2.1.1 Implementation

The purpose of our survey is to be acknowledged of how much do the public know about sialic acid and its biosynthesis, how much they expect from it, and in what form do they accept sialic acid to appear in. We also intend to know potential consumers’ major concerns and accepted areas, while promoting the use of sialic acid.

2.1.2 Reflection

We received 249 replies globally, mainly from Southeastern China.

The geographic distribution pie chart illustrates that Chinese respondents make up the majority from the southeastern coastline while there are minor international contributions from regions like the United States and Japan. In this market, the survey results also show an extremely rare and conspicuous "double low" characteristic: Public awareness stands at extremely low levels because 86% of survey participants have not actively used these products while scarcity of information shows through 60% of respondents who never heard about "sialic acid." These findings reveal that this market stands at its initial phase and possesses significant potential yet requires immediate promotional efforts.

Picture 3 Respondent Demographics and Baseline Market Awareness

Based on the survey results, it appears that the vast majority of respondents (86.13%) have not used products containing sialic acid, indicating low public awareness or availability. Among those familiar with such products, skincare applications are the most preferred (39.78%), followed by beverages and snacks. The primary audience consists of office workers and younger to middle-aged adults (18–50 years old), suggesting that marketing and product development should focus on these demographics with an emphasis on topical skincare formulations.

Picture 4: Analysis of Consumer Purchase Intent and Commercial Viability

A substantial 82.12% of participants selected the choice that none from their family or friend group use these sialic acid products according to the survey results. 58.03% of potential consumers said they would not choose a sialic acid product next time they made a healthcare purchase, suggesting a gap between interest and actual purchase. The most accepted product price range for these products stands between ¥50 and ¥150/piece where 58.32% of potential consumers find it viable. This shows that to achieve commercial success, a mid-range market positioning strategy is required.

Picture 5 Consumer Purchase Intent and Commercial Viability Analysis

In terms of specific applications, the data suggests that consumers have a very clear demand, with 62.41% and 59.12% of consumers having the strongest desire to see them used in adult supplements and children’s supplements, respectively. However, consumers’ greatest concern in adopting these products is their safety (79.56%) and effectiveness (73.72%), which are by far the most common, much higher than price and brand. This chart vividly illustrates the importance of authoritative verification in converting market demand into actual sales.

Picture 6 Key Consumer Concerns and Product Application Preferences

In terms of market potential, the vast majority of respondents held a neutral to positive view, with over 44% expressing "satisfied" or "very satisfied," and another 45.62% describing it as "average." Negative responses were extremely low, indicating a generally positive market outlook. In terms of functional awareness, consumers have a very clear understanding of the uses of sialic acid, with over half (52.63%) believing its primary function is "immunity enhancement," followed by promoting children's development and anti-aging skin care. This provides clear guidance for the product's market positioning and marketing direction.

Picture 7 Market Potential and Perceived Core Benefits

To understand the acceptance of infant formula containing bio-synthetic sialic acid among mothers and families with infants, we conducted this neighborhood survey, which focused on investigating the acceptance of synthetic sialic acid (bird's nest acid) among people of different social statuses and childcare situations. The results are listed as follows:

Key factors influencing acceptance showed significant divergence:

For some of the segments (mothers of 5-year-old girls, mothers in their 50s with children), "certification from an authoritative organization" ranked first and they could try it if it was proven to be as safe as natural products. Mothers in particular wanted to try the formula on adults first and felt that if products met national standards, they were generally safe. Foreigners and middle-aged women with 12-year-old girls were particularly concerned about the cost and preferred low-priced synthetic products if they had the same effect.

Resistance was also prominent:

Supermarket salespeople with biology background, babysitters, and many parents rejected synthetic products due to the stereotype that "synthetic products equal tech and trickery." Some parents, while vaguely aware of the differences between synthetic and natural ingredients, explicitly expressed their opposition due to lack of understanding or safety concerns. In the specific opinions, an 8-year-old girl would accept milk powder containing sialic acid "because biosynthesis is better than chemical synthesis". Other parents say, "we will not allow our children to take nutritional supplements, but we are fine with adding sialic acid to milk powder."

