Prof. Jee Loon FOO
In light of our pursuit to understand some of the literature we were basing our project upon,
we
reached out to Research Assistant Professor Jee Joon Foo at the National University of
Singapore. We first gave a quick presentation regarding our project scope and explained the
specific parts of our chassis. As an expert in the field of synthetic biology, Professor Foo
gave us feedback regarding our chassis selection as we were still considering the sort of
yeast
model we would use in our project. Additionally, he explained that the processes we were
intending to take were theoretically doable although questions regarding the feasibility
within
the given time were raised.
From our meeting with Professor Foo, we were able to reaffirm the scope, content and
intentions
of our project. Additionally, through the recommendations he provided regarding the choice
of
the chassis (taking into consideration factors such as literature published related to the
chassis, survivability etc.) we were able to narrow our choice down to Y.
lipolytica.
Prof. Yong LAI
Based on the discussion with Professor Yong Lai who is a professor in the School of
Engineering
at the Hong Kong University of Science and Technology (HKUST), we have learned several key
insights regarding the integration and expression of the PUFA synthase cluster in Y.
lipolytica:
The main gene integration method in Y. lipolytica is non-homologous end joining rather
than
homologous recombination, which is less efficient. Professor Lai suggested exploring two
rounds
of transformation to improve integration efficiency. He also highlighted the importance of
checking existing publications to understand the novelty of the project, noting that the
higher
specificity of the PUFA synthase could be a key advantage.
Regarding the integration strategy, Professor Lai suggested exploring random integration
using a
linearized DNA fragment. He also discussed the challenges of integrating the large PUFA
synthase
subunits, which may be difficult due to their length. To address this, he recommended
amplifying
the sequences from the algae through PCR and potentially exploring codon optimization or
mutations.
Professor Lai also provided several alternative strategies, such as using a yeast artificial
chromosome for the recombination and exploring Gibson assembly.
For the expression analysis, Professor Lai highlighted the potential challenges of using
Western
blot for large proteins and suggested exploring alternative methods such as LCMS or
real-time
PCR. He also discussed strategies to improve expression, such as knocking out the PEX10 gene
to
prevent protein degradation.
Overall, the discussion with Professor Lai provided valuable insights and practical
suggestions
to address the key challenges in the integration and expression of the PUFA synthase cluster
in
Y. lipolytica. We could now consider these recommendations to refine their
experimental approach
and improve the likelihood of success in their project.
Prof. David BANFIELD
We also reached out to Professor Banfield, a cell biologist in the Division of Life Science
at
the Hong Kong University of Science and Technology (HKUST). In this outreach, we had the
opportunity to discuss several key considerations regarding enzyme expression and complex
purification. The primary objective was to ensure we have a reliable way to measure the
expression of the target enzymes, as the commonly used His-tag may not be compatible with
the
available antibodies. Alternative options, such as the HA tag and Myc epitope, were explored
as
potential solutions.
To start the investigation, we proposed expressing each gene individually in yeast to better
understand the system. This approach would allow us to consider strategies for purifying the
complex itself, potentially using affinity chromatography with the combination of tag 1 and
tag
2 or peptide sequences.
One potential challenge Professor Banfield pointed out was the concern about the
transformation
process, particularly if the same marker is used for multiple integrated genes. To address
this,
the use of auxotrophic markers and the recycling of markers through the incorporation of
loxP
sites were suggested as possible solutions.
Potential issues, such as cryptic splice sites and the impact of suboptimal codons near the
5'
end of the gene, were also discussed. These factors could potentially lead to premature
termination or detachment of the RNA polymerase, resulting in problems during expression.
Lastly, the challenges associated with yeast transformation, particularly with large
plasmids,
were addressed. The recommendation was to consider separating the genes into smaller units,
as
yeast may have difficulty maintaining and expressing large plasmids. The order of
integration
and potential toxicity issues were also discussed, with the suggestion of trying different
permutations to identify the optimal approach.
This outreach has provided valuable insights and guidance on the various considerations and
potential challenges involved in enzyme expression and complex purification, which will be
instrumental in refining our experimental approach and ultimately advancing our research
goals
which is greatly helpful insight to us.
