Lactose intolerance is a widespread health condition characterized by the body's inability to effectively digest lactose—the primary sugar in milk and dairy products—due to insufficient lactase enzyme production. This condition is prevalent worldwide, yet its distribution exhibits notable ethnic and geographical variations. For instance, across much of Africa, the prevalence of lactose intolerance ranges between 65% and 75%. In East and Southeast Asia, the figures are even more striking, with over 90% of the adult population affected to varying degrees. This means that, globally, hundreds of millions of people face challenges related to lactose malabsorption.

The condition substantially impacts patients' daily lives. When lactose is not broken down, it ferments in the colon due to bacterial action, leading to a range of gastrointestinal symptoms such as bloating, abdominal pain, flatulence, diarrhea, and nausea. Beyond physical discomfort, these symptoms often compel affected individuals to long-term avoidance of dairy products. Such dietary restrictions not only diminish their quality of life but may also result in insufficient intake of essential nutrients like calcium and vitamin D, thereby increasing the risk of osteoporosis and other metabolic disorders.

Currently, both diagnosis and management of lactose intolerance are hampered by systematic shortcomings in clinical practice, leaving patients without fully satisfactory solutions.

On the diagnostic front, although widely used methods exist, their application experience and accuracy face significant bottlenecks. The hydrogen breath test is recognized as the gold standard for diagnosing lactose intolerance due to its high sensitivity and specificity. However, the testing process is cumbersome, time-consuming (lasting 2-3 hours), and requires strict patient adherence to pre-test preparations (e.g., discontinuing antibiotics weeks in advance, following specific diets, and avoiding smoking or exercise on the test day). More importantly, the results are prone to false negatives (e.g., due to gut flora that do not produce hydrogen) or false positives (e.g., due to small intestinal bacterial overgrowth), which can compromise diagnostic accuracy. Genotyping via oral mucosal cell or blood testing (e.g., for the C/T-13910 SNP) serves as an effective alternative for distinguishing primary from secondary lactase deficiency. However, its applicability is limited in non-Caucasian populations, and it cannot reflect the real-time severity of symptoms. The complexity of diagnosis leaves a large number of patients undiagnosed or misdiagnosed for extended periods.

In terms of treatment and management, existing approaches remain largely reactive and fail to address the root cause.

Dietary Avoidance: As a passive management strategy, it severely restricts patients' dietary freedom, impairs social interactions and quality of life, and carries potential long-term nutritional health risks.

Oral Lactase Supplements: This is currently the primary remedial measure, but its effectiveness is highly unpredictable. The activity of exogenous lactase tablets is easily compromised by stomach acid, and matching their timing, optimal dosage, and meal composition is challenging, leading to inconsistent and unreliable results. In related interviews and surveys, patients often report symptom recurrence even after supplementation.

Traditional Probiotic Products: While some probiotics are marketed as aiding lactose metabolism, these strains generally exhibit weak colonization ability and low survival rates in the gut. Their effects are mild, slow-acting, and subject to significant individual variation, failing to provide consistent and noticeable symptom relief.

Initially, we will focus on the Mongolian community in China's Inner Mongolia region as our beachhead market. This group aligns exceptionally well with our solution:

High Prevalence and Cultural Conflict: Despite the traditional reliance on dairy products in Mongolian culture, modern genetic studies indicate a considerable prevalence of lactose intolerance. This forces many individuals to endure physical discomfort when consuming millennia-old traditional foods like milk tea and dried milk curds, creating a direct conflict between cultural practices and personal health.

Urgent Need for Cultural Preservation: Milk tea (süütei tsai) holds profound significance in Mongolian culture—it is central to hospitality, a staple of social and ceremonial gatherings, and carries deep historical and emotional value. Our solution enables them to continue this precious tradition without physical burden, positioning it not merely as a health product but also as a tool for cultural preservation. This carries strong emotional appeal and market acceptability.

Ideal Validation Scenario: This community possesses a strong cultural identity and is relatively concentrated geographically, facilitating initial market education, clinical observation, and user feedback collection. It serves as an ideal setting for validating product efficacy and building early word-of-mouth.

Upon successful validation and refinement of our product and service model, we plan to expand across China and East Asia, initially targeting regions with long histories of dairy consumption and high lactose intolerance prevalence (e.g., Xinjiang, Tibet). Ultimately, our goal is to serve consumers worldwide who seek to break free from the constraints of lactose intolerance and pursue unrestricted dietary freedom.

It is foreseeable that our startup idea will encounter significant difficulties during its implementation. Therefore, we employ Porter's Five Forces model to analyze the potential challenges:

Competitive Rivalry Within the Industry We believe this will be the most immediate and intense level of competition faced after the project's commercialization, primarily stemming from established products and brands that already occupy consumer mindshare.

Major Competitor Categories & Representatives:

Oral Lactase Supplement Giants: e.g., Lactaid (global brand), Yili Lactic Acid-Free Milk (local benchmark). They define the current market's "standard therapy," leveraging strong brand power and channel penetration to ingrain a "pre-meal remedy" consumption habit among consumers.

