Contribution

Parts

In our biobricks, we contributed 2 enzymes PKS and AGS2, 3 different anchor efficiency determination plasmids, and 3 improved proteins CWP-VHb, Sed-VHb, and Tip-VHb.
PKS is a gene related to melanin synthesis, and AGS2 is a gene related to pullulan synthesis. We successfully obtained and knocked out these genes, significantly increasing the yield of β-glucan in Aureobasidium melanogenum BZ-11. Additionally, we selected 3 cell wall anchor proteins as targets, each connected to GFP (green fluorescent protein), identified the most effective anchor protein, and further fused it with Vitreoscilla hemoglobin (VHb) to enhance the oxygen uptake capacity of the chassis organism, thereby improving production efficiency.

BBa_K5314001: PKS is an enzyme with multiple biosynthetic activities, including the melanin synthesis that we are interested in. In Aureobasidium melanogenum BZ-11, it regulates the first step of melanin synthesis, which is the formation of 1,3,6,8-THN from acetyl-CoA and malonyl-CoA. Therefore, PKS is a key enzyme in melanin synthesis. After we knocked out this gene, we downregulated the biosynthesis of melanin.

BBa_K5314002: AGS2 is an alpha-glucan synthase, primarily involved in the biosynthesis of pullulan, playing a role in catalyzing carbon chain elongation and transmembrane transport; it is a key enzyme in pullulan synthesis. After we knocked out its gene, we downregulated the biosynthesis of pullulan.

BBa_K5314006, BBa_K5314007, BBa_K5314008: We connected EGFP to the cell wall anchor proteins we identified and expressed EGFP fluorescent protein in Aureobasidium melanogenum BZ-11 to display the anchoring efficiency through EGFP. Eventually, we found the most effective set connected to the target protein VHb, aiming to enhance the oxygen uptake capacity of the chassis organism.

BBa_K5314003 BBa_K5314004 BBa_K5314005: We designed and predicted the outcomes of 3 different types of VHb fusion cell wall anchor proteins, aiming to explore the effect and anchoring efficiency of connecting different types of cell wall anchor sequences to VHb protein. By predicting the structures, we could evaluate the effectiveness of different anchor proteins.

Table 1: New Parts Table

Hardware

For society, our precision feeding hardware is specifically designed to address key issues in aquaculture, directly contributing to the entire aquaculture industry. This hardware significantly enhances the economic efficiency of current aquaculture practices while promoting environmental sustainability. By detecting and analyzing feed sedimentation, we designed and manufactured a device that reduces feed waste and water pollution. This innovation not only significantly improves feed utilization but also protects the ecological environment by reducing water pollution from continuous feeding, ensuring water health, which has significant economic value and environmental protection significance for aquaculture farmers.

Globally, aquaculture is a crucial part of the entire food supply chain. With the growing global population and increasing demand for aquatic products, our precision feeding device has broad application prospects and can be promoted worldwide, helping more aquaculture owners achieve efficient and sustainable production methods. By reducing feed waste and improving water quality, our device contributes to the sustainable development of the aquaculture industry. This also aligns with the United Nations Sustainable Development Goals, particularly Goal 2 (Zero Hunger), Goal 6 (Clean Water and Sanitation), Goal 12 (Responsible Consumption and Production), and Goal 14 (Life Below Water).

For the iGEM community and the scientific community, as an innovative part of the iGEM project, our hardware design demonstrates how engineering technology can solve real biological engineering problems. Our work involves the entire process of hardware design, 3D modeling, experimental verification, and improvement, showcasing the complete engineering process from problem identification to solution. We provide detailed design ideas, manufacturing blueprints, and improvement suggestions, which will be of great reference value to other teams in the iGEM community and the entire scientific community, inspiring more teams to innovate and practice in the field of aquaculture.

We firmly believe that through our relentless efforts in hardware, we can help farmers improve feeding efficiency and environmental quality, and inspire more iGEM teams and researchers to jointly promote the development of biotechnology and environmental science. Our precision feeding device demonstrates how innovative thinking and advanced technology can solve real biological and environmental problems, which is a positive embodiment of the iGEM spirit.

Figure 1: Hardware

Model

Using the Carveme automated reconstruction tool based on Linux, we constructed the whole genome network of Aureobasidium melanogenum P16 and manually corrected parts of the network, greatly aiding future teams that use Aureobasidium melanogenum as a chassis organism and need its whole genome network. Through this work, we not only provided innovative tools and methods to solve practical laboratory problems but also laid important foundational data for the scientific community to understand and apply this microorganism.

At the societal level, our metabolic model significantly accelerated the research and production optimization of β-glucan. By identifying key gene knockout targets, we improved experimental efficiency and result reliability, potentially promoting the widespread application of these high-value compounds in medicine, food, and materials, achieving greener and more efficient production models, benefiting society.

Globally, our research provides new ideas and tools for the synthetic biology community, promoting the development of basic science and offering potential solutions to global challenges such as resource shortages and environmental pollution. By optimizing gene networks and protein structures, our work lays a solid foundation for the development of microbial synthetic biology, potentially driving the realization of more innovative applications.

