consciousness of the social consequences of scientific projects . Our Integrated Human Practices ensures that our project is continuously shaped by expert insight and community engagement, moving beyond the lab to consider the broader implications of our work.

We proactively seek guidance through in-depth interviews with professors and researchers, grounding our technical approach in expert knowledge. To broaden our perspective, we have attended specialised workshops on bioethics and biosafety, ensuring that our work is compliant to the strictest standards.

Understanding that science thrives on collaboration, we have hosted exchange events to build a community of shared learning with fellow iGEMmers. Finally, we are passionate about inspiring the next generation by designing and leading educational workshops for younger students, demystifying synthetic biology and fostering a culture of inclusion within the scientific community.

This holistic approach is fundamental to our process. By listening, learning, and sharing, we ensure that not only is our project scientifically rigorous, but also ethically sound and socially responsible.

Immersive Experience

Our team believes in experiencing the problems we're trying to solve firsthand. This immersive approach helps us develop solutions that are grounded in reality and truly address the needs of the communities and environments we aim to serve.

A Deep Dive into Marine Plastic Pollution at SWIMS HKU

A core tenet of our iGEM journey is to ensure that our project is grounded in real-world problems. In the belief that we must step out of our lab so as to achieve a good understanding of the issues we are tackling, we led our team to the Institute of Marine Science (SWIMS) at the University of Hong Kong for an immersive workshop on Plastic Pollution and Marine Debris at the Cape d'Aguilar Marine Reserve.

Figure 1. A group photo at SWIMS, HKU.

Why This Workshop? The Integrated Human Practices Link

Our project is developing a biological solution to address plastic pollution. Before we can propose a solution, we must first deeply understand the problem not just on paper, but in reality. Attending this workshop was a fundamental part of our Integrated Human Practices, allowing us to learn directly from field experts and see the impact of microplastics on a local ecosystem firsthand. This knowledge is crucial for ensuring our project is relevant, effective, and responsible.

A Start with Theory and Wonder

The workshop began with an enlightening seminar from the SWIMS researchers. They detailed the devastating impact of marine debris and microplastics on local biodiversity, from the largest fish to the smallest zooplankton. This session moved the issue from an abstract concept in a textbook to an urgent, local crisis.

Following the talk, we were given a tour of the magnificent facility. We saw state-of-the-art lab appliances used to analyse marine samples and were fascinated by the biosamples of diverse marine organisms, each a piece of the complex puzzle that composes the Hong Kong marine ecosystem.

The highlight for many was the touch tank. Being able to gently handle sea urchins, corals, sea cucumbers, and crabs was a powerful reminder of what we are working to protect. It transformed these creatures from abstract "marine life" into tangible, fragile organisms whose survival is threatened by pollution.

Figure 2. Plastic sampling workshop at SWIMS, HKU.

A Process of Hands-On Discovery

The theoretical knowledge quickly turned into practical action. We moved to the beach outside the facility for a hands-on plastic sampling workshop. Armed with sieves and trays, we collected sand and sediment samples and learned the meticulous process of extracting microplastics.

Most importantly, we were taught the standardized four-category sorting system for plastic debris, a critical skill for accurate environmental monitoring:

Seeing the sheer volume and variety of plastic, especially the pervasive microplastics hidden in the sand, was a sobering and motivating experience.

The Final Stop: A Sobering Visit to Lap Sap Wan

The most inspiring part of our day was the final stop: a visit to the infamous Lap Sap Wan (rubbish bay in Cantonese). We had prepared ourselves for a sobering sight of a shoreline choked with plastic. However, we were met with a powerful and hopeful revelation: the condition at Lap Sap Wan has improved dramatically.

Our guides at SWIMS explained that this visible improvement is the direct result of sustained conservation efforts, including organised clean-up drives and improved waste management policies. While our sieving activity revealed that microplastics remain a persistent and hidden challenge, the dramatic reduction of large-scale debris is undeniable proof that human action can and has made a tangible difference.

Bringing It All Back to the Lab

This workshop was more than just a field trip; it was a pivotal moment for our team. It:

• Provided Critical Context: We now have a firsthand understanding of the scale and complexity of plastic pollution.
• Informed Our Design: Seeing the different categories of plastic waste helps us realistically scope our project's capabilities and target the most impactful areas.
• Fueled Our Motivation: Connecting with the marine life at SWIMS and witnessing the pollution at Lap Sap Wan renewed our determination to develop a meaningful solution.

We are immensely grateful to the dedicated team at SWIMS HKU for their time and expertise. The insights gained from this day are directly integrated into our project design, ensuring our work in the lab is always connected to the real world it aims to improve.