In general, consumer acceptance of synthetic sialic acid is greatly affected by certification by authorities, price, preconceptions, and form, leading to large differences in perceptions.

Picture 8 The team members are interviewing passers-by

Picture 9 The team members are interviewing passers-by

Picture 10 The team members are interviewing passers-by

Picture 11 The team members are interviewing passers-by

Picture 12 Group photo of team members during offline random passerby interviews

Expert Background

To better explain our product in future outreach and highlight the advantages, challenges, and unique features of biosynthetic sialic acid, we interviewed Professor Yang Dong, he obtained his Ph.D in Biochemistry from University of Maryland-College Park, and completed postdoctoral research in Molecular Biophysics and Biochemistry at Yale University. Currently he holds the position of associate professor at China Agricultural University. His expertise in food biochemistry provided valuable insights into the scientific and technical issues that shape the development of our project.

We primarily wanted to investigate the following technical issues during the interview:

• Confirm technical feasibility and receive expert opinion on major bottlenecks in E. coli‑based sialic acid synthesis (by‑products, endotoxins, and transport limitations in whole‑cell catalysis).
• Recognize upstream strategies to control overflow metabolism (e.g., acetate) and sustain culture health and productivity.
• Clarify practical options for minimizing endotoxin risk (at strain and purification levels) to satisfy safety expectations for nutrition/health applications.
• Assess practical comparisons between whole‑cell catalysis and purified‑enzyme systems (mass transfer, sustainability, and costs), including what optimizations are most important.
• Receive guidance on how to position a biosynthetic product for adoption (efficiency, sustainability, dose benchmarking to human milk, and consumer‑friendly packaging).
• Stress‑test the team’s specific plan (co‑expression of AGE and NAL, fusion/self‑assembly into multi‑enzyme complex, and tuning of enzyme ratios) against real production constraints.

Key Points Discussed in the Interview

The main obstacles in the further optimization of E. coli for sialic acid synthesis are by-product inhibition and endotoxins. In particular, the overproduction of acetate under carbon excess conditions decreases the pH of the culture and subsequently the growth of the cells. Moreover, other by-products, including lactate and ethanol, also disturb the metabolic homeostasis. Endotoxins, essential components of the outer membrane of E. coli, have a high level of immunogenicity and are harmful to product safety and enzyme activity.

Multiple strategies can be applied to overcome these limitations, such as the knockout of the genes that leads to the synthesis of acetate, and overexpression of genes that allow for the increased reutilization of acetate. The design of engineered strains that can degrade the toxic endotoxins also effectively improves the product quality. The endotoxins can also be removed using membrane filtration and adsorption gels during the downstream purification, further improving the safety and yield.

Whole-cell catalysis is another strategy for the production of sialic acid that also has great potential. It is more sustainable and cost-effective but there are also its bottleneck including the limited transport of substrates, which can be further addressed by specific optimizations. The commercialization of biosynthetic sialic acid can focus on its efficiency and sustainability as the key selling points. Dosage can also be determined based on the established standards in human milk, and the packaging with high recognition can also be applied to ensure consumer acceptability.

Conclusion and Reflection

• Chassis and endotoxin plan

We are using E. coli BL21(DE3), so we will treat endotoxin control as a core design choice. We will include a routine endotoxin check (chromogenic LAL kit) in our workflow and set an internal safety target appropriate for oral nutrition use. We will prototype a simple downstream removal step (membrane filtration + anion-exchange resin), and we will evaluate a low-endotoxin strain option in parallel if time allows.

• Whole-cell catalysis aligned with transport

Our multi-enzyme design aims to channel ManNAc from AGE directly into NAL. To match this, we will decide on where catalysis happens: cytosol (baseline), periplasm (via signal peptides), or gently permeabilized “resting cells.” We will compare these formats for substrate/product transport by measuring initial rates with GlcNAc + pyruvate feeds and checking whether native metabolism diverts intermediates.

• Protecting substrates and product from native pathways

We will map host routes for GlcNAc/ManNAc/Neu5Ac (e.g., uptake and catabolism) and reduce competition by design. Our first-line approach is compartment choice and cultivation conditions; if needed, we will test CRISPRi to knock down key competing steps (e.g., Neu5Ac uptake/catabolism) rather than full knockouts, to keep the build lightweight.