Dr. Du Fei
We are honored to have received a recommendation from Professor Sun Xiaoman of Nanjing Normal University,
which has referred us to reach out to Dr. Du Fei.
We arranged an interview with Dr. Du Fei from Nanjing Normal University, an advisor from iGEM NNU Team
2022 and published a paper related to Y. lipolytica. During the interview, we inquired about the
impact
of codon bias on the feasibility of cloning directly from Schizochytrium sp., expressing in our chassis,
the constructs in it and how to obtain this strain. Her insights helped us shape our second engineering
cycle.
Prof Joseph WONG
We were able to reach out to Professor Joseph Wong, a synthetic biology professor from the Hong Kong
University of Science and Technology (HKUST), One of the primary concerns discussed was the
strain selection and cultivation process. It was noted that the strain development can be a
time-consuming endeavor, often taking months to achieve the desired characteristics. The
importance of proper biological safety protocols was emphasized.
The discussion also delved into the potential for protein secretion from the cells. While the
codon bias was not seen as a significant concern, we focused more on ensuring proper organelle.
Regarding the Y. lipolytica plasmid, we discussed the potential differences compared to
other
systems and the importance of understanding the inductive or repressive mechanisms involved.
The concept of gene knockouts was also discussed, with the suggestion of exploring the use of
repressive promoters and evaluating the conditions, such as redox and pH, that could influence
the desired outcomes. We also highlighted the potential use of resources like the Saccharomyces
Genome Database (SGD) to gather information on genetic interactions, physical interactions, and
synthetic lethality.
Throughout the discussion, we considered various aspects of the project, including the production
and secretion of the desired compounds, the growth characteristics and culturing densities of
Yarrowia lipolytica, and the potential pathways involved in the synthesis of long-chain
hydrocarbons.
By engaging with Professor Joseph Wong and leveraging his expertise, we were able to gain a deeper
understanding of the challenges and opportunities inherent in the project. This collaboration
has helped to refine the project's direction and inform the necessary adjustments to ensure the
successful development and implementation of the desired microbial engineering solutions.
Prof. Rachubinski
From left to right. First row: David, Andrew, Kathy; Second row: Selina, Prof.
Rachubinski, Aurora; Third row: Angel
We are honoured to be able to reach out to Professor Rachubinski, who is the professor
from the University of Alberta. He enhanced our understanding the Role of PEX10 in Lipid
Accumulation in Y. lipolytica.
PEX10 is an essential protein in the biogenesis of peroxisomes in the yeast Y. lipolytica.
It
plays a crucial role in the import of matrix proteins, such as PEX2, PEX10, and
PEX12, into
the peroxisome. In contrast, genes like PEX3 have an even more fundamental function, as
the absence of PEX3 results in the complete lack of peroxisomes.
To increase the percentage of DHA production rather than accumulating other unwanted
fatty acids, our strategies focus on guiding the fatty acid synthesis pathway towards DHA
production, limiting the enzymatic capacity for short-chain fatty acid synthesis.
Modeling the effects of PEX10 knockout on fatty acid synthesis in Y. lipolytica is challenging,
as PEX10 is not directly involved in metabolic reactions. Consulting experts in metabolic
modeling, such as Professor Paul LaPoint and Dr. Pred Mast, may be beneficial in
developing appropriate modeling strategies.
To validate the PEX10 knockout, key steps include isolating genomic DNA, performing PCR
to check the 5' and 3' ends of the gene, comparing the knockout strain to the wild-type
control, and using immunofluorescence with a PTS1 antibody to visualize peroxisome matrix
protein import.
This outreach to Professor Rachubinski provided valuable insights into the role of PEX10 in
Y. lipolytica and the considerations for optimizing DHA production through genetic
engineering. The information gathered will guide our team's ongoing research and
experimental design.
Dr. Young-Kyoung Park
We had an interview with Dr. Young-Kyoung Park from Micalis Institute, France, an experienced researcher
in Yarrowia lipolytica bio-production. As an insider, she provided us valuable insights into the
current
industry of Yarrowia lipolytica bioproduction. She pointed out that one of the main obstacles for
current Yarrowia lipolytica production is still the lack of fundamental knowledge and genetic
tools for
this organism and its application, for example, the inducible promoters. Though a wide range of promoter
studies has been performed, the ones focused on inducible promoters is still quite few. She agreed that
our strategy for stage controls and improving promoter design would be effective, and the
characterization of inducible promoters would be valuable.