Academic Outreach

Refining Our Project with Prof. Nathan Crook from North Carolina State University

A pivotal part of our iGEM journey involves seeking expert advice to strengthen the scientific rigour and practical feasibility of our project. We are therefore incredibly fortunate to secure a virtual meeting with Professor Nathan Crook, a leading expert in metabolic engineering and synthetic biology from North Carolina State University. The discussion provided invaluable, actionable insights that are directly shaping the next phase of our wet-lab work.

The Core Philosophy: Understanding Microplastic Breakdown

Professor Crook began by reframing our objective. He emphasised that the breakdown of microplastics is not simply about making them "disappear." Instead, it is a process of transformation: converting large polymers into smaller molecules, primarily through physical, chemical, and biological means. This crucial perspective ensures that we are focused on tracking the complete journey of plastic degradation, not just its initial fragmentation.

Figure 3. An online zoom consultation with Professor Nathan Crook from the department of Chemical and Biomolecular Engineering at NCSU.

Actionable Lab Strategies and Experimental Design

The meeting was immensely practical, yielding specific protocols and ideas for our laboratory investigations:

1. A Phased Experimental Approach: Prof. Crook advised a clear, step-by-step validation process. Before testing our engineered enzymes on plastic beads, we should first prove our experimental setup works using normal E. coli and a standard assay. This establishes a baseline and confirms our methods are sound before introducing more complex variables.
2. Innovative Bioreactor Design: We discussed a key challenge: how to contain our engineered cells while allowing them to act on plastic waste. He suggested a sophisticated bioreactor design featuring a physical filter to retain our bacterial cells. A medium containing PET microplastics could then be circulated over this immobilised cell bed, allowing for continuous, contained, and efficient partial depolymerisation of PET.
3. Novel Detection Methods: A significant breakthrough from the discussion was on how to detect successful degradation. Prof. Crook introduced advanced ideas beyond standard assays:
   • A Forensic Analog: Using a His-tagged version of our PETase enzyme to visually track its binding and activity on plastic surfaces under microscopy.
   • Colorimetric Biosensors: Exploring the use of biosensors specifically designed to detect Terephthalic Acid (TPA), the primary breakdown product of PET. Alternatively, using a compound like p-Nitrophenyl butyrate (pNPB) which, when cleaved by esterases like PETase, changes color to yellow, providing a clear and quantifiable visual signal of enzyme activity.

Inspiring the Next Generation

Beyond the technical details, we also discussed the importance of nurturing future scientists. Prof. Crook shared his thoughts on how to encourage wet-lab participation among new junior members. His key advice was to design onboarding experiments that are visually engaging (like color-changing assays) and have a high chance of success, providing immediate reward and fostering a passion for hands-on science, a principle we are actively integrating into our team's mentorship structure.

Integrating Expert Advice into Our iGEM Journey

This meeting was a perfect example of Integrated Human Practices in action. The insights from Prof. Crook are not just notes in a lab book; they are being actively translated into our experimental plans:

• Revising our validation pipeline to include control experiments with non-engineered E. coli.
• Drafting designs for a more effective bioreactor prototype.
• Investigating the implementation of His-tagging and colorimetric assays (pNPB) for more robust detection of our enzyme's activity.

We are deeply grateful to Professor Nathan Crook for his generosity, time, and exceptional guidance. His expertise has provided our project with a new level of depth and direction, exemplifying the collaborative spirit that drives scientific discovery forward.

Professor Michael Chan - Materials Support and Experimental Guidance

Materials Provision:

Professor Chan has generously provided our team with essential chemical reagents to advance our experimental work. These include:

• PBS (Phosphate Buffered Saline) solutions for maintaining physiological pH conditions in our enzymatic assays
• DMSO (Dimethyl Sulfoxide) solvent for dissolving and handling organic compounds
• pNPB (p-Nitrophenyl Butyrate) substrate for conducting colorimetric assays to measure esterase activity

Advanced Imaging Opportunity:

We have been granted access to scanning electron microscopy facilities scheduled for September 19, 2025. This will enable us to perform high-resolution visualisation of PET surface degradation and directly observe the morphological changes induced by our engineered enzymes.

Figure 4. SEM imaging at Queen Mary Hospital with Mr. Mark Szeto

Methodological Guidance:

Professor Chan provided crucial statistical recommendations to strengthen our experimental approach. He emphasised the importance of implementing rigorous statistical tests to determine the significance of performance improvements in our modified enzymes. Specifically, we will now focus on quantifying:

• Statistical significance in PET degradation rates between wild-type and engineered enzymes
• Quantitative improvements in substrate binding affinity through appropriate statistical validation

This mentorship has significantly enhanced both our technical capabilities and experimental design, providing us with essential tools and methodologies to rigorously validate our enzyme engineering progress.