In our project, Integrated Human Practices serve as a critical bridge between laboratory innovation and real-world needs, guiding us to deeply explore the challenges of chitin resource utilization and refine our solutions to achieve both ecological and economic value.

Figure 1 Integrated Human Practices Mind Map
Chitin, the second most abundant biopolymer on Earth, is widely distributed in shrimp shells, crab shells, fungal cell walls, and insect exoskeletons. However, its rigid structure makes it difficult to degrade naturally, leading to massive waste accumulation—especially in aquaculture and seafood processing industries—while its high-value derivatives (e.g., chitooligosaccharides, N-acetylglucosamine) remain underutilized. Meanwhile, natural chitinase production is low, limiting large-scale industrial application. To address this, we aim to construct a high-efficiency chitinase expression system using synthetic biology, enabling efficient degradation of chitin waste and resource recycling.
Through human practices, we seek to answer three core questions:
1.What are the actual needs of stakeholders in different areas for chitinase technology?
2.How can we optimize our genetic engineering system to align with industrial application standards (e.g., enzyme activity, stability, cost)?
3.How to raise public awareness of chitin resource recycling and promote the adoption of this green biotechnology?
By integrating insights from questionnaires, expert interviews and visits, we continuously adjust our experimental design—from screening chitinase genes to optimizing expression conditions—and shape our product positioning, ensuring our innovation not only advances science but also solves practical societal problems.

We have designed two questionnaires for our project. One questionnaire focused on investigating the public's awareness of chitinase and the protection of marine pollution, while the other focused on the market application and promotion of chitin and chitinase. The following is our analysis of the results of these two questionnaires.
From these two questionnaires, we have also obtained some ideas and suggestions regarding subsequent education activities.

2.1 Questionnaire 1: Public's awareness of Chitinase and the protection of marine pollution
2.1.1 Analysis of the educational level of respondents
Of the 468 respondents who participated in the questionnaire, bachelor's degree was the highest (44.66%), followed by secondary school and below (22.44%), college (21.37%), and master's degree and above was the lowest (11.54%).

Figure 2 Respondent's Education Level
This indicates that the respondents of this questionnaire are mainly concentrated in the population with undergraduate education and below, and the results of the study may reflect the cognitive level of the general public more than the views of experts in high-end professional fields. This also suggests that this questionnaire has strong generalizability and can reflect the preliminary knowledge of people with different educational backgrounds about chitinase in a more comprehensive way.
Based on the age distribution of the respondents as shown in the questionnaire, we propose some possible strategies for popularizing the knowledge. The low-education group has less knowledge about chitinase, and needs to adopt more understandable ways of popularization, such as graphic explanations, vivid cases, and popular science videos. Since the proportion of people with bachelor's degree is the highest, this group can be targeted to provide a slightly more in-depth introduction of the technology and application level, such as the actual application cases of chitinase in biomass degradation, agriculture, medicine and other fields.
2.1.2 Age analysis of respondents
a.Percentage of each age group

Figure 3 Respondent's Age Distribution
According to Figure 2 above, the highest percentage (46.58%) was found in the 26-45 age group, followed by 46 years old and above (32.05%), 13.89% was found in the under 18 age group, and the lowest percentage (7.48%) was found in the 18-25 age group. This result shows that the main audience of the survey is the young and middle-aged group (26-45 years old), which may be involved in biotechnology, agriculture, environmental protection, and other industries, and will be involved in the related knowledge and applications. In general, the age distribution is more in line with the age distribution in the current society, which also shows that this questionnaire is universal and representative, and more valuable for reference.
b.Relationship between age and their awareness level
People aged 26-45 years old are likely engaged in industries related to agriculture, biotechnology, etc. Therefore, they may have a better understanding of chitinase or pay more attention to its application in resource degradation and transformation.
People aged 46 years old or above may be engaged in management, decision-making and other positions, and concerned about the application of technology in industry, so they may be more concerned about the market prospect and commercialization value of chitinase.
Young people under 25 years old are mainly students. They are less involved in practical applications, but may have been exposed to the concept of chitinase in biology or environmental science courses.
We have collected some ideas for popularizing the concept in different age groups. For people over 25 years old, we can introduce the application of chitinase in bio-degradation, bio-energy, agriculture and food processing, and emphasize its economic value. For students aged 18-25 years old, we can provide academic lectures, such as the mechanism of action of chitinase, the construction of genetic engineering expression system, etc., to enhance their professional interest. For teenagers under 18 years old, we can use fun and educational methods, such as interesting animations and experimental demonstrations, to enhance their interest in biotechnology.
2.1.3 Basic understanding of chitinase

Figure 4 Public Basic Understanding of Chitinase
According to Figure 3, among the 468 respondents, 30.56% understood the specific role of chitinase, accounting for only a small proportion. At the same time, most of the remaining people cannot make accurate judgments about the specific efficacy of chitin, indicating that chitinase is relatively unknown in the public. This shows that our publicity about chitinase has a lot of space. By publicizing the use and advantages of chitinase in various aspects to consumers, the market of chitinase products can be further expanded.
2.1.4 Access to chitinase

