Figure 2 *Have you ever used antibiotics before?

Apparently, most people have ever used antibiotics like Penicillins and Cephalosporins, accounting for nearly 90%, which means antibiotics are really common in humans’ daily life.

Figure 3 *Are you aware that antibiotic overuse can lead to bacterial antibiotic resistance?

Besides, about 82% of respondents have heard about the risk of inappropriate antibiotic use. This indicates that the population has a generally good understanding of antibiotics.

Figure 4 *Have you previously considered the risks of antibiotics, leading you to seek antibiotic alternatives?

While the percentage of respondents who want to mitigate antibiotic resistance by finding an alternative is not as high as that in the previous question, which is 50.58%. This demonstrates that the collective perception currently only scratches the surface of issue.

Figure 5 *Have you ever heard of AMP before conducting the questionnaire?

Then we introduce AMP(antimicrobial peptides) and their function to our participants. As expected, 90.41% of respondents don’t know about this technical term (AMP).

Figure 6 *After reviewing the potential drawbacks of antimicrobial peptides (AMPs) mentioned above, would you be willing to use AMPs as an antibiotic alternative?

Additionally, a small number of respondents are willing to take AMP as an alternative, and 77% of them are either unsure or simply disagree. In other words, their replies to this novel approach are understandably noncommittal. The consequences reflect that more information and science outreach programs are needed to support for the population in order to help them make a decision.

Figure 7 *Through which channels would you prefer to obtain information about antimicrobial peptides (AMPs)? (Multiple selections allowed)

Concerning the way of getting the information about AMP, respondents’ preferences are evenly distributed. More than half of the people want to learn about AMP through survey reports. As a result, a large number of people choose literature and studies as the source due to the high credibility of such materials. Also, lectures and short videos are well-received.

Figure 8 *If antimicrobial peptide (AMP) products were commercially available, what price premium over conventional alternatives would you find acceptable?

Originating from the characteristics of AMP, we made an inquiry about the price they expect. According to the questionnaire, it is clear that most of the respondents accept the less than 10% of price premium, indicating that they are sensitive to price change. So, they tend to purchase products with a high price-performance ratio.

Figure 9 *What characteristics do you consider essential for an ideal antimicrobial peptide (AMP) drug in the future? (Multiple selections allowed)

This finding suggests that pricing requires careful consideration. What’s more, we found that respondents consider a high level of safety and fewer side effects as essential features for an ideal AMP drug.

In conclusion, our survey revealed a clear gap: while the public is aware of the risks of antibiotic resistance, few understand what AMPs are or how they can help. This finding guided us to design education strategies that not only raise awareness of AMPs but also build trust in their safety and effectiveness. It also shaped our project planning—ensuring that communication runs in parallel with technical development, so that when AMP-based solutions reach the market, consumers are prepared to accept them.

The specialized version of the questionnaire mainly targeted professionals engaged in related scientific research, aiming to understand their views on antibiotics and antimicrobial peptides (AMPs).

Figure 10 *What is your current field of research?

The respondents came from various fields, including biochemistry and molecular biology, medical devices, and biopharmaceuticals. Most of them were biochemical researchers (38.1%) or from other related professions (42.86%).

Figure 11 *Have you ever approached Pichia Pastoris

However, the survey results show that only a small proportion (38.1%) of respondents have experience working with Pichia pastoris, indicating that this area remains relatively underdeveloped.

Figure 12 *What do you think the most promising application scenarios for AMP? (Multiple selections allowed)

Regarding the application scenarios for AMPs, participants showed interest in fields such as biopharmaceuticals, preservatives, medical sterilization, and replacing antibiotics in animal husbandry, with no significant differences among the choices (all selected by more than half). Among them, biopharmaceuticals and antibiotic alternatives in animal husbandry were regarded as having the greatest potential (both at 66.67%).

Figure 13 *What do you think the most advantage of AMP currently？(Multiple selections allowed)

As for the greatest advantage of AMPs, the enhancement of the host’s natural defense system ranks first among the options (71.43%), indicating that respondents recognize AMPs' ability to boost the host’s resistance to pathogens. This is followed by broad-spectrum antimicrobial activity (57.14%) and low risk of resistance development (52.38%).

Figure 14 *What do you think is the biggest challenge in the expression of AMP in yeast? (Multiple selections allowed)

Most respondents (57.14%) believed that susceptibility to protease degradation and the complexity of production processes are the biggest challenges for AMPs expressed in yeast. This reflects that proteases significantly impact the stability of AMPs, while the production process can also lead to their loss.