Traditional Probiotic & Digestive Health Brands: e.g., Culturelle, Yakult. They have built deep trust in the "gut health" domain and could potentially create market confusion and interception by launching so-called "high-efficiency lactose-degrading strains."

Lactose-Free/Plant-Based Food & Beverage Giants: e.g., Danone (Silk brand), Oatly, Yili/Mengniu Lactose-Free product lines. Their business models are built on consumer demand for "avoiding lactose," and they will reinforce their products' positioning as "safe, fashionable alternatives" through lifestyle marketing.

Key Points of Competition: Fundamental efficacy (therapeutic) vs. immediacy (remedial); disruptive product innovation (live biotherapeutic) vs. established safety (dietary supplement); and shifting consumer usage habits.

Threat of New Entrants Once the project gains some attention and validates the value of this market segment, the entry of well-resourced "industry giants" will be attracted, posing the most significant long-term strategic threat.

Major Potential Entrants & Representatives:

Large Multinational Pharmaceutical Companies: e.g., Johnson & Johnson, Pfizer. They possess unparalleled financial resources, top-tier R&D platforms, and mature global clinical registration and pharmaceutical sales networks, enabling rapid imitation or the pursuit of more disruptive research.

Biotech Companies Focused on the Microbiome: e.g., Seres Therapeutics. They are the most direct peers in terms of technological pathways, with extensive experience in strain design, formulation, and clinical development, making technological migration barriers relatively low.

Health Industry Companies with Strong Distribution Channels: e.g., Nestlé Health Science. They might adopt a "fast-follower" strategy, leveraging their vast distribution networks covering pharmacies, hospitals, and e-commerce to exert market pressure and potentially overtake later.

Key Points of Competition: Speed of R&D and market entry; strength and breadth of patent protection; and the competition for core talent.

Threat of Substitute Products These are solutions that fulfill the same need of "consuming dairy products freely" but are based on completely different technological principles, potentially entering the market from the same angle.

Major Substitute Categories & Assessment:

Fecal Microbiota Transplant (FMT): A theoretical "natural microbiome" alternative. However, it faces core shortcomings such as difficulties in standardization, potential safety risks, and low patient acceptance, making it unlikely to become an effective competitor in the short term.

Symptomatic Relief Medications: e.g., antidiarrheal drugs like Loperamide. They only alleviate symptoms without addressing the root cause. Nevertheless, for patients with mild or occasional symptoms, their "low cost and convenience" can sufficiently meet certain needs, thereby weakening the motivation to try a fundamental therapy.

Key Points of Competition: Thoroughness of efficacy, invasiveness and safety of the treatment process, and comprehensive cost comparison.

Bargaining Power of Suppliers After analyzing our project, we conclude that its success relies on highly specialized and scarce upstream resources, resulting in strong bargaining power for suppliers.

Key Supplier Groups & Sources of Bargaining Power:

Core Intellectual Property and Tool Providers: e.g., Broad Institute (holder of key CRISPR patents). They control critical, non-circumventable technology patents, granting them absolute leverage over licensing fees and terms.

Contract Development and Manufacturing Organizations (CDMOs): e.g., WuXi Biologics, Lonza. GMP-compliant manufacturing capacity for live biotherapeutic products is a scarce resource. Especially during the early stages of the project when production scale is small, our position is relatively weak in scheduling and price negotiations.

Top Scientific Talent: Leading scientists in the field of synthetic biology are a scarce resource, necessitating high compensation and equity incentives for attraction and retention, directly increasing R&D costs.

Management Focus: Mitigating this dependency should involve considering long-term strategic partnership agreements, proactively developing in-house production capabilities, and building a robust patent portfolio.

Bargaining Power of Buyers We believe the bargaining power of buyers will increase significantly as the project moves from the market introduction phase into the growth and maturity phases.

Buyer Groups & Evolution of Bargaining Power:

End Patients (Individual Consumers): During the initial product launch, early adopters with the most acute need are relatively price-insensitive. However, as the market expands, a broader consumer base will compare its cost-effectiveness against lactase supplements, gradually increasing their bargaining power.

Large Hospitals and Medical Insurance Institutions: e.g., National Healthcare Security Administration, various commercial insurance companies. This represents the greatest source of future pricing pressure. To achieve inclusion in reimbursement formularies, we must provide robust health economic evidence demonstrating its long-term value. These payers will exert significant pressure to reduce prices through models like "volume-based procurement" and "price negotiations."

Online Health Platforms: e.g., Alibaba Health. They control massive consumer traffic and may demand "exclusive distribution rights" and "high commission rates," impacting our channel profitability and pricing autonomy.

Management Focus: Building strong brand premium and physician/patient loyalty in the early stages, and preparing sufficient "value-based healthcare" evidence for future negotiations with payers.

A random sample of 500 individuals from Northeast China was collected to analyze their dietary habits, awareness of lactose intolerance, and potential symptoms related to lactose intolerance.

Within the surveyed sample, a significant 68.7% (343/500) of respondents reported experiencing at least one gastrointestinal symptom following the consumption of dairy products. The breakdown of these symptom types is illustrated in the accompanying figure.