Figure 5 Public Access to Chitinase
According to Figure 4, among 179 people who know the function of chitinase, about half of them know the information about chitinase through the Internet or social media, which is very prominent. In our era of rapid information development, social media is the main channel for information dissemination. In addition, about 35% of the respondents obtain relevant information through friends and family or books and magazines, highlighting the importance of personal social networks and traditional media in information dissemination. Nearly 20% of people learned about chitinase through television or school, indicating the diversity of information sources. Given these data, we can prioritize spreading them through social media.
Given these analyses, we can prioritize promoting our products through social media. By combining social media with traditional media (television, books) and interpersonal communication, we can increase public understanding and acceptance of chitinase.
2.1.5 Public awareness of marine pollution

Figure 6 Public Awareness of Marine Pollution
Figure 5 illustrates public awareness regarding the severity of ocean pollution. Unsurprisingly, over half of the respondents (65.81%) recognize that ocean pollution is becoming increasingly severe, indicating a favorable market environment for our product. Given that the threat to our daily lives is widely acknowledged, our product is expected to gain broad support. Additionally, while 25.85% of respondents have only partial knowledge about ocean pollution, they still represent potential advocates for chitinase-based solutions. Furthermore, although 8.34% of respondents have limited understanding of the issue, our team believes that the majority of them would be willing to join the community and contribute after targeted advertising and promotional efforts, which aligns with our goals and aspirations.
2.1.6 Public awareness of marine pollution

Figure 7 Distribution of Publicly Known Biomass Resources
Figure 6 reflects the public's familiarity with various types of biomass resources. Surprisingly, most respondents have some knowledge of relevant resources, whether in the categories of plant waste, animal waste, or marine resources. Plant waste is the most widely recognized, with 73.72% of respondents familiar with it. Only 14.96% of respondents are unaware of what biomass resources are. Given this context, we are confident that utilizing chitinase to extract biomass resources will be a well-received approach, as these resources are already familiar to the public.
2.1.7 Public attitudes towards the application of biomass resources to livelihoods

Figure 8 Public Attitudes Towards the Application of Biomass Resources to Livelihoods
According Figure 7, 95.51% of people believe that biomass resources are important in their daily lives. On the contrary, only 4.49% of people do not believe that biomass resources are important in daily life. From this, we can understand the public affirmation of biomass resources, and thus understand the importance of converting biomass resources for daily life.
2.1.8 Public perception of chitin applications

Figure 9 Public Perception of Chitin Applications
The survey shows that 65.38% of people believe that chitinase is of great help in alleviating marine pollution, while only 4.27% of people do not believe that chitinase is of great help in alleviating marine pollution. Another 30.34% of people are not very familiar with the specific role of chitinase. From this, we can see that most people have affirmed the effectiveness of chitinase in alleviating marine pollution, and also affirmed the significance of our project.
2.1.9 Public attitudes toward chitinase applications

Figure 10 Public Attitudes toward Chitinase Applications
Research shows that 88.03% of people maintain a neutral or neutral attitude towards products processed with chitinase, while only 11.97% of people are not very accepting of products processed with chitinase. By understanding the public's acceptance of products processed with chitinase, we believe that products processed with chitinase will be widely recognized by most people and gain a broad market.
2.1.10 Conclusion
This questionnaire indicates that the respondents are primarily young and middle-aged adults with bachelor's degrees or lower, reflecting that the results are more representative of the general public than of specialized experts. At present, public understanding of the specific functions of chitinase remains relatively limited; however, there is widespread recognition and concern regarding the importance of marine pollution and biomass resources. A majority of respondents acknowledge the potential application of chitinase in mitigating marine pollution and biomass conversion, and express acceptance or openness toward related products.
In terms of information dissemination, the internet and social media are the main channels through which the public accesses relevant knowledge. To further improve awareness and acceptance, tailored communication strategies—such as visual popular science for the general public and in-depth case studies for specialized audiences—are recommended. Overall, chitinase technology shows a solid foundation of public recognition and promising market prospects in the fields of environmental protection and biomass utilization.
2.2 Questionnaire 2: The Market Application and Promotion of Chitin and Chitinase
To understand public perceptions, market demands, and potential barriers to chitinase technology, we designed an anonymous online questionnaire distributed via online platforms (e.g., WeChat) and offline channels (e.g., schools, communities).
The ques aimed to answer key questions for our project:
1.What is the public's current awareness of chitin, chitinase, and chitin waste recycling?
2.What are the expectations for chitinase-derived products in different application scenarios (e.g., agriculture, food processing, environmental management)?
3.What obstacles might hinder the promotion of chitinase technology, and how can we address them through education or optimization?
2.2.1 Respondent demographics

Figure 11 Responses Overview

Figure 12 Age Distribution

Figure 13 Occupation Distribution
We collected 212 valid responses, covering diverse age groups and occupations:
The age distribution of the respondents is predominantly composed of individuals aged 1–17 years, accounting for 53.3% of the total, followed by those aged 46 years and above at 17.9%. The 18–25 age group represents 12.7%, while those aged 36–45 and 26–35 constitute 8.9% and 7.1%, respectively. In terms of occupation, students form the largest group, making up 63.2% of the respondents, followed by enterprise employees at 20.7%. Civil servants account for 5.2%, scientific and educational personnel represent 4.7%, and other occupations comprise 6.1% of the sample.
This diversity allowed us to analyze perspectives across different stakeholder groups, from potential end-users (e.g., students, families) to industry-related professionals (e.g., enterprise employees, civil servants).
2.2.2 Low public awareness of chitin and chitinase—a call for targeted education
The survey reveals a generally low level of public awareness regarding chitin and chitinase. Overall, 58.0% of respondents reported that they had never heard of chitinase, with significant disparities across age groups: while 76.3% of people aged 46 and above were unfamiliar with it, the proportion dropped to 51.3% among respondents aged 1–17. In terms of chitin awareness, only 53.1% of adolescents and 25.0% of enterprise employees knew that chitin is present in seafood waste such as shrimp and crab shells.