We also discussed the regulation regarding GMO. She is not aware of any existing products produced by
genetically modified Yarrowia lipolytica directly serving as food use, and a controversial and
long
approval process is involved. However, from her opinion, it would be fine to use the genetically
modified organism for food or supplement production, as no known harm can yet be identified.
Lastly, Dr. Park also gave us some useful suggestions about Yarrowia lipolytica transformation as we were
using the protocol she documented in one of her papers. We confirmed many details regarding the
experimental steps including cultivation conditions etc. and she helped us with troubleshooting. She
also shared her experiences on transforming Yarrowia lipolytica, indicating that our transformation
efficiency still has much space for future improvements, which we are currently working on.
Prof. Rodrigo Ledesma-Amaro
Through email, we are glad to have interviewed Prof. Rodrigo Ledesma-Amaro from Imperial College London,
one of the top Yarrowia lipolytica research experts in the world.
Regarding the chassis choice, Prof. Ledesma-Amaro provided us with successful Yarrowia lipolytica
industrial applications to look into such as Ginkgo, and pointed out that the chassis’s ability to grow
from low cost substrates and waste represents one of its biggest advantages, adding towards the
motivation of our chassis use. We also learned more about the efflux process as proposed in one of his
papers, and confirmed that it will greatly facilitate the purification process as sometimes lipids
floating can be seen, ready for collection. The feedback gave us a better understanding of the efflux
strategy and contributed to its establishment in our design.
We also consulted him about the transformation of Yarrowia lipolytica, and received valuable
feedback on
setting the control groups and troubleshooting. With his advice, we are finally able to demonstrate a
successful transformation.
Prof. Chen Zhi
We are honored to talk to Prof. Chen Zhi from China Agriculture University expertise in metabolic
engineering of DHA-producing microalgae Schizochytrium sp.. In the meeting, a detailed discussion
was
carried out regarding the current state of affairs and metabolic engineering in Schizochytrium
sp.,
which shares similarities with our own projects. The conversation first delved into the significance and
innovativeness of introducing PUFA synthase pathway into Yarrowia lipolytica.
Challenges were raised concerning DHA production in microalgae versus yeast, particularly in balancing
the levels of unsaturated fatty acids like DHA and enhancing triglyceride storage to increase their
proportion, which is meeting a bottleneck in Schizochytrium sp. According to Prof. Chen. The need
for a
balance between saturated and unsaturated fatty acids, e.g., for membrane integrity, was emphasized
alongside boosting production yields. She also highlighted the potential of optimizing PUFA synthase
according to domain functions for enhanced product specificity in the future.
The risk factors discussed revolved around experimental outcomes, emphasizing the need for sufficient
enzyme levels for successful production in oil-producing microorganisms. The conversation further
touched upon metabolic engineering strategies involving gene knockout techniques for enhancing
production and the selection of pathways for engineering, where we realized that microalgae engineering
is actually taking much reference to Yarrowia lipolytica studies, as there is still a wide gap
between
research powers devoted to the 2 species worldwide. The maturity of genetic editing tools in microalgae
and yeast was compared, with Yarrowia lipolytica being deemed more mature with more applications,
serving as a solid evidence of why our project is beneficial to the current industry.
We further consulted Prof. Chen about the cultivation of microalgae and yeast, with a focus on salt
requirements differing between the two types of microorganisms, alongside measures to prevent
contamination during the fermentation process. We also explored the potential for industrial scale-up
and the considerations surrounding regulatory compliance and safety standards in the production of
PUFA-rich oils for nutraceutical and food applications.
Prof. David Silver
In hope to elucidate the DHA transport mechanism for the efflux strategy in our project, we reached out
to Prof. David L. Silver from Duke-NUS Medical School, expert in PUFA-specific membrane transporters.