Considering the potential correlation between dietary patterns and lactose intolerance manifestations, we hereby present the relevant dietary profile data from the surveyed cohort.

Notably, only 6.6% of the total sample had received a clinical diagnosis, revealing a substantial gap compared to the 68.7% symptom reporting rate. This indicates that a significant number of symptoms are either overlooked or misattributed by individuals.

Further analysis by age group shows that the 18-30 cohort demonstrates the highest self-awareness rate of lactose intolerance (45.1%) but the lowest clinical consultation rate (9.8%). In contrast, the 50+ group shows lower self-awareness (28.6%) but the highest consultation rate (25.0%).

Regarding self-perception of lactose intolerance, the distribution among the three options was: Yes 38.6%, No 35.4%, and Uncertain 26.0%.

A notable cognitive-symptom discrepancy emerges: compared to the 68.7% symptom reporting rate, only 38.6% self-identify as lactose intolerant. This suggests approximately 30% of symptomatic individuals do not associate their discomfort with lactose intolerance, indicating that public awareness of this condition lags significantly behind its actual prevalence.

The substantial "Uncertain" cohort, exceeding one-quarter of respondents, undoubtedly signals considerable market potential. Based on these findings, we have conducted a prioritized analysis of our target demographic and formulated corresponding strategies.

First Priority: High-Potential Undiagnosed Young Adults (18-30 years) This group represents the second-largest cohort in the sample. They exhibit the highest self-awareness rate (45.1%) but the lowest clinical consultation rate (9.8%). They understand the concept of "lactose intolerance" and can vaguely associate symptoms with dairy intake, but tend towards self-diagnosis and endurance, being reluctant or not perceiving the need for formal medical consultation.

Core Strategy: Digital Educational Marketing & Self-Diagnosis Tools.

Tactics:

Utilize short-form video and social media feed advertisements for precise targeting with content themed: "Bloating, stomach gurgling, diarrhea? You might have unrecognized lactose intolerance."

Develop online symptom self-assessment tools based on classic symptoms to guide users toward self-identification and directly link them to the product solution.

Second Priority: Middle-Aged Patients Seeking Fundamental Solutions (31-50 years) This is the largest cohort in the sample. They are at the peak of career and family pressures and have higher demands for quality of life and health efficiency. They have likely tried various methods (e.g., avoiding dairy, occasional enzyme supplements) with unsatisfactory results and are weary of solutions that "address symptoms, not the root cause."

Core Strategy: Value-Driven Marketing, emphasizing "Fundamental" benefits and "Efficiency."

Tactics:

Design family bundles: Considering the strong family orientation in this age group, launch a "Family Share Plan" to encourage purchases for similarly affected spouses or children, thereby increasing the average order value.

Highlight the product's value proposition of "One-time treatment, long-term freedom," positioning it as a health investment for enhancing long-term quality of life.

Third Priority: Symptomatic but Misattributing & Uncertain Individuals (All Ages) This includes the ~30% of symptomatic individuals lacking awareness and the 26% "Uncertain" group. They either attribute symptoms to a "sensitive stomach," "spoiled food," or simply don't understand what lactose intolerance is. The 50+ group is particularly representative, showing high consultation rates but low self-awareness, potentially leading to misdiagnosis with other gastrointestinal conditions.

Core Strategy: Public Science Education & Symptom Association Awareness.

Tactics:

Conduct public health education initiatives emphasizing that symptoms like bloating, borborygmi, and diarrhea occurring within a specific time window (30 minutes - 2 hours) after dairy consumption are typical indicators.

Collaborate with community clinics and physicians to incorporate simple lactose intolerance screening questionnaires into routine check-ups or consultations, helping these "hidden patients" identify the actual cause of their discomfort. You may view some of our attempts in educational outreach [here]

Through Ranch & Factory Market Research, we identified that certain animals can also exhibit lactose intolerance. This prompted us to explore whether our product could present opportunities for providing long-term solutions to lactose intolerance in animals.

Companion Animals The companion animal market (e.g., cats, dogs) demonstrates significant demand for lactose intolerance solutions. Although dairy products are not a primary food source, many pet owners frequently offer milk, cheese, etc., as treats. Research indicates that most adult cats and dogs experience varying degrees of lactose intolerance, with ingestion easily triggering gastrointestinal symptoms like diarrhea and vomiting. The current solution is limited to ceasing dairy product provision, lacking proactive intervention methods. Our developed lactase probiotic formulation could serve as a dedicated pet nutritional supplement, added directly to daily diet or drinking water, helping pets establish healthy lactose metabolism. This market is characterized by high payment willingness and rapid consumer decision-making. Collaboration with major pet food companies (e.g., Mars, Nestlé Purina) could integrate it as a premium functional ingredient into their product systems.

Livestock Industry In the livestock industry, lactose intolerance directly impacts economic efficiency. Particularly during the early life stage (e.g., calves, lambs), their underdeveloped digestive systems have limited capacity to digest lactose in milk replacers, often leading to nutritional diarrhea, which affects growth and development and can even cause mortality. Current solutions primarily rely on antibiotic control and nutritional formula adjustments, offering limited efficacy and potential antimicrobial resistance issues. Our product could function as a feed-grade probiotic additive, enhancing lactose metabolism in young animals, improving feed conversion rates, reducing diarrhea incidence, and promoting healthy growth. This application could involve collaborations with major agricultural conglomerates (e.g., CP Group, New Hope Group) to provide specialized farming solutions, creating significant economic value.