Figure 14 Heard of Chitinase

Figure 15 Environmental Concern

Figure 16 Chitin in Seafood

Figure 17 Channels to Learn Chitinase
These findings highlight an urgent need for targeted science communication. Given the strong preference for short-form videos as a medium for science outreach—expressed by 86.3% of all respondents and 88.8% of students—we plan to produce engaging video content tailored to different age groups. Animated explanations aimed at teenagers will illustrate the role of chitinase in waste recycling, while case-based narratives will be designed for adult audiences to enhance public understanding and promote broader dissemination of related scientific knowledge.
2.2.3 Strong support for enzymatic waste treatment—alignment with environmental goals

Figure 18 Key Functions

Figure 19 Related Fields
The results indicate a strong positive reception toward the chitinase-based method for seafood waste treatment, which received the highest approval rating among the options presented. Specifically, 53.3% of respondents awarded it a top score of 5/5, significantly outperforming traditional landfilling, which received the highest rating from only 29.7% of participants. This high level of acceptance is closely associated with its perceived advantages: 76.9% of respondents identified "environmental friendliness" as the most prominent benefit, followed by "resource recycling", which was recognized by 62.9% of those surveyed. In conclusion, these findings demonstrate strong public support for enzymatic waste treatment—a approach well-aligned with growing environmental goals.
Public recognition of environmental benefits reinforces our focus on improving the efficiency of chitinase in degrading shrimp/crab shells. We will prioritize optimizing enzyme activity under real-world conditions (e.g., varying pH, temperature) to align with industrial waste treatment needs, as supported by stakeholder feedback from environmental companies.
2.2.4 Clear Market Demand for Chitinase-Derived Products—Guiding Product Positioning

Figure 20 Acceptance of Waste Tech

Figure 21 Enzymatic Advantages
Survey results indicate a clear preference among respondents regarding application scenarios for chitinase-based products, with 81.1% prioritizing "degradable packaging" and 63.7% showing interest in "household cleaning" applications. Notably, student and enterprise employee groups demonstrated strong engagement in food-related uses such as food packaging, supported by 58.9% of students. When it comes to price sensitivity, 52.4% of participants would only consider purchasing chitinase-derived products if they were priced equally to conventional alternatives, while 20.2%—mainly students—would require at least a 10% price reduction to motivate purchase.
To meet market expectations, we will collaborate with food processing enterprises to develop cost-effective chitin-based packaging materials. Additionally, we will explore low-cost production strategies (e.g., optimizing fermentation conditions) to reduce prices, addressing the price sensitivity highlighted by the survey.
2.2.5 Barriers to Promotion—A Roadmap for Stakeholder Engagement
Implications for Stakeholder Collaboration:
With scientific/educational personnel: Co-develop educational materials (e.g., experiment kits for schools) to explain chitinase's mechanism.
With enterprises: Partner to showcase pilot projects (e.g., seafood waste recycling lines) as "application cases" for public outreach.
With civil servants: Advocate for policy support (e.g., subsidies for green technology promotion) to address cost barriers.

Figure 22 Enzymatic Advantages

Figure 23 Waste Utilization

Figure 24 Acceptable Recycled Products

Figure 25 Willingness to Purchase Marine Eco-Products

Figure 26 Expected Application

Figure 27 Willingness to Pay
According to the survey results, the key barriers to the adoption of chitinase technology were clearly identified: 83.0% of respondents considered "insufficient public awareness" to be the major obstacle, followed by "immature technology" (52.8%) and "high cost" (45.3%). In line with these challenges, significant educational needs were also expressed—65.1% of participants indicated a desire to learn about the basic principles of chitinase, while 62.7% showed strong interest in real-world application cases, such as how chitinase can convert shrimp shells into fertilizers.
With scientific/educational personnel: Co-develop educational materials (e.g., experiment kits for schools) to explain chitinase's mechanism.
With enterprises: Partner to showcase pilot projects (e.g., seafood waste recycling lines) as "application cases" for public outreach.
With civil servants: Advocate for policy support (e.g., subsidies for green technology promotion) to address cost barriers.
2.2.6 Conclusion
This survey provided critical insights for our project's iterative development:
1.Education: Use short videos and school collaborations to improve public awareness, focusing on young people and industry professionals.
2.Experiment Optimization: Prioritize enzyme efficiency in waste degradation and cost reduction to meet market price expectations.
3.Stakeholder Engagement: Address barriers through targeted collaborations with enterprises, educators, and policymakers.
By integrating these findings, we aim to ensure our chitinase technology is not only scientifically robust but also socially accepted and commercially viable.
The questionnaire analysis, as the core outcome of the project's research phase, has clearly outlined the practical pain points in chitin resource utilization, the current state of public awareness, and potential market demands, laying a data foundation for subsequent work. Building on this, we entered the stage of designing solutions—through in-depth interviews and on-site visits with academic experts, industry practitioners, and investment professionals—to transform abstract research data into specific technical paths and implementation strategies. The Integrated Human Practices at this stage serve as a key link connecting problem identification and solution implementation, driving us from "identifying problems" to achieving substantive breakthroughs in solving problems. The following will elaborate on how these stakeholder interactions have gradually shaped our technical optimization direction, application scenario selection, and business logic construction, forming an interlocking chain of solution design.