Figure 15 *What do you consider a reasonable post-production price for antimicrobial peptides (AMPs)?

The majority(57.14%) still believed that the price of AMPs should remain below 10 yuan per gram, reflecting the importance of price as a factor, which aligns with public expectations. This suggests that people hope the price of AMPs will stay within a reasonable range to enable widespread adoption. This result is likely influenced by the fact that current antibiotic market prices are relatively low.

Figure 16 *Do you believe gene-editing technologies (e.g., CRISPR) could enhance antimicrobial peptide (AMP) production?

The vast majority agreed that gene editing technologies such as CRISPR can effectively improve AMP yields, reflecting broad recognition and trust in such technologies. At the same time, the public also supports the application of innovative technologies to enhance products and improve human well-being.

Figure 17 *How widely do you anticipate antimicrobial peptides (AMPs) will be adopted in the market over the next 5-10 years?

Most participants believe that AMPs will become widely used in the future, with approximately 42% believing they will become mainstream antimicrobial products. This reflects an optimistic attitude towards the future development of AMPs.

The survey yielded valuable insights into the perceived advantages and practical challenges of antimicrobial peptides (AMPs). Respondents noted AMPs’ strengths—broad-spectrum activity, enhanced host defense, and low resistance risk—underscoring their potential in biopharmaceuticals and as antibiotic alternatives in animal husbandry. It also identified key hurdles: protease susceptibility, complex production, and the need for cost-effective post-production pricing.

These findings directly shaped the project: recognition of CRISPR-like gene-editing technologies reinforced the exploration of synthetic biology to boost AMP yields; concerns over production complexity and pricing guided prioritization of scalable, cost-conscious designs; and the data helped tailor outreach education (highlighting both AMPs’ scientific potential and practical limits) to improve public understanding.

Overall, the survey embodies mutual learning: expert opinions refined the project, while the feedback loop prompted reflection on communication strategies, adaptation of educational materials, and planning of future research. Integrating these insights across modules (lab design to community education) ensures the AMP work is scientifically sound, socially responsible, and accessible to wider audiences.

Figure 18 Insights from interview with Prof. HUO

To refine our synthetic biology project on antimicrobial peptides (AMPs), we interviewed Professor Huo, an expert in microbial expression systems and peptide engineering. The goal was to obtain professional feedback on current expression platforms—especially Pichia pastoris—and to better understand the production challenges and optimization strategies for AMPs such as Mycosin.

Professor Huo offered deep insights into both chemical and biological synthesis of AMPs. He explained that while chemical synthesis is suitable for short, linear peptides, its cost increases sharply with structural complexity or longer sequences. In contrast, biological synthesis—especially via microbial hosts—is more scalable and cost-effective for complex or longer peptides. However, challenges remain: chemical methods struggle with yield and purity, while biological methods face difficulties in secretion, stability, and purification.

He emphasized that key factors such as expression efficiency, product stability, and purification complexity are central to cost and scalability. Notably, AMPs are inherently antimicrobial, often exerting toxicity toward host cells, which reduces expression yields. Additionally, their strong hydrophobicity, charge distribution, and sensitivity to environmental conditions complicate purification and storage.

To improve expression, Professor Huo recommended several strategies. First, using high-strength promoters and codon optimization can significantly enhance transcription and translation. However, he noted that without efficient secretion pathways, intracellularly accumulated peptides are prone to degradation. Thus, expression strength and secretion efficiency must be co-optimized. He also highlighted the utility of CRISPR technology in fine-tuning gene elements, such as inserting strong promoters or adding purification tags to the N-terminal of the peptide.

Beyond technical guidance, Professor Huo offered advice on application pathways. He pointed out that AMPs act by disrupting bacterial membranes, differing from conventional antibiotics that often inhibit biosynthetic pathways. This unique mechanism gives AMPs lower toxicity and broader antimicrobial potential. While regulatory hurdles remain high for clinical use, AMPs show strong potential in non-therapeutic applications, such as food preservation and animal feed additives, where safety requirements are less stringent.

Importantly, Professor Huo encouraged us to stay focused on our project’s core goal—efficient expression of Mycosin—while choosing technologies that directly support this aim. He also suggested combining biological and chemical synthesis approaches to balance cost, complexity, and purity for practical deployment.

This interview gave us a clearer direction for optimizing our expression system. Moving forward, we will incorporate Professor Huo’s feedback by enhancing our vector design, exploring secretion pathways, and carefully evaluating application scenarios to develop a scalable and market-ready AMP product.