Following our market analysis, we present the TAM, SAM, and SOM metrics relevant to our industry, encompassing the majority of the lactose-free milk and lactase supplementation regimens.

TAM (Total Addressable Market): The total expenditure on managing lactose intolerance by all potential patients globally who could benefit from a fundamental therapeutic solution.

SAM (Serviceable Addressable Market): The subset of the TAM within the geographically targeted East Asian region – the total expenditure by lactose intolerance patients in this area who could benefit from a fundamental therapeutic solution.

SOM (Serviceable Obtainable Market): The portion of the SAM that the company can realistically capture in the short term, considering factors like stakeholder access and competitive landscape. In our case, this is initially China, the startup's launch country.

Following an in-depth investigation into existing diagnostic and treatment solutions for lactose intolerance and market demands, we have conducted a comprehensive Strengths, Weaknesses, Opportunities, and Threats (SWOT) analysis focused on our core solution—a genetically engineered probiotic live biotherapeutic product.

This analysis systematically outlines the fundamental advantages of our therapy compared to traditional options, highlighting its potential to address a significant unmet clinical need, while also objectively examining the challenges we may face in technology validation and commercialization. This SWOT analysis is crucial for defining the product's value proposition, formulating an effective clinical development strategy, and proactively planning risk mitigation measures during commercialization.

Strengths

Integrated Multi-Module Technology Platform: The project integrates lactase engineering, adhesin protein expression, and an AI-2 quorum sensing system, providing a customizable and re-programmable live bacterial chassis. This platform holds future potential for application in other intestinal diseases.

Fundamental Treatment Paradigm: Aims to rebuild the body's intrinsic lactose metabolism capability, offering a long-term or potentially permanent solution. This creates a generational advantage over existing "avoidance" and "remedy" products.

Built-In Intelligent Safety System: The ccdB toxic protein and AI-2 quorum sensing system form a self-regulating population control circuit, proactively preventing over-proliferation of the engineered bacteria. This design-integrated safety feature surpasses traditional genetic engineering approaches.

Production and Storage Convenience: The engineered probiotics can leverage established fermentation and lyophilization technologies. The final product is easy to store and transport, requiring no complex cold chain logistics, thereby reducing distribution costs.

Strong Patent Moat: Unique genetic circuit designs (particularly the quorum sensing-toxin protein circuit) and strain construction methods can form a tight intellectual property shield, creating high technological barriers.

Weaknesses

Complex Validation of In Vivo Colonization and Functional Stability: Validating the colonization efficiency of the engineered bacteria in the complex real-world human gut environment, the long-term functional stability of the genetic circuits, and their impact on the host's native microbiota requires expensive and protracted clinical studies.

Potential Risk of Horizontal Gene Transfer: Although the probability is low, the potential transfer of engineered plasmids to other gut bacteria remains a concern, despite safety designs to mitigate it. This is a focal point for regulatory scrutiny and public apprehension.

Long Clinical Development Cycle and High Cost: As a first-in-class live biotherapeutic product, it must follow a strict pharmaceutical regulatory pathway. The entire preclinical and clinical trial process could take many years and requires substantial funding.

Public Acceptance of Genetically Engineered Organisms: The "genetically modified" label may raise concerns and resistance among some consumers and markets, resulting in high market education costs.

Opportunities

Significant Unmet Clinical Need: Survey data indicates that up to 30% of symptomatic individuals are unaware of the cause, representing a vast, educatable, and convertible "silent market."

Strategic Partnerships with Dairy Giants: Potential collaboration with companies like Yili and Mengniu could convert their vast consumer base of "lactose-free" product users into our potential customers, or enable co-development of functional foods for mutual benefit.

Horizontal Expansion Potential of the Technology Platform: The validated chassis technology can be rapidly adapted to other fields, such as developing engineered bacteria for phenylalanine degradation in PKU or for oxalate degradation in hyperoxaluria, potentially multiplying the market scope.

Gradual Clarification of Policy and Regulatory Pathways: Increasing global focus on live biotherapeutic products is driving regulatory agencies (e.g., China's NMPA) to establish clearer approval guidelines, opening pathways for innovative therapies.

Social & Cultural Heritage Narrative: In communities like Mongolian and Tibetan groups, the project can transcend commercial value, elevating to a social issue of "preserving dietary culture," potentially gaining support from government and cultural institutions and enhancing brand image.

Threats

Core Risks of the Technology Platform: Project success critically depends on the precision of gut-specific promoters, the efficacy of adhesin proteins, the efficiency of the lactose metabolic pathway, and the reliable operation of the AI-2/ccdB system in humans. Failure in any single link could lead to a sharp decline in overall efficacy.

Competition from Alternative Technologies: In the long term, in vivo gene editing therapies, should they achieve breakthroughs in safety and cost, could pose a disruptive threat. In the near term, more efficient exogenous lactase preparations or superior traditional probiotics will also compete for market share.