3.1 Interview with Processor Wang Shoubing
3.1.1 Expert Background
Professor Wang Shoubing from Fudan University. He serves as the director of the Environmental Ecology and Nature Conservation Professional Committee of the Shanghai Society of Environmental Sciences and other academic part-time positions. He actively participates in social services and academic exchanges, playing an important role in promoting the academic development and practical application of environmental science.

Figure 28 Interview with Professor Wang
3.1.2 Key Points
The purpose of this interview is to clarify the significance of our project and its contribution to society. Before the interview, our team prepared a discussion outline based on the role of chitinase in environmental pollution prevention and biomass resource conversion. We hope to receive valuable opinions and guidance from the professor during the discussion.
Interview with Processor Wang provided us with valuable insights that shifted the direction of our project somewhat. Initially, we focused on the role of chitinase in the purification of the marine environment, assuming that it could be effective in mitigating marine pollution. However, discussions with experts challenged this assumption.
An important revelation was that chitinase has a very specific function - it primarily breaks down chitin substances but is less effective in addressing broader marine pollution issues such as nutrient overload, inorganic pollutants, and plastic litter. This realization led us to reconsider the feasibility and impact of our initial research focus.
3.1.3 Reflection
Instead of focusing solely on the purifying effects of chitinase on the oceans, we recognized the potential for resource utilization of chitin. Discussions about the carbon sequestration properties of shellfish shells, their potential role in artificial coral reefs, and the possibility of extracting valuable materials such as chitin for industrial applications led us into a more practical and impactful direction of research. This is more in line with real-world environmental needs and economic viability.
Going forward, our program will move towards exploring the efficient enzymatic conversion of biomass to high-value resources. We will also conduct a comprehensive literature review to ensure scientific rigor and identify innovative approaches that balance environmental benefits with rational technology investments.
This expert meeting will be critical in refining our research focus, helping us to move away from impractical applications of chitinases to more viable and resource-efficient approaches to biomass conversion and marine environmental management.

3.2 Interview with Doctor Huang Limin
3.2.1 Expert Background
Dr. Huang Limin is responsible for the research and development of surgical suture materials, focusing on the synthesis and preparation of chitin and hyaluronic acid. Through interview, we have gained a new understanding of the contributions of chitin in the medical and corporate sectors, in addition to our previously set research directions in the food industry, agriculture, and fisheries.



Figure 29 Interview with Dr. Huang
3.2.2 Key Points
In terms of medical field, Professor Huang emphasized the impact of chitin on the destruction of bacterial cell walls and the activity of tumor molecules. By disrupting bacterial cell walls and altering their molecular structure, chitin can effectively aid in filling and antibacterial functions. Therefore, chitin can serve as a filler in surgeries and be synthesized into new chemical compounds for use as surgical adhesives. In terms of anti-tumor applications, similar to its effect on bacteria, injecting chitin to disrupt the cell walls of tumor molecules can effectively reduce their activity. Additionally, while cellulose is currently the largest category of polysaccharides on the market, its antibacterial effects are not as potent as those of chitin, giving chitin an advantage in medical applications. In terms of engineering applications, unlike substances such as acids and bases, chitin causes relatively less wear and tear on equipment and is more efficient and cost-effective. Regarding the economic feasibility of using chitin as a medical product, Professor Huang mentioned that the current procurement cost of pharmaceutical-grade pure chitin is high. If high purity and low cost can be achieved, the feasibility would be significantly higher. The temperature and pH ranges still need to be determined. If the conditions are too stringent, the feasibility would be lower, and it would not require specific environmental controls. As for breakthroughs in medicine, chitin has primarily been used in medical products like surgical sutures and some cosmetic products. However, there have been fewer breakthroughs in anti-tumor applications, as finding specific tumors and their corresponding effects takes time, and further research and clinical trials are needed. Moreover, Professor Huang also mentioned that chitin can be applied in lowering blood sugar and lubricating joints in the human body. Currently, small-molecule drugs are mainly used for lowering blood sugar, and while there are more biological drugs available, they are costly. If chitin is used to produce monosaccharide drugs, it would need to be mixed with biological agents, but the high cost remains a challenge. For joints lacking lubricating fluid, chitin can be formulated into long-chain polymers and injected into the needed areas, as it can degrade and thus alleviate the condition. From this perspective, the future demand for chitin is quite promising.
In the corporate sector, companies can effectively invest in the application of chitinase through mutual cooperation, thereby avoiding acquisitions. Various studies on chitinase need to be completed in the laboratory before moving on to corporate production and application. If specific diseases can be targeted, the development of drugs would be highly targeted and have good market prospects, though it requires multi-party collaboration.
3.2.3 Reflection
We have gained a deeper and more comprehensive understanding of the previously initiated research in agriculture and marine ecological protection. Chitin can help achieve carbon-nitrogen balance in the ocean, and its current use in marine pollution treatment is significant. Polysaccharides and monosaccharides, when dissolved in water, become viscous and form gel-like substances. If present in sufficient quantities, such as a layer on the sea surface, they may adsorb certain substances, but they do not contribute much to the treatment of algae or white pollution. The greatest contribution of chitinase is the degradation of shrimp and crab shells. Companies can collaborate with manufacturers, such as food processing plants and restaurants, to collect raw materials from source industries. The cost of pre-treatment is relatively low, as individual waste generation is minimal. Looking to the future, unless there is a breakthrough in research and development or existing methods overcome technical challenges, the application of chitin is unlikely to see significant breakthroughs.
3.3 Interview with Doctor Li Shiyuan
3.3.1 Expert Background
We have invited Dr. Li Shiyuan for an interview this time. Dr. Li Shiyuan graduated from the Institute of Plant Physiology and Ecology, Shanghai Academy of Sciences, Chinese Academy of Sciences, under the guidance of Academician Zhao Guoping, a leading figure in synthetic biology. My main research areas include heterologous synthesis of natural products, DNA splicing technology, DNA encryption, CRISPR gene editing, CRISPR detection (the inventor of CRISPR-Cas12), and many other fields. Our interview aims to understand the specific details of product pricing, address potential issues that may arise during the company's operations, and determine the significance of our project for sustainable development. We discussed the outline based on these objectives and hope to receive valuable guidance from the professor during the discussion process.