• Making our multi-enzyme complex testable and tunable

We will compare two architectures: (1) direct fusions with flexible linkers and (2) modular self-assembly (e.g., SpyTag/SpyCatcher or scaffold domains). We will predefine a small matrix of AGE:NAL stoichiometries (1:1, 1:2, 2:1) using promoter/RBS strength and/or scaffold valency.

To verify in-cell ratios, we will attach fluorescent tags (e.g., sfGFP on AGE, mCherry on NAL) in diagnostic constructs and correlate fluorescence ratios with activity.

• Managing overflow metabolism early

To reduce acetate, lactate, and ethanol, we will (i) prefer glycerol or controlled glucose feeding over rich glucose, (ii) maintain good aeration, and (iii) monitor pH and acetate using simple enzymatic kits. We will document how these policies impact growth, enzyme expression, and Neu5Ac productivity.

• Reusability and sustainability

We will evaluate whole-cell catalyst reuse across multiple batches, measuring retained activity each cycle. If activity holds, we will include this as a sustainability and cost advantage in our outreach.

• Communication and adoption

We will connect our technical choices to user-facing benefits: consistent quality, safety management from the start, and a lower environmental footprint than chemical synthesis. For dosage communication, we will reference the range present in human milk and explain how our biosynthetic route enables controlled, reproducible levels.

Picture 13 Interview with Prof. Yang

Picture 14 Interview with Prof. Yang

Expert Background

We interviewed Ms. Cui Binhui to gain insights from an industry analyst on the market potential, consumer trends, and commercialization pathways of sialic acid in the food and related sectors. Ms. Cui holds a Master’s degree in Commerce from the University of Sydney and is an ACCA member. She currently serves as a senior industry analyst at a publicly listed domestic company, where she specializes in evaluating industry trends, conducting in-depth analysis, and valuing covered companies and equities. Her areas of expertise include agriculture and the leisure food industry.

We primarily wanted to investigate the following issues during the interview:

• To understand the market structure and growth trajectory of sialic acid (global concentration vs. rapid domestic growth) to validate opportunity and timing
• Clarify commercialization pathways: B2B ingredient vs. consumer formats, pricing dynamics, capacity ramp and supply security
• Map regulatory routes for food use and export (China, US, EU), and how standards influence product specs, QA and labeling
• Identify differentiation beyond cost—purity, safety, sustainability, traceability—and how to raise consumer awareness
• Connect process choices (whole-cell catalysis, enzyme assembly) to COGS, scalability and competitive positioning
• Learn how to leverage exhibitions, partnerships and data-driven (AI) tools to accelerate adoption

Key Points Discussed in the Interview

The international sialic acid market shows oligopolistic characteristics through limited market dominance by several enterprises while the domestic market remains in rapid expansion with most companies only developing sialic acid at this stage.

With regard to profitability, the gross margin of sialic acid is relatively high. In terms of technology, large-scale production technology abroad has matured; domestic companies are still at a bottleneck despite breakthroughs in ton-level production; artificial synthesis is currently mainly fermentation of E. coli with specific carbon sources; enzymatic synthesis and other optimization methods are being explored

From a supply and pricing perspective, security depends on raw material availability and production capacity. Due to regulatory constraints, chemical synthesis methods may hold advantages. Domestic companies have already achieved small-scale production, with prospects for capacity expansion in the future. As costs decline, prices are expected to fall, potentially sparking price wars and reshaping industry dynamics.

On industry prospects and market promotion, growth opportunities lie in nutrition upgrades, the maternal and infant sector, and the aging population, though consumer awareness must be improved. The product is already legally approved as a food additive, though regulatory systems still need refinement, and future standards are expected to align with those in Europe and the U.S. Export competitiveness will depend on technological innovation and cost-effectiveness, while re-export trade may help bypass certain tariffs. Although artificial synthesis has cost advantages, low consumer recognition remains a barrier to market adoption.