Stringent and Uncertain Regulatory Environment: As a completely novel product category, regulatory agencies may demand data far exceeding常规 standards. Any changes in regulatory policy could significantly delay the product's上市 timeline.

Reimbursement Resistance from Healthcare Systems: Hospitals and medical insurance payers may resist including the product in reimbursement formularies due to its potentially high initial cost, severely limiting market penetration speed. Acceptance and prescribing willingness among physician groups for this novel therapy also require time to cultivate.

Public Opinion and Ethical Controversies: Public debate and negative media reports concerning "genetic modification" and "designing life" could trigger unnecessary regulatory scrutiny and market panic, impacting brand reputation and consumer trust.

Following the internal assessment via SWOT analysis, we further employ the PESTEL model to scan the macro-environmental context surrounding the project. This analysis aims to systematically identify key external factors across the six dimensions—Political, Economic, Social, Technological, Environmental, and Legal—that influence the commercialization pathway of our genetically engineered probiotic therapy, thereby providing a more comprehensive basis for strategic decision-making.

Political

Supportive Industrial Policies: China and many other nations have designated the bioeconomy and synthetic biology as strategic emerging industries, exemplified by China's "14th Five-Year Plan" for Bioeconomic Development. This offers potential support in R&D funding, tax incentives, and science park access.

Stringent Pharmaceutical Regulatory Pathway: As a live biotherapeutic product, it must undergo rigorous approval processes by agencies like the National Medical Products Administration (NMPA). The review standards and speed for innovative therapies by regulators are critical political variables affecting the product's timeline to market.

Influence of the Healthcare Security System: Inclusion in national and provincial reimbursement drug lists depends heavily on government-led health technology assessment and医保 negotiation policies, which will directly determine the product's market access and pricing.

Economic

Health Consumption Upgrade Among the Middle Class: In target markets (especially East Asia), the expanding middle class demonstrates growing willingness and ability to invest in health, creating a payment foundation for high-value, fundamental treatment options.

Supply Chain Costs and Stability: The cost and stability of the supply chain for reagents, equipment required for genetic engineering, and GMP-compliant manufacturing outsourcing services will directly impact production costs and profit margins.

Capital Market Favorability Towards Biotech: Global capital markets maintain strong interest in disruptive biotechnology companies, providing a relatively favorable economic environment for project financing activities.

Social

Public Health Awareness and Precision Nutrition Trends: Consumers are increasingly focused on personalized health solutions and show greater acceptance for innovative therapies that fundamentally improve quality of life.

Perception and Acceptance of Genetic Engineering Technology: Public understanding of the "GMO" concept is polarized, with some segments harboring reservations. Thus, proactive, transparent science communication and public education are crucial for building social trust.

Preservation Needs of Specific Dietary Cultures: In communities like Mongolian and Tibetan groups, the project addresses not only health issues but also aligns with the emotional need to preserve traditional dietary cultures, forming a strong basis for social acceptance.

Technological

Rapid Advancement of Synthetic Biology Technologies: The increasing maturity and decreasing cost of gene editing, synthesis, and genetic circuit design technologies provide powerful drivers for project R&D and future iterations.

Deepening Microbiome Research: A growing understanding of human gut microbiota functions provides a solid scientific foundation and design guidance for assessing engineered bacterial colonization, functionality, and host interactions.

Convergence with Related Technological Fields: Advances in bioinformatics, AI applications in drug design, and novel drug delivery technologies could all contribute to the optimization and success of the project.

Environmental

Environmental Biosafety Regulations: As a genetically modified organism, the research, development, production, and post-use waste disposal of the engineered bacteria must comply with strict biosafety regulations to assess and control potential impacts on the natural ecological environment.

Potential for Sustainable Production: Compared to traditional chemical drug manufacturing, which can generate pollution, microbial fermentation production is more sustainable, aligning with green manufacturing trends and potentially enhancing brand image.

Legal

Intellectual Property Protection Regime: Strong patent protection is the project's core lifeline. A global patent portfolio covering gene sequences, the strain, uses, and manufacturing methods must be established in key markets to build legal barriers.

Biosafety Laws and Ethical Review: The project must strictly adhere to laws like the "Biosafety Law" and pass rigorous scientific ethics reviews, ensuring full compliance from laboratory research to clinical trials.

Advertising Law and Consumer Protection Laws: All promotional claims regarding product efficacy must be truthful and evidence-based during marketing, avoiding violations of medical advertising laws and preventing legal risks associated with misleading consumers.

Lacbutler has also systematically investigated optimal formulations for storage and transportation, aiming to overcome the limitations of traditional biological agents and access broader markets. In resource-limited remote rural areas, employing spray-dried microcapsule technology as an alternative to lyophilized powder can further enhance stability and portability. Particularly in extreme environments such as military operations and disaster relief, medical personnel can quickly mix the microcapsule formulation with carried drinking water for immediate physiological function support.