Figure 30 Interview with Dr. Li Shiyuan
3.3.2 Key Points
In our interview with Dr. Li Shiyuan, we gained valuable insights into the economic feasibility, industrial application, and commercialization of chitinase for the degradation of shrimp and crab shells. Dr. Li highlighted the importance of identifying target markets and potential customers, such as large food processing plants, pharmaceutical companies, and the biomedical sector. He also emphasized that while the environmental significance of our project is clear, we need to further analyze the economic sustainability of our approach.
From an industrial perspective, Dr. Li suggested that our initial focus should be on high-value applications such as medical, research, and cosmetic fields, where pricing power is stronger. Later, as production efficiency increases, we can expand to broader applications, such as agriculture and biofuel production.
Regarding pricing and market positioning, Dr. Li noted that our chitinase's ability to be reused gives it a competitive advantage, potentially justifying a higher price point. However, this would require detailed market research and comparison with existing solutions. He also pointed out the broader impact of synthetic biology in the pharmaceutical industry, where advancements in gene and cell therapies are creating new opportunities.
To scale up production, Dr. Li emphasized the need to differentiate our product in terms of application scenarios, optimize production costs, and consider supply chain logistics. Furthermore, he discussed different strategies for increasing profitability, including product diversification, cost reduction, and scaling effects.
Finally, we explored the sustainable development aspects of our project. Dr. Li encouraged us to consider alternative applications of chitinase, such as in agriculture or biodegradable pesticides, to further enhance our contribution to environmental sustainability.
3.3.3 Reflection
This interview provided us with a clearer understanding of the challenges and opportunities in bringing our chitinase-based technology to market. One of the key takeaways is the necessity of conducting in-depth market research to determine potential customers and pricing strategies. We also realized the importance of considering real-world industrial constraints, such as production scalability, regulatory hurdles, and market demand.
Dr. Li's insights into the synthetic biology industry's past successes and failures gave us a valuable perspective on how to approach our project's commercialization. The examples of successful companies like Novozyme and Cathay Biotech reinforced the importance of continuous innovation and adaptability.
Moving forward, we need to refine our business model, conduct feasibility studies for different applications, and explore potential industry partnerships. Additionally, we should consider the regulatory aspects if we aim to enter the pharmaceutical or food industries.
Overall, this interview was incredibly beneficial in shaping our strategic direction, reinforcing the importance of balancing scientific innovation with economic viability and sustainability.
3.4 Interview with Professor Han Zhenggang
3.4.1 Expert Background
A leading expert in synthetic biology and enzyme engineering, Professor Han's research focuses on enzyme optimization and industrial application, making him uniquely qualified to guide our technical development. His insights provided the theoretical and experimental framework to elevate our project from basic research to application-oriented innovation.