During the meeting, we discussed the potential of increasing DHA yield through efflux strategies. From
the conversation we learned that in the existing case of fatty acid efflux as free fatty acid,
non-transport mediated diffusion due to high membrane permeability to fatty acids is likely the main
driving force. This method has its advantages, but due to the high instability of DHA and resulting cost
of the downstream process, exporting free DHA is not meeting the needs of the industry. Instead,
triacylglyceride or phospholipid forms for cost-effective and stable DHA products were suggested.
After hearing about our project, he pointed out that photosynthetic microalgae cultivation in large
outdoor ponds is space-intensive and costly, in contrast our method may operate in cities at various
scales with less resources or interference with the environment, indicating that it is potentially more
sustainable and cost-effective.
Further discussions centered on the transporter efficiency. We delved into high capacity transporters
like LPC-transporter Mfsd2a, which can reach a very high transport flux but only in charge of the PUFA
influx and is unlikely to be inverted. He hypothesized that the specificity of the transporter is
probably due to the higher spatial fluidity of PUFA over other fatty acids. We also mentioned the
potential of using ABC transporters for efflux.
Additionally, the fate of absorbed DHA in humans and its metabolism were discussed. He also highlighted
that current research still largely focuses on practical biochemistry experiments and measurements
without specific prediction models. His feedback largely influenced our design of the efflux strategy
part and led to our final decision to put it under future plans.
Prof. Henry LAM
Our team was honored to reach out to Henry Hei Ning LAM who is a professor for the School of
Engineering at the Hong Kong University of Science and Technology (HKUST).
The consultation with Professor Henry helped us to enhance our Yeast Growth Model. He shared
with
us that tools like COBRA (Constraint-Based Reconstruction and Analysis) can already predict
the
basic growth and metabolic characteristics of Y. lipolytica, and these models could
be
used for
scaling up the process and integrating with a reactor model.
Regarding the modeling of promoter strength, Professor Henry suggested that we change the
rate
constants of the relevant reactions to simulate the impact of promoter strength.
Additionally,
he emphasized the need for wet lab data to calibrate and validate this approach. Professor
Henry
also advised us to start with a base model, and add our reactions but cautioned that this
approach may not be highly accurate, as we would not be able to optimize the reaction rates.
Incorporating the feedback from Professor Henry, we decided to explore the use of COBRA and
other
existing models to understand the basic growth characteristics of Y. lipolytica. We
will
also
decided to focus on developing a batch growth model, which will simulate the dynamics of
nutrients, biomass, and product formation.
We also took the approach of exploring the method of building our model from scratch.
Notably, we
decided on simplifying the model and focusing on a smaller set of reactions, rather than the
complexity of tools like COBRA. Additionally, we decided to seek out former iGEM students
recommended by Professor Henry to seek their guidance and support as we continued our
modeling
work.
By implementing these changes based on Professor Henry's feedback, we strived to develop a
more
robust and informed modeling approach for our project, leveraging both existing tools and
our
customized models.
Prof. Chao YE
We are honored to have NNU iGEM instructor——Professor Ye Chao to share his valuable experience
with us which are greatly beneficial to a clear direction of our team’s project.
Professor Ye Chao suggested several avenues for improvement, including the use of a MATLAB-based
COBRA toolbox in the genome-scale metabolic models, setting appropriate objective functions and
constraints, and exploring machine learning techniques for parameter estimation. He also gave
detailed suggestions of other types of models to consider including the ME model and the
Enzyme-constraint model.
The feedback received from Professor Ye Chao will be invaluable in refining and enhancing our
computational framework, ensuring that it provides a robust and versatile tool for the
scientific community.
Ms. Queenie LAM
To identify the pre-existing problems of DHA supplements and from a wide
perspective, omega-3 supplements in general, we approached a nutritionist, lecturer at HKBU
and
representative of the Hong Kong based nutrition company Negimenforall: Ms. Queenie Lam. By
touching upon Ms. Lam’s expertise in nutrition, we were able to further understand DHA
Deficiency’s prevalence in both Hong Kong and the global scale and of DHA’s nutritional
importance in building a healthy diet.
Ms. Lam described the DHA deficiency as a “silent killer”, emphasizing the lack of attention
to
the particular phenomenon. Within Hong Kong, she expressed how there was a rise in potential
industrial malpractice that may undermine the extraction yield and the health benefits DHA
supplements could bring citing this SCMP article. She also explained that the use of
genetically
modified organisms for DHA extraction was undesirable for consumers, pointing mainly to
unknown
long-term health effects as the deterrent.