The preparation of spray-dried microcapsules has been experimentally validated with significant results. Key parameters, including encapsulation efficiency, survival rate, and storage stability, demonstrated excellent performance in accelerated tests. After 6 hours of digestion in simulated gastrointestinal fluids, the decrease in viable bacterial count was controlled within 2.42 log CFU/g. Furthermore, after 28 days of storage at 25°C, the probiotic survival rate remained above 79.87%. These metrics are crucial for maintaining the colonization and therapeutic function of the engineered bacteria.

This project will optimize based on this microencapsulation technology platform and plans to further enhance performance by adjusting wall material ratios and refining process parameters. The interpenetrating network structure constructed using the Salecan-EGCG-Fe³⁺ coordination system not only provides a physical barrier but also, due to its inherent antioxidant properties (DPPH radical scavenging rate of 91.12%), can create a more favorable colonization environment for the engineered bacteria within the intestine. Future work will explore composite uses with other food-grade materials to reduce production costs while maintaining functionality, providing reliable technical support for the commercial application of engineered probiotics.

Our project encompasses several breakthrough technological innovations, primarily including: directed evolution technology for lactase engineering, gut-specific promoter regulation systems, the AI-2 quorum sensing and ccdB toxic protein population regulation circuit, and enhanced gut colonization technology for engineered probiotics.

During the iGEM competition phase, we completed preliminary intellectual property protection through laboratory notebooks and data archiving. To secure the commercial value of the technology, we have formulated the following systematic patent protection plan:

Short-term (Within 6 months post-iGEM):

File 2-3 core invention patents covering: Specific amino acid sequences of the engineered lactase Genetic circuit design of the quorum sensing regulation system Construction method and pharmaceutical use of the engineered bacterial strain

Medium-term (2025-2026):

Based on laboratory data, file patents for the production process and formulation. Pursue international patent applications via pathways like the PCT (Patent Cooperation Treaty).

Long-term (2027 and beyond):

Establish a comprehensive patent portfolio to form a technological barrier. Continuously monitor potential infringement risks. Explore business collaboration opportunities through patent licensing.

Undoubtedly, Lacbutler will progress through multiple phases across various aspects of the project, including system refinement, intellectual property protection, and animal and clinical trials. Each phase involves distinct timelines and testing requirements. Therefore, we have meticulously scheduled activities for the coming years, with the goal of making the therapy available to patients by 2033.

The R&D phases outlined in the Gantt chart are based on an ideal scenario. However, the development of live biotherapeutic products inherently carries risks of technical failure and delays, which are common in biotechnology startups.

Technical Implementation Risks

Engineered Strain Construction and Functional Verification Risk

After months of genetic circuit design and strain modification, the next step requires experimental validation of various engineered bacterial functions. The primary risk is that genetic circuits successful in vitro may fail to operate stably within the complex gut environment. The quorum sensing system might be sensitive to intestinal factors (e.g., pH, microbial metabolites), leading to imprecise regulation of ccdB toxic protein expression and compromised population control.

Mitigation Strategies: Include designing alternative quorum sensing systems (e.g., AHL-based) or incorporating redundant genetic circuits to ensure functional reliability.

Translation Risk from Animal Models to Humans

Engineered bacteria proven safe and effective in animal models may perform differently in human clinical trials. Colonization efficiency could be lower than expected due to differences in gut microbiota composition.

Mitigation Strategies: Utilize humanized gut models for preliminary testing; establish stricter evaluation criteria in preclinical studies; maintain a library of alternative strains expressing different adhesion proteins.

Production Process Scaling Risks

Challenges in Scaling from Lab to GMP Production

Lyophilization and microencapsulation techniques successful at laboratory scale may encounter reduced efficiency and lower viable cell counts when scaled to GMP manufacturing. Optimizing spray-drying parameters requires extensive validation, and encapsulation efficiency might drop from ~84% at lab scale to below 70% at production scale.

Mitigation Strategies: Engage with CDMOs early for process development; establish pilot production lines; allocate sufficient time and budget for process optimization.

Establishing Quality Control Standards

As a first-in-class therapy, the lack of established quality control standards presents a significant challenge. Reliable testing methods must be developed for key metrics like viability, genetic stability, and functional protein expression.

Mitigation Strategies: Engage in early communication with regulatory agencies; reference standards for similar products (e.g., other live biotherapeutics).

Regulatory and Market Acceptance Risks

Regulatory Pathway Uncertainty

As a live bacterial therapy containing engineered genetic circuits, its regulatory classification and approval requirements are uncertain. It might be categorized as a gene therapy product or a novel biologic, with significantly different regulatory demands.

Mitigation Strategies: Request pre-IND meetings with agencies like the NMPA and FDA early; conduct comprehensive preclinical safety studies; submit application materials in stages.

Public Acceptance of Genetically Engineered Bacteria

Despite potential therapeutic benefits, some consumers may have concerns regarding "genetically modified" probiotics.

Mitigation Strategies: Build trust through transparent science communication, collaborate with healthcare professionals for education, and emphasize the advantages of the built-in safety systems. Prepare a base version lacking the genetic circuit elements as an alternative.