Figure 31 Interview with Professor Han
3.4.2 Key Points
Our initial experiments faced three critical bottlenecks:
1.Existing bacterial chitinases (from Streptomyces sp., Vibrio harveyi, and Serratia marcescens) showed limited efficiency in degrading complex chitin substrates (e.g., raw shrimp/crab shells).
2.Co-expressing multiple chitinase genes to enhance degradation required cumbersome vector construction, leading to low expression efficiency.
3.The cost of enzyme stabilization (via covalent crosslinking) was too high for large-scale use.
To address the core challenges of improving chitinase yield, activity, and industrial adaptability, we turned to Professor Han. He expertizes in enzyme source screening, expression system design, and cost-saving strategies directly targeted these issues, making him an indispensable advisor. Professor Han's guidance spanned degradation mechanisms, enzyme selection, and industrial optimization, each directly modifying our experimental protocol:
a.Enzyme Source: Prioritizing Fungal-Derived Chitinases
Professor Han noted that bacterial chitinases, while easy to express, often struggle with the complex, insoluble chitin in natural waste (e.g., shrimp shells with residual proteins and minerals). In contrast, fungal chitinases (e.g., from Aspergillus and Trichoderma) have evolved to degrade recalcitrant chitin structures due to their ecological role in decomposing fungal cell walls and insect exoskeletons.
b.Expression System: Simplifying Multi-Enzyme Co-Expression with Polycistronic Vectors
Our initial approach used separate plasmids for each chitinase gene, leading to uneven expression and low protein yields. Professor Han recommended polycistronic vectors, which allow multiple genes to be transcribed from a single promoter, ensuring balanced expression and reducing cloning complexity.
c.Cost Reduction: Chitosan Immobilization as a Replacement for Covalent Crosslinking
Covalent crosslinking (using glutaraldehyde) is a common method to stabilize enzymes but is expensive and toxic. Professor Han suggested chitosan immobilization—chitosan, a chitin derivative, is cheap, biodegradable, and forms stable complexes with enzymes via electrostatic interactions.
d.Substrate Pretreatment: Enhancing Accessibility with Steam Explosion
Raw chitin waste (e.g., crab shells) has a dense structure that limits enzyme access. Professor Han proposed steam explosion—subjecting substrates to high-pressure steam (1.5 MPa, 180°C) for 5 minutes—to break glycosidic bonds and expose more active sites.
3.4.3 Reflection
Professor Han's input shifted our technical focus from maximizing lab-scale activity to optimizing for industrial conditions. By prioritizing natural substrate compatibility, simplified expression, and cost efficiency, we laid the groundwork for a technically robust system that could transition from bench to pilot scale.
To enhance chitin degradation efficiency, we expanded our enzyme library by incorporating three fungal chitinases (GenBank IDs: XP_001219034.1 from Aspergillus niger, EGU72251.1 from Trichoderma reesei, and KAF3224578.1 from Penicillium oxalicum). Comparative assays revealed that the A. niger chitinase exhibited a 35% higher degradation efficiency on raw shrimp shells compared to our original bacterial enzymes. To improve expression efficiency, we designed a polycistronic vector under the E. coli T7 promoter, incorporating ribosome binding sites between the chitinase genes. This strategy reduced vector construction steps by 40% and increased total enzyme yield by 2.3-fold. For industrial application, we adopted chitosan beads for enzyme immobilization, which retained 80% activity after 10 reuse cycles—significantly higher than glutaraldehyde-crosslinked enzymes—while reducing material cost by 58% per gram of enzyme. Additionally, we integrated steam explosion pretreatment for crab shells into our workflow, which resulted in a 2.1-fold increase in water absorption and a 47% higher degradation rate, further boosting process efficiency.
3.5 Interview with Mr. George Ye
To transform technical and industry insights into a sustainable business, we consulted George Ye, a private equity investor with deep experience in tech commercialization, particularly in synthetic biology. His guidance focused on market positioning, investor appeal, and scaling strategies, ensuring our project could attract funding and achieve long-term growth.
3.5.1 Expert Background
Technical excellence alone does not guarantee commercial success. We needed to understand how investors evaluate biotech projects, identify key market drivers, and structure a scalable business model. George Ye's track record—he has funded 12 synthetic biology startups, 5 of which now generate more than ¥100 million in annual revenue—made him the ideal advisor to bridge science and finance.
3.5.2 Key Points
George Ye's advice centered on three pillars: validating market demand, building technical barriers, and planning for scalable production.
a.Minimum Viable Product (MVP) to Validate Market Fit
Investors prioritize tangible proof of market acceptance. Ye emphasized developing an MVP—a simplified version of our product—to gather real-world feedback and de-risk investment. For our project, he recommended a food preservation enzyme kit for small seafood processors, as it required minimal scaling and addressed a clear pain point.
b.Strengthening Technical Barriers to Prevent Competition
Ye warned that without patents, competitors could replicate our technology, eroding market share. He advised patenting not just the enzymes, but the entire workflow: polycistronic vector design, chitosan immobilization method, and steam explosion + enzyme degradation protocol.
c.Scalable Production: Leveraging Existing Infrastructure
Building dedicated fermentation facilities would require ¥50+ million in initial investment—prohibitive for a startup. Ye suggested partnering with contract manufacturing organizations (CMOs) that already have large-scale fermentation tanks (10,000L+), reducing capital expenditure by 80%.

Figure 32 Interview with George Ye
3.5.3 Reflection
To protect our core innovations, we filed three patents covering:
(1) A polycistronic vector for high-yield chitinase expression;
(2) A chitosan-based immobilization method for enzyme reuse;
(3) A combined steam explosion and enzymatic process for chitin waste degradation, establishing a strong technological barrier.
Meanwhile, we secured a manufacturing partnership with a food enzyme CMO in Jiangsu—equipped with 20,000L E. coli-compatible tanks—enabling scaled production at a cost of ¥42/kg, meeting targets for agricultural use. To validate market fit, we launched an MVP pilot with 5 local seafood processors in the form of 100ml chitinase preservative bottles. After three months, 4 clients renewed orders, reporting a 20% reduction in spoilage costs—a key outcome used in early-stage funding discussions.
George Ye's input transformed our business model from "technology-driven" to "market-driven". By focusing on an MVP, securing patents, and leveraging CMOs, we created a path to profitability within 3 years, making the project attractive to both investors and industry partners.
3.6 Visit to AstraZeneca
3.6.1 Company Background

AstraZeneca is a world-leading, innovation-driven biopharmaceutical company headquartered in London, UK, with operations in over 100 countries. It consistently stands at the pinnacle of the world's pharmaceutical industry, focusing on the research, development, and commercialization of drugs in three core therapeutic areas: Oncology; Cardiovascular, Renal & Metabolism; and Respiratory & Immunology. AstraZeneca's success stems not only from its powerful R&D capabilities but also from its adeptness at integrating cutting-edge technologies and embracing cross-disciplinary collaboration, consistently striving to translate scientific ideas into life-changing medicines.