Ms. Lam kindly published samples of our recipes onto the @negimenforall Instagram page,
through
which we were able to reach a wider audience.
Considering the cited concerns of consumers and the growing neglect of DHA in daily diets,
Ms.
Lam affirmed the potential benefits our project could bring to address these problems.
Ms. Queenie LAM
Prof Marshall LIU
We reached out to Professor Marshal Liu, an associate professor of engineering education at the Hong Kong
University of Science and Technology, regarding the scaling-up process of our two-stage reactor. He
provided us with invaluable insights, highlighting several critical factors to consider, such as oxygen
availability, pH levels, and temperature. In terms of real-world implementation, we also need to pay
attention to the costs, growth period, and biomass concentration of the chassis organism, Y.
lipolytica. Following our meeting, we gained a deeper understanding of the hardware components
of the bioreactor and made some minor changes to it. As of September, Professor Liu has taken on the
role of our supervisor, guiding us to conduct growth curve experiments using a 1-L bioreactor setup in
the food lab.
Dr Nicholson
Dr. Nicolson Siu is a neuropsychologist who has recently joined the Division of Social Science at the
Hong Kong University of Science and Technology as a lecturer. His expertise has been invaluable in
assisting us with the development of a market survey focused on the production of docosahexaenoic acid
(DHA) using yeast.
First meeting with Dr. Siu through Zoom
In our first meeting on 8th August through Zoom, we discussed several key aspects of the market survey
that will help us gather meaningful data.
A significant part of our discussion centered on gauging respondents' awareness of DHA. Instead of binary
yes/no questions, Dr. Siu proposed using a scale (1-10) to gather a spectrum of answers, rather than
saying to assess their degree of understanding. We discussed the necessity of asking questions that
differentiate between general awareness of omega-3 fatty acids and specific knowledge about DHA and we
decided to have questions like “Do you know about omega-3 fatty acids, like DHA and EPA”. Furthermore,
Dr. Siu emphasized the importance of consistency in terminology and clarity in questions. For example,
we deliberated on how to phrase questions about omega-3 fatty acids and DHA without leading respondents
or introducing bias, considering the effectiveness of a mini-quiz format to test awareness while
minimizing confusion. After the discussion, we decided to provide a general description for DHA so as to
let respondents have an objective view before answering the questions.
We also addressed the need to inquire about respondents' awareness of the environmental impact of DHA
production, framing the question to capture a range of opinions from negative to positive. Dietary
habits related to DHA-rich foods were another crucial topic; we agreed on the importance of including
questions that ensure respondents are not misled about which foods contain DHA. Additionally, we
discussed how to ask about the amount spent on DHA supplements, stressing the importance of consistent
pricing questions to facilitate data analysis. Dr. Siu suggested using the ratio of money spent on all
supplements to the money spent on DHA to conclude how likely people are to spend on DHA by proportion.
Lastly, we touched on the relevance of biosafety issues, particularly regarding GMO products, and how to
frame these questions without shifting the focus away from DHA itself.
Second meeting with Dr. Siu (left to right: Chloe, Dr. Siu, Henry, Jenny)
After modifying the survey based on the suggestions and advice given by Dr. Siu, we held another
face-to-face meeting on August 16. During this meeting, we went through the survey questions together,
to ensure the questions reduced as much bias as we could. One key topic of discussion was whether to
conduct street interviews for data collection and Dr. Siu recommended that we opt for focus groups.
Dr. Siu highlighted several advantages of using focus groups for our market survey. Focus groups allow
for a dynamic interaction among participants, providing a platform for diverse perspectives and
collective brainstorming. This group setting can generate richer discussions and insights, as
participants can build on each other's ideas and experiences. By engaging a small group of 6-10
participants, we can foster an environment that encourages open dialogue, which is particularly
beneficial for exploring complex topics such as awareness and understanding of DHA.
Overall, our discussions with Dr. Siu have provided critical insights into how to structure our market
survey effectively, ensuring that we gather meaningful data that aligns with our project objectives. His
expertise in neuropsychology and understanding of social science methodologies will greatly enhance the
quality of our research.