For commercialization and production planning, sialic acid is expected to be developed into powder form for reprocessing, with business strategies supporting manufacturers in expanding market share. Whole-cell catalytic methods show promise but face bottlenecks; exhibitions, AI, and other tools may help optimize production and accelerate promotion. Overall, the industry presents both opportunities and challenges in terms of markets, technology, supply, and demand, requiring joint efforts to advance sustainable development.

Conclusion and Reflections

Where we will play first

We will position our product as a B2B, food-grade sialic acid ingredient for nutrition and functional food applications, with a stretch goal of meeting stricter specs suitable for infant and elderly nutrition. This lets us focus on quality, consistency, and documentation while building partnerships with formulators.

Product form and target specifications

We will develop a free-flowing powder suitable for blending. Our initial internal spec sheet will include: assay/purity (HPLC), identity (UV/FTIR as available), residual substrates (GlcNAc, pyruvate), inorganic ions/metals, moisture, microbiological limits, antibiotic residues (not detected), host DNA/protein (below internal threshold), and endotoxin (internal safety target for oral use). We will add simple, validated methods to our lab workflow (resorcinol assay/HPLC for Neu5Ac; chromogenic LAL for endotoxin; plate counts for bioburden).

Packaging concept: moisture/oxygen-protective pouches or HDPE/foil-lined drums; we will run mini stability studies (40°C/75% RH and room temperature) to define shelf-life claims.

Linking market insight to our engineering plan

To compete as prices decline, we will focus on yield, productivity, and simplified downstream. Our AGE–NAL co-localization strategy is directly tied to cost: higher conversion of GlcNAc and reduced intermediate losses lower COGS and waste.

We will track three metrics per run: titer (g/L), overall yield (mol Neu5Ac per mol GlcNAc), and volumetric productivity (g/L/h). Our goal during iGEM is to demonstrate a clear improvement over non-colocalized controls, and to reduce downstream steps required to reach spec.

Regulatory readiness and export mindset

We will map regulatory pathways early:

China: align with current GB standards and the “new food raw material”/additive frameworks; ensure compliant labeling and documentation.

US/EU: outline GRAS/NDI (US) and Novel Food (EU) possibilities depending on intended use, and design our process to minimize host-derived residuals to support dossier preparation.

We will not make disease-related claims. Our communication will focus on quality, consistency, and alignment with naturally occurring levels (e.g., the range present in human milk), subject to local regulations.

Differentiation beyond price

Quality: emphasize high purity, low residuals, and robust batch-to-batch consistency. We will publish a spec sheet and representative batch COA-style data on our wiki.

Safety-by-design: integrate endotoxin control at the chassis/process level and add a polishing step; document our testing protocol.

Sustainability and traceability: describe our fermentation-based route, planned catalyst reuse tests, and a simple material/energy footprint snapshot. We will pilot QR-enabled batch traceability on the wiki.

Supply security and inputs

We will review sourcing options for GlcNAc and pyruvate, compare cost/availability, and note alternatives (e.g., upstream conversion routes) as future work. We will articulate how our process tolerates feedstock variability (quality specs for inputs).

Go-to-market building blocks

Pilot milestones: lab proof-of-concept meeting our internal spec; small pilot with process reproducibility across at least three batches; a basic cost model identifying the main levers (substrate cost, conversion yield, cell reuse, downstream resin/membrane load).

Partner plan: engage with formulators and toll manufacturers at key events (e.g., FIC China, Hi & Fi Asia-China, Vitafoods/Hi Europe, SupplySide) to gather requirements and iterate specs.

Documentation pack for partners: tech data sheet, sample COA, safety and stability summaries, and a process overview noting GMO control and removal of processing aids.

Communication and consumer awareness

We will use clear, non-technical visuals to explain what sialic acid is, how fermentation produces it, and how our process ensures quality and safety. We will avoid overpromising efficacy and focus on measurable product attributes (purity, consistency) and responsible dosing grounded in reference ranges from human milk literature.

Risk and resilience

Price competition: we will differentiate on quality, documentation, and service (spec flexibility, technical support) instead of racing purely to the bottom on price.

Scale-up risk: we will design experiments to be directly transferrable (e.g., oxygen transfer considerations, feeding policies) and document scale-sensitive parameters to de-risk pilot runs.