Figure 33 Our Visit to AstraZeneca (Shanghai)
3.6.2 Key Points
During our visit to AstraZeneca, we focused on learning how it drives the healthcare industry forward through business innovation. AstraZeneca does not work alone. It actively collaborates with various partners, including governments, companies, universities, research institutions, hospitals, and investment firms. Their strategy can be summarized as building a Health Innovation Ecosystem, primarily achieved through three key initiatives:

Figure 34 Talking with AstraZeneca's Staff
a.China Centre for Health Innovation (CCiC)
The idea of this center is to start from patient needs and create innovative management models that cover the entire process from "screening, diagnosis, and treatment to management".

Figure 35 Listened to the Staff
It collaborates with partners from different fields to co-develop and implement integrated diagnostic and treatment solutions, driving the upgrade of the domestic medical industry. Another key task is to facilitate these successful Chinese innovations and models to expand abroad, allowing patients in other parts of the world to benefit.
b.International Life Science Innovation Campus (iCampus)
This is an incubation base built by AstraZeneca in collaboration with local governments, providing an open and international environment for healthcare innovation companies to grow.

Figure 36 iCampus
It offers comprehensive support, including organizing industry conferences, forming industry alliances, introducing high-quality projects, providing investment and incubation, and helping with the transformation of scientific research into practical applications.
c.AstraZeneca-CICC Capital Healthcare Industry Fund
This fund was jointly established by AstraZeneca and CICC Capital, specializing in investments in the healthcare industry. It combines AstraZeneca's global industry resources with CICC Capital's investment expertise. Key investment areas include biopharmaceuticals, medical devices, diagnostic services, and digital health.

Figure 37 Learned about AstraZeneca's Healthcare Industry Fund
The purpose of the fund is not only to provide capital but, more importantly, to offer all-around support for the companies it invests in—from product development to commercial success—thereby accelerating innovation in China's healthcare sector.
3.6.3 Reflection
Our visit to AstraZeneca provided new direction for our research project on chitinase. Previously, our focus was primarily on technical aspects in the laboratory. However, this visit broadened our perspective and showed us a bigger picture.
AstraZeneca does not work alone but partners with various organizations. This made us realize that our research could also benefit from seeking collaborations, such as with pharmaceutical companies or environmental agencies, to maximize the impact of our findings. We understood that scientific research should emphasize practical applications. AstraZeneca consistently focuses on how innovation can genuinely help patients. This reminded us to consider how our chitinase research can address real-world problems, such as developing new medicines or improving production processes.
Innovation requires resource support. Just as AstraZeneca has its industry fund to support innovative projects, good ideas need funding and resources to become reality. This encouraged us to think about how to refine our project, making it not just an experiment but a feasible idea with potential for real-world impact.
This visit taught us to look beyond the lab and consider our research from multiple perspectives—collaboration, practicality, and resource support—which will greatly inspire our future work.
3.7 Visit to Biotechnology Company
3.7.1 Company Background
To understand the practical challenges of applying chitinase in real-world scenarios, we partnered with Shanghai Plant Science Biotechnology Co., Ltd. (hereafter "Plant Science"), a leader in biopesticide development with over 10 years of experience in agricultural technology. Their expertise in commercializing biological products revealed critical gaps between our lab results and industrial requirements, guiding us to refine our application strategy. Our project initially targeted agricultural biopesticides, as chitinase's ability to degrade fungal cell walls makes it a promising eco-friendly alternative to chemical pesticides. However, we lacked insights into farmer needs, field conditions, and industry workflows. Plant Science, which specializes in RNA-based biopesticides, provided a frontline perspective on the barriers to adopting new biological products.

Figure 38 Our Trip to Plant Science
3.7.2 Key Points
Plant Science's technical team, led by their R&D director, highlighted three critical challenges we had overlooked:
a.Field Stability: Enzyme Sensitivity to Environmental Factors
Lab tests showed our chitinase was active at 37°C and pH 7, but field conditions are far more variable. The team noted that in agricultural settings, enzymes face UV radiation (which denatures proteins), fluctuating pH (5.0–8.5 in soil), and temperature extremes (10–40°C). Their data showed even stable enzymes lose 60–70% activity within 48 hours in open fields.
Implications: Our current enzyme formulation was unsuitable for direct agricultural use. Short-term, we needed to target applications with milder conditions; long-term, we required stabilization technologies (e.g., microencapsulation) for field use.
b.Cost Constraints for Farmers
Plant Science's market research showed farmers in China (our primary target) are highly price-sensitive, with biopesticides needing to cost <¥50/kg to compete with chemical alternatives. Our lab-scale production cost (¥120/kg) was far too high, even with chitosan immobilization.
Implications: We needed to identify high-value, low-volume applications to justify higher initial costs while optimizing production to reduce long-term prices.
c.Integration with Existing Pest Management Workflows
Farmers rarely adopt standalone products; they prefer formulations compatible with existing practices (e.g., mixable with fertilizers or other biopesticides). Our enzyme's liquid form was unstable in mixes containing metal ions (common in fertilizers), limiting its practicality.
Implications: Formulation adjustment (e.g., powderization) and compatibility testing were necessary for widespread adoption.