Figure 19 interviewed with Prof. Huo

Figure 20 Insights from interview with DR. Cumbers

To explore the commercial potential and communication strategy for our antimicrobial peptide (AMP) project, we interviewed Dr. Cumbers, founder and CEO of SynBioBeta—the world’s leading synthetic biology community focused on promoting innovation for a more sustainable future. A former iGEM Ambassador who played a crucial role in introducing iGEM to China, he is also an experienced biotech investor and a strong advocate for public science communication. The goal of this interview was to gain insights into AMP commercialization, stakeholder engagement, and science outreach from a global leader in the synthetic biology field.

Dr. Cumbers expressed strong interest in our project and emphasized that synthetic biology holds great promise in addressing antibiotic resistance. After learning about our approach—using promoter engineering and a high-efficiency *Pichia pastoris* system to enhance AMP production—he affirmed that such metabolic optimization is essential for real-world applications. He encouraged us to continue improving yield and scalability to ensure downstream viability.

Importantly, Dr. Cumbers stressed that scientific innovation alone is not enough. A successful synthetic biology project must also identify its early customers and communicate value clearly. “You need to think about how to make money,” he said, reminding us that bridging biology and business is crucial. He advised targeting specific sectors—such as governments, clinics, or farms—with both demand and budget, while ensuring the production model allows for sustainable profit.

On the topic of commercialization, Dr. Cumbers pointed out that challenges go beyond lab-scale success. Teams must navigate regulatory frameworks, define the product’s identity (e.g., food additive, veterinary drug, or therapeutic), and build trust through user education and consistency in quality.

As a long-time advocate for science outreach, Dr. Cumbers also emphasized the importance of public communication. “People don’t trust what they don’t understand,” he noted, encouraging us to make synthetic biology approachable through storytelling and visual media. He highlighted short-form videos as especially powerful for reaching younger audiences, referencing his own recent participation in an iGEM promotional shoot in Tianjin.

This interview provided valuable guidance on aligning our technical innovation with market and communication strategies. Moving forward, we will integrate Dr. Cumbers’ advice to refine our commercialization plan, proactively engage key stakeholders early, and use creative media to expand public understanding and support for synthetic biology.

Figure 21 interviewed with Dr. John Cumbers

Figure 22 photo with Dr. John Cumbers

Figure 23 Insights from interview with Dr. Wang

To explore the clinical relevance and application potential of antimicrobial peptides (AMPs), especially Mycosin, we interviewed Dr. Wang, a medical doctor and lecturer at Wuhan University, who also serves as Associate Chief Physician in the Physical Examination Department at Wuhan Petrochemical Hospital and practices in the Emergency Department at Wuhan University People’s Hospital. The goal of this interview was to understand current challenges in antibiotic use and evaluate the clinical feasibility of AMPs as alternative therapeutics.

Dr. Wang confirmed that antibiotic resistance is a frequent and growing issue in clinical practice, particularly in ICU and emergency departments. He noted that infections caused by multidrug-resistant bacteria such as MRSA, CRE, and Acinetobacter baumannii significantly complicate treatment and increase mortality. He acknowledged that AMPs, with their broad-spectrum activity and unique membrane-disrupting mechanisms, show promising potential as low-toxicity alternatives, especially in cases where conventional antibiotics fail.

While strict prescription protocols exist in his hospital to control antibiotic use, Dr. Wang emphasized that resistance persists due to long-term misuse in both human medicine and agriculture. This underscores the urgent need for novel antimicrobial agents.

Regarding clinical translation, Dr. Wang identified large-scale production as the primary barrier to AMP application. Natural extraction is inefficient and costly, making gene-engineered expression in eukaryotic systems—such as Pichia pastoris—a necessary direction for future development. He noted that yeast-based systems offer advantages in secretion, safety, and scalability.

Dr. Wang also suggested potential AMP application scenarios, including burn wound care (topical gels), ICU respiratory infections (IV formulations), catheter-associated UTIs, and post-surgical infection prevention (spray-based use). He added that AMPs may be especially suitable for vulnerable groups like children, pregnant women, and immunocompromised patients, for whom conventional antibiotics pose greater risks.

Finally, Dr. Wang noted that beyond production and cost, clinical guidelines and physician training will be necessary for broader adoption. However, these challenges are surmountable if AMPs can demonstrate reliable efficacy and safety in future trials.