Dr. Aftab AMIN
We reached out to Dr. Aftab AMIN, who is a Teaching Associate at the Hong Kong University of Science and
Technology (HKUST). The meeting with Dr. Aftab AMIN primarily focused on incorporating Virtual Reality
(VR) as an educational tool within the summer program. The discussion centered around the potential of
using VR to enhance the learning experience, particularly in the field of biology and synthetic biology.
It was determined that human anatomy is one of the topics included in the summer program（we can
incorporate the link to our HP summer program here）, providing students with an opportunity to explore
the different parts of the human body interactively and engagingly. The rationale behind this decision
was to give students a chance to learn about biology through a VR-based approach, which could
potentially make the learning process more enjoyable and accessible.
We discussed how VR could be leveraged to create immersive and interactive experiences for students,
allowing them to visually observe and interact with the various components of the human body. This
approach was seen as a way to ease the learning process and make the overall experience more enjoyable
for the participants.
Through this outreach to Dr. Aftab AMIN, we gained valuable insights and perspectives on using VR as an
educational tool. The information gathered during the meeting was utilized to refine and improve the
summer program, thus ensuring that the integration of VR technologies enhances the learning experience
for the participating students.
We will continue to explore and implement strategies to effectively incorporate VR into the program, to
create a more engaging, interactive, and impactful learning environment for the students.
Utilizing VR headset as an educational tool
Dr. Mike CHEN
Left to right: David, Amanda, Robert, Chloe, Jenny, Henry, Dr. Chen
During our outreach to Dr. Mike Chen, the Manager of Business Development at the Office of Knowledge
Transfer of the Hong Kong University of Science and Technology (HKUST), we gained invaluable insights
into the entrepreneurial aspects of our project.
In our discussions, Dr. Chen emphasised the importance of understanding the competitive landscape,
particularly concerning DHA manufacturing. He highlighted HuaXi as a successful example that serves as a
benchmark for our aspirations. Dr. Chen also pointed out the challenges associated with the purification
process of DHA, which can complicate the production of our end product. His insights into the high
switching costs and the necessity for rigorous research in this area have been instrumental in shaping
our strategy.
Moreover, Dr. Chen encouraged us to clearly define our mission and vision, stressing the need for a
compelling narrative that illustrates our potential for success. He advised us on the importance of
establishing strong relationships with suppliers and manufacturers to ensure a stable supply chain and
cash flow, critical factors for any startup. While he suggested that a Business-to-Business (B2B)
model—such as selling our technique to manufacturers—might be more applicable to our project, he also
encouraged us to consider the potential for Business-to-Consumer (B2C) models to expand our customer
base and explore new market avenues.
Dr. Chen's perspective on the regulatory landscape surrounding genetically modified organisms and the
challenges of patenting technology has provided us with a clearer understanding of the hurdles we may
face. He recommended that we seek endorsements from leading professors in the field to enhance our
credibility and visibility.
In summary, Dr. Chen's assistance has been invaluable in helping us refine our business model and
navigate the entrepreneurial landscape of our project. His insights have clarified our path forward and
instilled a sense of confidence in our ability to succeed in this competitive industry.
Ms. Vivian Li
Introduction:
In our recent interview with Ms. Vivian from the Office of Knowledge Transfer (OKT), we gained valuable
insights into the patent application process and the commercialization of our project. The discussion
focused on the necessary criteria for patent eligibility, the resources required, and the challenges we
may face in transitioning from research to market.
Patent Application Criteria:
Ms. Vivian outlined the criteria for applying for a patent, emphasizing the need for our project to
demonstrate novelty and usefulness. She highlighted that specific elements of our research, such as the
yeast strain, the metabolic pathway, and the optimized recipe, could be candidates for patenting.
Particularly, the introduction of genetic components into the yeast and the method of producing DHA
(docosahexaenoic acid) were identified as key areas to consider for patent protection.
Application Procedure and Documentation:
The patent application process involves several steps, and Ms. Vivian provided clarity on the general
procedure and timeline. She explained that we would need to prepare specific documents, including the
university's invention disclosure form and a manuscript. The university would handle the remaining
aspects of the application process, which is crucial for protecting our intellectual property before any
public disclosure.