Figure 39 Introducing us to the Core Advantages
3.7.3 Reflection
Plant Science's insights forced us to ground our innovation in industrial reality. By prioritizing food preservation as a stepping stone, we gained a path to market validation while continuing to develop agricultural applications—ensuring our project progresses iteratively rather than chasing unachievable short-term goals.

Figure 40 We presented Small Gifts to Staff
Guided by Plant Science, we revised our application strategy to balance ambition with feasibility:
a.Short-Term: Food Preservation
Food processing (e.g., seafood preservation) has milder conditions (controlled temperature, pH 6–7) and higher profit margins, making it a suitable first market. Plant Science noted that seafood processors lose ¥3 billion annually to spoilage, creating demand for natural preservatives.
We developed a chitinase-based preservative (1% enzyme solution) for shrimp. Trials with a local seafood company showed it extended shelf life by 3 days at 4°C (vs. 1 day for untreated samples) and met food safety standards (no toxic residues), validating its potential.
b.Long-Term: Agricultural Biopesticides with Stabilized Formulations
To address field stability, we collaborated with Plant Science to test microencapsulation—coating enzymes in alginate beads (5μm diameter) to block UV and buffer pH. Lab tests showed encapsulated chitinase retained 75% activity after 72 hours of UV exposure (vs. 15% for uncoated enzyme), bringing field application closer to reality.
3.8 Integrated Logic: From Insights to Iteration
The interactions with Professor Han, Plant Science, and George Ye form a closed loop of improvement, where each stakeholder's feedback addresses gaps revealed by the others:
3.8.1 Technical Feasibility → Industrial Applicability
Professor Han's optimizations (fungal enzymes, steam explosion) laid the groundwork for processing real-world waste, but Plant Science's input revealed that field conditions required further stabilization—prompting us to develop microencapsulation, using our chitosan expertise from Han's advice.
3.8.2 Industrial Applicability → Market Viability
Plant Science identified food preservation as a short-term market, which George Ye then refined into an MVP strategy, using the cost-saving immobilization method (from Han) to meet price targets.
3.8.3 Market Viability → Technical Refinement
Feedback from MVP trials (e.g., demand for longer shelf life) led us back to Professor Han for advice on enzyme stabilization, resulting in a modified chitosan formulation with 12-month stability (vs. 6 months initially).
This iterative cycle—technical optimization informed by industry needs, commercial strategy shaped by technical limits, and real-world feedback driving further innovation—defines our Integrated Human Practices. By centering stakeholder engagement at every stage, we ensure our chitinase project is not just scientifically sound, but also practical, scalable, and aligned with the needs of farmers, food processors, and the environment.
In the end, synthetic biology's greatest impact comes not from lab results alone, but from solutions that resonate with the world they aim to improve. Our IHP journey has ensured that our chitinase technology is one such solution.
3.9 Conclusion
The journey of our chitinase project—from technical design to practical application and commercial planning—has been guided by targeted engagement with several key stakeholder groups: academic experts, industry practitioners, and investment professionals. Each interaction addressed critical gaps in our knowledge, driving iterative improvements that transformed our initial concept into a solution rooted in real-world feasibility.

4.1 Who are your proposed end users?
Our proposed end users primarily include seafood processing plants, agricultural sectors, and biotechnological or pharmaceutical companies. Seafood processors can utilize our chitinase-based preservatives to extend the shelf life of products such as shrimp, reducing spoilage losses. Farmers and agricultural cooperatives may adopt our enzyme formulations as eco-friendly biopesticides to combat fungal pathogens. Additionally, pharmaceutical and cosmetic industries could leverage high-purity chitin derivatives for applications in wound healing, drug delivery, or functional biomaterials.

4.2 How do you envision others using your project?
We envision others using our project through accessible, scalable enzyme products tailored to specific applications. For instance, food processors may apply our ready-to-use chitinase solution as a natural preservative spray or dip. Agricultural users could incorporate powdered or microencapsulated chitinase formulations into existing integrated pest management systems. Researchers and industrial partners might also employ our patented polycistronic expression system and immobilization techniques to produce chitinase variants for customized applications.
4.3 How would you implement your project in the real world?
To implement our project in the real world, we will adopt a phased approach: First, we will partner with contract manufacturing organizations (CMOs) to scale enzyme production using large-scale fermentation infrastructure, ensuring cost-effectiveness and consistent quality. Next, we will validate market fit through pilot programs with seafood processors and agricultural cooperatives, gathering feedback to refine product formulations. Simultaneously, we will pursue regulatory approvals and intellectual property protection to secure commercial viability. Finally, we will expand into high-value markets such as pharmaceuticals by collaborating with industry partners to develop certified chitin-derived products, ensuring our technology delivers both environmental and economic benefits across multiple sectors.