This interview provided important clinical validation for our research direction and highlighted key opportunities and barriers in the clinical translation of AMPs. We will incorporate Dr. Wang’s insights into our design of application scenarios and scale-up strategies.

Figure 24 interviewed with Dr. Wang

Figure 25 Insights from interview with Mr. Wang and Mr. Liu

To understand the practical needs and adoption potential of antimicrobial peptides (AMPs) in dairy farming, we interviewed 2 experts: Mr. Wang, former Animal Welfare Manager at Yili Group and current Deputy Technical Director at Qianjin Modern Agriculture Group, and Mr. Liu, Deputy Ranch Director at Gansu Delian Animal Husbandry Co., Ltd. Both experts have extensive experience in large-scale cattle health management and provided key insights into antibiotic use, resistance trends, and the requirements for introducing AMP alternatives on farms.

Mr. Wang explained that their farms currently use antibiotics such as β-lactams and macrolides, administered mainly via spinal or intravenous injection. Resistance has become a significant challenge, especially in newly calved cows. He noted that repeated use of the same drug can lead to sharply reduced cure rates, prompting the farm to implement a tiered prescription protocol for treating mastitis and other infections.

Both experts agreed that while AMPs show promise as antibiotic alternatives, cost remains the deciding factor. They emphasized that farms do not only consider the unit price of a drug but its total treatment cost—including cure rate, labor, and milk discard losses. For example, with milk priced at ¥3/kg and an average daily output of 25 kg per cow, even three days of discarded milk due to antibiotic use leads to ¥225 in economic loss. A new product that shortens recovery time or avoids discard entirely would be highly attractive, even at a higher price point.

Regarding delivery methods, they noted that individual injection is preferred for isolated infections, while mass administration is unnecessary in the absence of widespread symptoms. For preventive or mild cases, their farms are already experimenting with non-antibiotic approaches such as anti-inflammatory therapy and herbal formulations, provided they are simple to use and cost-effective.

With increasing global restrictions on antibiotic residues, especially in export-oriented markets like the EU and South Korea, both experts believe that antibiotic-free solutions will become a long-term requirement. Their companies are actively evaluating alternatives and welcome AMP-based innovations—if they are well-targeted, easy to administer, and supported by reliable field data.

This interview offered important operational and economic perspectives on AMP adoption. Based on these insights, we will refine our product strategy to align with real-world farm practices and ensure our solution meets industry needs in both efficacy and practicality.

Figure 26 Interview with Mr. Wang and Mr. Liu

Figure 27 Insights from Field Trip to Zoetis

Zoetis, a global leader in animal health, originated as the animal health division of Pfizer and became an independent company listed on the New York Stock Exchange in 2013. It is dedicated to delivering innovative medicines, vaccines, diagnostic tools, and digital solutions for both companion animals and livestock. Today, Zoetis operates in nearly 100 countries and regions and continues to expand its presence in the Chinese market by investing in localized research and production.

The primary objective of this field visit was to conduct an in-depth interview with Dr. Wu, Senior Technical Services Director for Strategic Clients in the Swine Business Unit of Zoetis China. The discussion focused on three key areas: the current use of antibiotics in pig farming, challenges related to antimicrobial resistance (AMR), and the potential of antimicrobial peptides (AMPs) as alternative solutions in animal health.

Figure 28 Photo in Zoetis

Expert Insights and Evaluation

Dr. Wu provided highly constructive feedback on our project:

He acknowledged the enormous potential of AMPs as future mainstream alternatives to antibiotics in both animal husbandry and human medicine.

He highlighted the inherent drawbacks of traditional antibiotics, such as resistance and residue risks, and shared that Zoetis has already reduced investment in further antibiotic R&D to focus on new-generation antimicrobials.

He endorsed the use of AMPs in livestock, emphasizing their relatively low risk for food safety concerns, especially after cooking and processing.

He noted the market gap for high-quality and high-yield AMP products, validating the innovative value of our project.

Suggestions and Strategic Recommendations

Dr. Wu also offered practical guidance for improving and positioning our product:

Strategic Market Reflection

This visit highlighted that while AMPs hold great promise, market adoption depends on a balance between innovation, scientific rigor, economic feasibility, and operational simplicity. From production to dosage and administration, every step must be aligned with the realities of livestock management and food safety expectations.

Our exchange with Zoetis provided not only a deeper understanding of industry needs but also clear directions for refining our product. The feedback will guide us as we continue to develop a reliable, cost-effective, and scalable AMP-based solution for the animal health industry.

Figure 29&30 Interview with Dr. Wu