The Lacbutler project aims to provide a fundamental therapeutic solution for the global lactose intolerance population through innovative synthetic biology approaches. Building upon the previous risk analysis, establishing a clear exit strategy is not only crucial for maintaining focused R&D objectives but also key for attracting and safeguarding investor interests. We have outlined multiple exit pathways, each aligned with specific stages of the project's development to optimize the risk-reward profile.

Acquisition by a Large Pharmaceutical or Health Food Corporation

This represents one of the most potential exit avenues during the mid-to-late stages of the project. Once our engineered probiotics demonstrate safety and efficacy in clinical trials, they will hold strong appeal for two main types of acquirers:

Large Pharmaceutical Companies

For firms focused on digestive and metabolic diseases (e.g., AbbVie, Takeda) or those with robust OTC product pipelines (e.g., Johnson & Johnson), acquiring Lacbutler would provide a disruptive live biotherapeutic product, enabling them to rapidly capture a significant, long-overlooked multi-billion dollar market for lactose intolerance. It would also demonstrate their commitment to embracing cutting-edge biotechnologies and pioneering new treatment paradigms.

Health Food and Probiotic Giants

For companies deeply invested in gut health and functional foods (e.g., Nestlé Health Science, Danone, Yili), acquiring Lacbutler would signify an upgrade of their product portfolio from "symptom-relieving" foods or supplements to "disease-modifying" therapies, creating a generational advantage and establishing a high technological barrier and brand prestige in the consumer health market.

This strategy typically becomes most attractive after successful completion of Phase II clinical trials, once proof-of-concept is preliminarily established. A prerequisite for success is the strong alignment of our technology platform and future vision with the acquirer's long-term development strategy.

Patent Technology Licensing and Co-development

This constitutes a more flexible, asset-light exit model that can be realized earlier. Once we establish a robust patent moat around the engineered strain's genetic circuits (e.g., the AI-2 quorum sensing system), specific promoters, and formulation technologies, technology licensing becomes a preferred option.

We could license commercialization rights for specific regions (e.g., Greater China or Southeast Asia) to local pharmaceutical companies, or grant rights to develop the technology platform for indications beyond lactose intolerance to specialized biotech firms.

This approach allows our team to focus more on core technology platform iteration and expansion research without bearing the enormous costs and complex management associated with large-scale production and global marketing. By clearly defining licensing fees, sales royalties, and respective rights, this model can generate sustained, stable cash flow for the company within a few years after proof-of-concept is achieved.

Sale to a Biotechnology-focused Investment Fund

This represents another prime pathway for value maximization at the project's mature stage. Globally, numerous private equity or venture capital firms specialize in long-term investments in breakthrough technologies addressing major health challenges.

For them, the Lacbutler project tackles a global public health issue affecting billions, implying immense market potential. A successful, scalable treatment solution promises substantial investment returns.

The key to attracting such investors lies in a detailed commercialization roadmap and a clear scalability analysis, explicitly showcasing the comprehensive plan from clinical trials and regulatory submissions to market launch, along with capital requirements. When the project advances to Phase III trials or the eve of regulatory approval, its value increases significantly. A sale to a biotech fund seeking controlling investments at this stage can deliver substantial returns for all early investors.

To clearly delineate the value cycle and advantages of our Lacbutler product, our team has developed a production timeline for our startup. This outlines the approximate time required to produce 100 units of the drug, considering we are a startup with 7 members/employees. The timeline is approximately 28 days, excluding any post-production validation experiments.

To better analyze our costs and determine our cash conversion cycle, we have established a research and development (R&D) cost breakdown for the on-site production of 10 grams of product under previously defined ideal conditions.

Based on the project development plan, we have formulated an operating expense forecast for the years 2025-2028. The forecast assumes: successful completion of Phase I-III NMPA clinical trials, successful establishment of the patent portfolio, and stepwise capacity expansion—a 10-fold increase in production capacity in 2025, doubling in both 2026 and 2027, and maintaining a stable production scale in 2028.

LacButler is more than a startup project; it is a new wave of health technology initiated by Generation Z. We are a group of biotech enthusiasts and programmers in our twenties, driven by a steadfast mission: to end the troubles of lactose intolerance and reshape humanity's positive relationship with dairy products. We firmly believe that a healthy, free gut is a right everyone should enjoy.

LacButler confronts one of the most common yet most overlooked health challenges of our generation—lactose intolerance. This is not just a health issue; it's about cultural heritage and quality of life. We are proving that young people don't just talk about change—we are using cutting-edge synthetic biology and artificial intelligence to build the solutions ourselves. Our battlefield is in everyone's gut, and our vision is to make every meal an enjoyable experience, free from worry.

Our Tools: Intelligent Self-Assessment & Precision Outreach

To efficiently convey our philosophy of freedom, we have chosen this era's sharpest double-edged sword: artificial intelligence and social media.

We have successfully deployed an intelligent diagnostic program—a "Web Doctor." By analyzing users' classic reactions after consuming dairy products, this program enables preliminary self-checking and risk assessment, providing timely insights before users seek professional medical advice. This is not only our first product touchpoint but also the core channel for collecting real-world data and understanding user pain points.