What this changes in our design

We translate market and regulatory insights into concrete specs, tests, and documentation that our lab work will target from day one.

We tie our AGE–NAL assembly and whole-cell strategy to cost and quality metrics that matter for commercialization, not just titers.

We adopt a B2B-first path with a clear partner package, while keeping export and regulatory compatibility in view to future-proof the project.

Picture 15 Interview with Ms. Cui

Background:
To better understand the current industry landscape and future development of industrial production, as well as to analyze the advantages, disadvantages, unique strengths, potential issues, and production challenges of various drug formulations, we interviewed Mr. Ye Mao to gain insights into pharmaceutical formulation R&D and production. Mr. Ye obtained his master's degree in Pharmaceutical Engineering from Wuhan University of Technology in 2008 and has extensive experience in the pharmaceutical industry. Currently serving as the production head at ApicHope Pharmaceutical Group Co.,Ltd, an innovative listed pharmaceutical company in Guangzhou, he has rich expertise in the R&D and production of multiple drug dosage forms, which helps us identify key challenges in project implementation.

We primarily wanted to investigate the following issues during the interview:

• Understand how industrial producers choose and scale dosage forms, and how those choices affect quality, cost, and compliance.
• Compare fermentation vs. new enzymatic/one‑pot approaches in terms of impurity risk, purification burden, and safety.
• Clarify specification and regulatory expectations across grades (industrial vs. food vs. pharma), including impurity controls (e.g., heavy metals, unknown impurities, residual proteins/endotoxin).
• Learn practical pathways for commercialization in Asia vs. US/EU, where awareness and acceptance differ.
• Translate pharmaceutical Quality by Design (QbD) thinking into our food‑grade biomanufacturing plan.
• Explore university–industry collaboration models to accelerate pilot production and market entry.

Key Interview Points:
As an endogenous substance in humans, sialic acid is primarily synthesized in the liver. Its common forms include:

• N-acetylneuraminic acid (key for brain development)
• N-glycolylneuraminic acid (requires external intake from sources like bird's nest, eggs, and milk, with content ranging from 7% to 12%)

As a food additive, sialic acid is highly safe, with China's national standard setting a maximum daily intake of 500 mg. Excessive intake poses no significant toxicity risks, but attention must be paid to by-products and impurities during synthesis.

Traditional fermentation methods for sialic acid production have low efficiency and may introduce impurities like exogenous proteins, potentially causing immune or allergic reactions. New enzymatic or "one-pot" methods offer higher catalytic efficiency, but microbial fermentation may introduce safety risks, requiring strict purification to control impurities (e.g., heavy metals <10 ppm, unknown impurities <0.1%).

Food-grade sialic acid (purity ≥98%) costs about ten times more than industrial-grade and must strictly comply with National Medical Products Administration regulations to avoid safety hazards from industrial-grade materials.

Market-wise, Asia is the primary market due to cultural familiarity with "bird's nest acid," where sialic acid is more readily accepted for benefits like cognitive enhancement and infection risk reduction. The Asian market currently focuses on this region due to historical awareness of bird's nest, while Europe and the U.S. are still in early stages of development due to lack of widespread value recognition.

Over the next five to ten years, enzymatic and genetic engineering technologies will drive sialic acid production toward precision and modularity. Mr. Ye suggested that startups could accelerate commercialization through university-industry collaboration, leveraging academic technological strengths and corporate commercialization capabilities.

Conclusion and Reflection

Our positioning and grade

We will target a food-grade, B2B ingredient first (not industrial grade), aligning with China’s food additive framework and keeping an export mindset. We will explicitly separate our work from pharmaceutical claims and design our documentation for food use.

Draft quality targets (CQAs) informed by the interview

Identity and assay: confirm Neu5Ac identity and content; monitor Neu5Ac/Neu5Gc profile to ensure we supply Neu5Ac only.

Purity: aim for food‑grade purity ≥98% (aligning with current market practice); set “unknown impurities” as ≤0.1% (interview guidance) and monitor known process-related impurities (e.g., residual substrates).

Safety: establish internal limits for heavy metals (e.g., <10 ppm per interview guidance), residual host proteins/DNA (minimized), endotoxin (internal oral-use target), bioburden (within food‑grade limits).