Transitioning to Commercialization:
One of the significant challenges discussed was the transition from research to commercialization in
biomanufacturing. Ms. Vivian pointed out potential hurdles such as quality control, regulatory
compliance, and the costs associated with bringing our product to market. She suggested that a B2B
(business-to-business) model might be more effective for our project, targeting raw material suppliers
and manufacturers that use DHA, rather than pursuing a B2C (business-to-consumer) approach.
Marketing Strategies:
When discussing marketing strategies, Ms. Vivian advised us to focus on the viability of DHA and its
applications in the health sector. She emphasized the importance of highlighting the benefits of our
product to strengthen our marketing efforts. If we were to consider a B2C approach, she suggested
exploring how to package our end products effectively to attract consumer attention.
Collaboration and Industry Engagement:
Ms. Vivian also stressed the value of collaborating with industry partners to scale our production. She
recommended identifying key players in the DHA manufacturing space and exploring potential partnerships.
This collaboration could facilitate a smoother transition from research to market and help us navigate
the complexities of commercialization.
Conclusion:
The insights gained from our interview with Ms. Vivian have clarified several critical aspects of our
project, particularly regarding patent applications and commercialization strategies. As we move
forward, we will focus on refining our approach to patenting and developing a robust business model that
aligns with the recommendations provided. This strategic planning will be essential for ensuring the
success of our project and its potential impact in the market.
Prof. Coral PUIG
Professor Coral Puig Garrigo combines her expertise in evolutionary anthropology, neuro-marketing and
consumer psychology to help businesses navigate the ever-evolving landscape of market trends. In
addition to her work in the private sector, Coral serves as an Adjunct Marketing Professor at HKUST's
Business School.
We recently consulted with Professor Puig regarding our project storyline, and her insights have been
invaluable in refining our plot. Initially, we focused on the market for DHA and discussed the
disadvantages of current production methods, such as fishing and microalgae cultivation, before
introducing our proposed solution. However, after our discussion with Professor Puig, we made several
significant adjustments to our narrative to enhance clarity and engagement.
Professor Puig suggested that we begin by highlighting the importance of DHA, emphasising its positive
effects on brain health. By framing our project around the idea that DHA can enhance intelligence, we
aim to capture the audience's attention more effectively. She advised us to avoid overly complex or
technical explanations, ensuring our message is accessible and compelling.
We have also decided to present our project first, followed by a discussion of the limitations of current
DHA production methods.
Overall, we are incredibly grateful for Professor Puig’s feedback, which has already proven instrumental
in shaping our storyline.
Researcher WANG
Meeting with Mr.Wang Dapeng with BNUZH-China and UM-Macau teams
Mr. Wang Dapeng is an associate researcher at the China Popular Science Research Institute and a director
of the China Popular Science Writers Association. He specialises in the following fields: science
communication theory and practice, social media science communication, scientist-media relations, etc.
He has undertaken dozens of research projects in recent years. In recent years, he has undertaken dozens
of scientific research projects; published more than 300 commentaries in mainstream media such as
《中国科学报》,《光明日报》,《科技日报》,《人民日报》etc., and set up columns in Guangming.com and Science Times; published
dozens of papers in various core journals; translated and published a number of monographs on the theory
of popularisation of science (two translations have won the awards in the industry), and published a
monograph on the theory of popularisation of science, entitled “The Vision and the Gateway”, which is a
book of 40 popular science writers. He has also published the monographs of science popularisation
theory and research, 《愿景与门道：40位科普人的心语》,《谈科与论普-科普人“出圈”手册》, and so on. He has conducted more than 100
popularisation training sessions for various types of scientists and technologists.
On 12th September, we engaged in a productive discussion with Mr. Wang Dapeng regarding effective science
popularisation strategies, particularly in collaboration with the BNUZH-China and UM-Macau teams.
During our meeting, Mr. Wang shared his insights on various facets of science communication, emphasising
the power of multimedia tools such as videos and concise posts to effectively popularise the field of
synthetic biology. He urged us to consider the emotional dimensions of our messaging, highlighting the
importance of connecting with our audience's perspectives and concerns. Storytelling emerged as a
pivotal technique in making complex scientific concepts more accessible and relatable to the general
public.