On the communication front, we will create a vibrant, professional, and highly interactive Instagram presence. This will be more than a bulletin board for project updates; it will be a living room for growing together with our community. We will achieve this through:

Engaging Educational Shorts

Using animations to deconstruct the intelligent working principles of our engineered probiotics.

Scientist Daily Vlogs

Showcasing the young team in the lab, demonstrating how we "culture bacteria and nurture enzymes."

"Gut Freedom" Q&A Boxes

Directly addressing community questions to build trust.

We will initially invite KOLs in the health sector, biotech bloggers, food influencers, and the lactose intolerance community to participate. We deeply believe that this topic, relevant to hundreds of millions, will quickly attract pioneering communities eager to support young people using technology to advance well-being.

Beyond Marketing: Building a Deeply Interactive & Trusted Community Ecosystem

Our strategy extends far beyond online communication. While validating the technology and developing the product, we will continuously deepen our online presence and gradually implement offline activities. We plan to organize "Gut Science Workshops" in universities or high schools upon reaching each significant R&D milestone, bringing the charm of synthetic biology and the concept of health management to the scientists of tomorrow. This not only showcases our progress but also strengthens the emotional bond between LacButler and the next generation of innovators.

Our dream is to create a mobile application integrating health monitoring, microbiota management, and community interaction. Users will not only be able to track their own "Gut Freedom" progress but also visually see how LacButler uses the power of technology to bring change to thousands. Our ultimate goal is to build a global community driven by belief, convinced that technology can create tangible value.

I. Commitment to Stakeholder Responsibility

The Lacbutler project consistently adheres to the "Responsible Innovation" principle of synthetic biology as its core guidance, explicitly committing to the health and well-being of the following stakeholder groups: Lactose Intolerant Individuals: Covering groups from high-prevalence populations (e.g., East Asian consumers) to those with special needs (e.g., individuals seeking to preserve dairy-based cultural traditions), meeting diverse needs through personalized formulation design.

Healthcare Systems

Collaborating with hospitals, clinics, and health management organizations to ensure product efficacy and synergy in clinical use.

Regulatory Agencies

Complying with relevant legislation such as the "Biosafety Law," actively participating in regulatory dialogue, and proactively planning compliance pathways.

Public and Society

Addressing societal concerns regarding genetically engineered products through transparency initiatives and public engagement.

II. Product Safety and Biosafety Assurance

To fulfill our legal and ethical responsibilities for user safety, we have implemented a multi-level risk management system: The AI-2/ccdB quorum sensing control system automatically regulates bacterial population density, preventing over-colonization in the gut. Strict avoidance of antibiotic resistance genes, in accordance with synthetic biology biosafety guidelines.

III. Public Communication and Ethical Considerations

Addressing potential public concerns about genetically engineered products, we commit to:

Precise Communication

Clearly distinguishing Lacbutler from "genetically modified foods," emphasizing its legal status as a live biotherapeutic product and its therapeutic purpose to avoid conceptual confusion.

Ethical Controversy Management

Addressing ethical questions related to "designing life" by demonstrating the reliability of the built-in safety systems through science communication; collaborating with bioethics institutions to conduct public dialogue activities.

Transparency Building

Regularly publishing social responsibility reports and disclosing biosafety data and clinical progress.

IV. Accessibility and Affordability Strategy

To achieve technological inclusivity, we adopt the following measures:

Differentiated Pricing Models

Basic Public Health Version: Collaborate with the government for inclusion in medical insurance catalogs. Consumer Health Version: Reach end-users directly through online channels.

Storage and Transportation Innovation

Utilization of lyophilized microcapsule formulations stable for ≥12 months at 25°C, reducing cold chain dependence. Development of portable packaging suitable for rural, remote areas, and emergency rescue scenarios.

V. Environmental Sustainability and Compliance Management

Environmental Risk Prevention

Establish models to assess environmental dissemination of engineered bacteria, monitoring potential impacts from excretion into the natural environment. Design environment-responsive kill switches that trigger bacterial apoptosis in non-target environments.

Green Manufacturing Commitment

Use of biodegradable materials in the production process. Reduce energy consumption per unit of production capacity through process optimization.

IPO Phase Environmental Responsibility Planning

To avoid cutting environmental investments due to performance pressure post-IPO, integrate ESG indicators into the executive assessment system in advance. Establish an independent Biosafety Committee to oversee environmental compliance performance.

[1] 罗光平, 吴丹, 熊万平, 方小微, 赵娟 & 陈鹏飞. 互穿网络结构双歧杆菌微胶囊的构建与性能研究. 中国测试, 1-10. [2] Shaukat, A., Levitt, M. D., Taylor, B. C., MacDonald, R., Shamliyan, T. A., Kane, R. L., & Wilt, T. J. (2010). Systematic Review: Effective Management Strategies for Lactose Intolerance. Annals of Internal Medicine, 152(12), 797-+. https://doi.org/10.7326/0003-4819-152-12-201006150-00241 [3] Fassio, F., Facioni, M. S., & Guagnini, F. (2018). Lactose Maldigestion, Malabsorption, and Intolerance: A Comprehensive Review with a Focus on Current Management and Future Perspectives. Nutrients, 10(11), 1599. https://doi.org/10.3390/nu10111599