We will summarize these as a draft spec and build a simple certificate‑of‑analysis (COA) template for sample batches.

Impurity control strategy: prevent → minimize → remove

Prevent at the source: use defined media and vetted inputs to reduce metal and impurity introduction; keep animal-derived materials out. Choose a low-endotoxin chassis option if feasible within our timeline.

Minimize by design: our AGE–NAL co-localization aims to reduce intermediate accumulation and side reactions, lowering impurity formation versus non-colocalized expression.

Remove efficiently: outline a high-level downstream path suitable for a small, polar acid (cell removal → protein clarification → selective capture → polishing for endotoxin/trace impurities) without adding complexity that would raise costs.

Safety and dose stewardship

We will reference the Chinese national standard daily intake (up to 500 mg/day) in our communication and avoid any disease-related claims. Our messaging will focus on quality and responsible use consistent with food regulations.

Translating formulation lessons to food formats

Initial product form: a free-flowing powder for blending into foods or beverages. We will prioritize low moisture uptake, good flow, and taste acceptability (plan basic taste‑masking via compatible carriers/flavors).

Packaging: moisture/oxygen protection (e.g., foil-laminate pouches or lined drums) with desiccant; run small stability checks at room and elevated humidity/temperature to guide shelf‑life statements.

Consumer‑facing prototypes (lab demo only): simple sachets or stick-packs and a model beverage spiking study to observe clarity, taste, and short-term stability.

Analytics we will incorporate early

Neu5Ac quantification/identity: colorimetric screening plus chromatographic confirmation when available.

Impurities: track residual substrates/organics by chromatography if accessible; run total protein and nucleic acid screens as simple proxies for host contaminants.

Endotoxin and bioburden: include routine checks suitable for food-grade decision-making.

Process design with QbD mindset

Define our critical quality attributes (CQAs) and map them to critical process parameters (CPPs) such as feed strategy and oxygenation (documented qualitatively for transferability).

Record scale‑sensitive factors (mixing/aeration) during lab runs so we can hand partners a clearer pilot brief.

Include controls that demonstrate the value of our AGE–NAL assembly (same-cell/no‑assembly vs. assembled complex) and tie improvements directly to yield, productivity, and impurity reduction.

Market focus and education

Short term: concentrate on Asia where awareness of “bird’s nest acid” is higher, and frame our product as “fermentation‑derived Neu5Ac with tight impurity control.”

Mid term: adapt communication for US/EU with simple explanations (what Neu5Ac is, where it naturally occurs) and emphasize quality, safety, and traceability rather than efficacy claims.

Collaboration pathway

We will pursue a university–industry collaboration to (i) validate our draft spec with an experienced manufacturer, (ii) get feedback on scalable downstream options, and (iii) explore toll‑manufacturing for small pilot batches when data support it.

Risk guardrails we adopt now

Industrial‑grade adulteration risk: we will set minimum acceptable specs and supplier qualification rules for all inputs; we will not position or source “industrial grade” for food use.

Price pressure: our technical focus (higher conversion, simpler polishing, potential whole‑cell reuse) is chosen to defend cost while maintaining quality.

Acceptance gap outside Asia: we will avoid overpromising benefits and instead highlight measurable attributes (purity, consistency, safety testing).

What this changes in our design

We formalize a food‑grade spec and COA mindset from the start, tying our engineering choices to impurity control and safety testing.

We link our enzyme assembly benefits not only to titer but to reduced impurity burden and simpler downstream, which matter for commercialization.

We adopt practical formulation and packaging directions suitable for a B2B ingredient today, while keeping options open for future consumer formats with partners.

Picture 16 Interview with Mr. Ye

Expert Background

To address key challenges in our sialic acid (SA) synthesis project—including multi-enzyme system host selection, reversible enzyme reaction optimization, industrial scale-up, and market positioning—we interviewed Prof. Oliver Yu. Prof. Yu is Co-Founder/Chief Strategic Officer (CSO) and a Affiliate Faculty at MIT. He has profound expertise in food science, synthetic biology, and "lab-to-table" biomanufacturing (backed by Aquafarmtory’s experience), which aligns with our project’s focus on efficient SA biosynthesis via E. coli BL21(DE3)-based multi-enzyme complexes and its commercial translation. His insights help solve our technical and practical dilemmas.