For the iGEM community, our contributions are particularly significant. From whole genome metabolic networks to local metabolic networks, we ultimately identified target genes to be knocked out, further narrowing the experimental target range and improving experimental efficiency. This model not only optimizes β-glucan production but also provides valuable data support for future research. Our metabolic model provides validation and reference for other iGEM teams studying the synthetic pathways of β-glucan, melanin, pullulan, etc., greatly reducing trial and error costs and improving experimental efficiency. Additionally, the molecular model we constructed demonstrates innovative pathways for combining cell wall anchoring sequences with Vitreoscilla hemoglobin, enabling VHb secretion to the extracellular environment, improving oxygen transport efficiency, and revealing the impact of C-terminal cell wall anchoring sequences on protein activity. This discovery not only provided direction for our team's experimental optimization but also offered valuable experience and inspiration for other iGEM teams in optimizing protein structure and performance, such as mutating amino acids or adding linkers to optimize protein conformation.

Overall, our Model work showcases the importance of interdisciplinary integration and innovative thinking in synthetic biology, contributing significantly to the iGEM community's knowledge, data, and tool repository. We hope to share our findings and experiences with community members, jointly promoting the frontier exploration and practical applications of synthetic biology. Through such knowledge sharing and technological innovation, we expect every team to benefit, pushing global synthetic biology development to new heights.

Figure 2: Model

integrated HP

We conducted extensive integrated HP with various societal sectors, including β-glucan producers, feed manufacturers and buyers, beauty and cosmetics institutions and customers, pet hospitals, aquariums and zoos, farms, medical personnel and patients, university professors, government officials, students from various fields, human food suppliers, and more.

For society, we provided pharmaceutical manufacturers with new drugs to produce and sell, helping fill their productivity gap after the antibiotic ban and providing more jobs for society. In our communications with feed manufacturers and farms, as well as aquariums and zoos, we promoted solutions to enhance animal immunity, improving feed quality in the livestock industry, and introducing new antibiotic substitutes to farms, aquariums, and zoos, reducing reliance on antibiotics. We provided new products with unique effects to beauty and cosmetics institutions, customers, and pet hospitals. Through interactions with medical personnel, patients, and the public, we raised awareness of antibiotic misuse issues and alternative solutions, helping reduce societal demand for antibiotics, thereby protecting human and animal health. By organizing public dissemination activities such as online podcasts, public lectures, and debates, we educated students and the public about β-glucan and ethics, increasing global understanding of our project and promoting ethical reflection.

Globally, our project significantly advanced the achievement of the United Nations Sustainable Development Goals (SDGs). Specifically, we supported Goal 3 (Good Health and Well-being), Goal 8 (Decent Work and Economic Growth), Goal 9 (Industry, Innovation, and Infrastructure), Goal 10 (Reduced Inequalities), Goal 11 (Sustainable Cities and Communities), and Goal 14 (Life Below Water). By collaborating with various global stakeholders, we reduced the generation of antibiotic-resistant bacteria, promoting global public health. We also proposed The TPNR Comprehensive Synthetic Biology Ethics Framework, providing a systematic ethical guidance framework for global synthetic biology research, ensuring that project development considers technological advancement and social, environmental, and ethical impacts. This framework promotes the coordinated progress of global science and ethics.

For the iGEM community, we developed and applied the TPNR (Theory-Practice-New Theory-Re-practice) framework, offering other iGEM teams an integrated model of theory and practice, continuously optimizing and innovating human practice. Through close collaboration with multiple universities and iGEM teams, we engaged in in-depth exchanges in experimental techniques, model construction, project management, and ethical discussions, enhancing overall research levels and providing valuable data and knowledge to the community.

We firmly believe that through these iHP efforts, we not only advanced our project but also made significant contributions to society, the world, and the iGEM community, continually moving forward with the original intention of making the world a better place through synthetic biology.

Figure 3: integrated HP

Education

We've undertaken voluminous, all-encompassing, and profound data analysis in the realm of education, dedicating ourselves to a detailed exploration of participation and educational scenarios since the dawn of iGEM. Our analysis unveiled imbalances in engagement and thus reflected iGEM educational inequalities across various facets and echelons, shattering the misconception that educational standards are in lockstep with economic prosperity, thus breaking free from the deadlock of misinformed, haphazard educational initiatives.In response to this, we innovatively introduced the GENE Cycles, a methodology that places great emphasis on the formulation and investigation of specific issues. This significantly enhanced the pertinence and effectiveness of our educational outreach. Following exhaustive preliminary research, our diverse, engaging, and regionally tailored educational activities, designed to address the unique needs of different age groups and issues, garnered commendable feedback and yielded remarkable results. Not onlt did We successfully nurture the nascent forms of new iGEM groups in kinds of schools that had previously shown low participation rates, sharpening the focus of our educational activities but also did we extend our reach, initiating outreach in Oceania, a region where only two teams existed. Furthermore, the constructive feedback we received was of immense value. By integrating the GENE Cycle, we continuously refined our approach, building on past experiences.Last but not least, We expanded the scope of our educational efforts, initiating a dialogue with downstream sales producers, users, and regulators. This not only solidified the tangible impact of our product contributions to the world but also lowered barriers to the future promotion of synthetic biology products, endeavoring to dispel the public's misconceptions regarding their prohibitive costs and complex application.

Figure 4: Education