Interview Takeaways

(1) Technical R&D of SA Synthesis

Prof. Yu focused on host selection, reversible reaction optimization, and multi-enzyme efficiency evaluation. For E. coli vs. Pichia pastoris, he noted E. coli BL21(DE3) has fast growth, low metabolic burden, and mature fermentation protocols (suitable for SA synthesis as AGE/NAL enzymes need no complex modifications), while Pichia excels in protein folding but has higher costs. He recommended prioritizing E. coli. To drive reversible enzyme reactions, he proposed coupling energy-consuming reactions (e.g., co-expressing pyruvate kinase to consume by-products) or regulating cofactor regeneration, which can integrate with multi-enzyme complexes. Beyond product rate, he emphasized monitoring enzyme stability, substrate specificity, and by-product ratio to guide pathway optimization.

(2) Industrialization of SA Production

Regarding industrial potential, Prof. Yu stated the E. coli-based multi-enzyme scheme is more viable (lower scaffold preparation cost vs. fusion enzymes). Key scale-up concerns include strain stability (using antibiotic-free plasmids) and uniform mixing in large bioreactors. He shared Aquafarmtory’s experience: iterative small-batch trials (100L→1000L) and real-time monitoring (e.g., HPLC for SA detection) to bridge the lab-to-industry gap. For sustainable biomanufacturing, he suggested reusing fermentation medium and optimizing temperature to reduce waste and energy consumption.

(3) SA Market & Business Strategy

Prof. Yu advised enhancing biosynthetic SA’s sustainability advantage by quantifying its carbon footprint (e.g., lower CO₂ emissions than natural extraction) and obtaining third-party certifications. For consumer education, he recommended referencing Aquafarmtory’s alternative protein experience—publishing clinical data and using transparent storytelling to convey "nature-equivalent" safety. He also proposed exploring innovative applications (pet nutrition, plant protection) with Aquafarmtory’s product matrix and suggested joint venture’s with downstream enterprises (vs. sole B2B sales) to unlock market value.

Conclusion and Reflection

After the interview, we clarified our project direction: technically, we will first optimize the E. coli-based multi-enzyme system (e.g., testing AGE/NAL ratio 1:1.2) and integrate energy-coupled reactions. Industrially, we will adopt iterative small-batch trials and real-time monitoring to ensure scale-up stability. Market-wise, we will prioritize sustainability communication and phased application expansion.

This interview reminded us that synthetic biology projects need to balance technical innovation with industrial feasibility and market demand. We will further collaborate with chemical engineers to optimize bioreactor design and consult environmental experts to quantify our carbon footprint, ensuring the project advances practically.

Picture 17 Portrait of Prof. Yu

Picture 18 Professor Yu's offline interview

Picture 19 Professor Yu introduces Aquafarmtory Company

Picture 20 Team photo of visiting Aquafarmtory Company

In order to become more familiar with similar products, technologies, and equipment already available on the market, we engaged in in-depth discussions with exhibitors from both commercial and biological perspectives to understand the industry's views on our project, obtain professional feedback and opinions, and explore potential applications and areas for optimization.

Picture 21 Exhibition interviews

Picture 22 Exhibition interviews

Picture 23 Exhibition interviews

We learned from the booth representative at Tianmu Biotechnology Co., Ltd. that future research will combine AI with biology to empower biotechnology, such as screening for main products to reduce the accumulation of by-products. Biotechnology is currently a trend, strongly supported by the state to move toward high-tech and precision. The field of biotechnology can now provide advanced technical equipment to improve efficiency and experimental accuracy. In terms of commercialization, target users may prioritize safety concerns and be reluctant to accept synthetic sialic acid as a substitute for natural sialic acid. However, in food applications, synthetic production eliminates the need for extraction and further processing, though it may increase the risk of impurity residues, it significantly reduces costs, making it feasible to expand access to lower-income populations.

Picture 24 Group photo of the exhibition reporting team 1

Picture 24 Group photo of the exhibition reporting team 2