In the context of our project, which focuses on utilising yeast to produce DHA (Docosahexaenoic Acid),
Mr. Wang pointed out the necessity of addressing ethical considerations associated with yeast usage. It
is vital to assess any potential negative effects of yeast on human health, as well as its overall
safety. He stressed that while it is essential to provide comprehensive information, excessive focus on
the drawbacks may inadvertently instil fear among the public. Consequently, in our market research, we
should aim to strike a balance by not overly emphasising the cons of DHA.
Mr. Wang also commended our science communication efforts, reinforcing the notion that successful
engagement is not solely about conveying facts, including primary school workshops, high-school
workshops etc. Rather, it involves establishing a connection with the audience, addressing their
apprehensions, and offering practical solutions to their concerns.
In conclusion, our dialogue with Mr. Wang underscored the multifaceted nature of effective science
communication. By leveraging emotional resonance and storytelling, we can enhance our outreach efforts
and foster a deeper understanding of scientific advancements within the community.
Prof. Ren Jian
To deepen our understanding towards the project application, we co-interviewed Professor Ren Jian (任间),
senior biotech entrepreneur and PI of Sun Yat-sen University iGEM since 2012, together with members from
UM_Macau and BNUZH teams.
Our discussions centered on the project's commercialization and technical challenges. Key components for
successful commercialization, including core resources, products, value propositions, and target
customer segments, were highlighted. Prof. Ren emphasized on articulating these aspects clearly from a
technical standpoint, showcasing advantages over traditional methods and the competitive strengths, for
example cost-effectiveness and purity, with quantifiable metrics. Regarding our project, stability and
cost considerations for future mass production were noted, with stability as a potential highlight. Also
AlphaFold was recommended for modeling.
From his perspective, laboratory tests serve more as a platform to demonstrate feasibility. He also
touched upon the complexities of synthetic biology entrepreneurship, emphasizing the need for stringent
product qualifications and adherence to industry standards, particularly in food-grade applications. The
meeting underscored the importance of combining technical expertise with attractive design and sound
business logic, ensuring a robust and marketable project.
Prof. Karl TSIM
We reached out to Professor Karl Tsim, who is the Chair Professor in the Division of Life
Science
at the Hong Kong University of Science and Technology (HKUST). We were able to receive
valuable
insight into technical challenges, information on potential investors, and regulations to be
aware of.
Professor Tsim shared his experiences working with a local Hong Kong-based company that
isolated
DHA from fish meat. He affirmed the well-established health advantages of DHA for brain
development, cardiovascular health, and stroke prevention.
In terms of the technical challenges, Professor Tsim highlighted that the enzymatic pathways
involved in DHA production are quite complex, making recombination in yeast a significant
hurdle. He pointed out that we would need to thoroughly investigate whether yeast can truly
outperform other approaches before pursuing that route. Additionally, we would need to focus
on
efficient purification of the DHA extract.
To attract potential investors down the line, the professor suggested we explore
opportunities
for intellectual property protection. Investors would also be interested in the
investment-to-profit ratio and timelines. Differentiating factors could include optimizing
the
ratio of DHA compounds produced that offer us advantages.
On the business side, Professor Tsim advised us to reach out to potential manufacturers in
mainland China that utilize DHA as a raw material for their products, as the Hong Kong
market
currently lacks major DHA suppliers.
Regarding regulations, the professor indicated that there are relatively few restrictions on
DHA
in Hong Kong, primarily requiring heavy metal testing and ensuring the product does not
contain
Western medicines like aspirin.
Overall, the discussion with Professor Tsim provided valuable guidance to refine the
project's
scope, technical priorities, and potential commercialization pathways. Our team decided to
follow the advice, including the project direction.
Algreen
From left to right, First row: Dhruv, Tasin and David; Second row: Henry, Jenny
and Chloe
During the meeting with
AIGreen
, we gained valuable insights into the microalgae industry that will
significantly influence the direction of our project. While AlGreen’s expertise in microalgae
cultivation and lipid extraction was impressive, our focus was on how we can apply their knowledge to
enhance our own initiatives.
The meeting first highlighted the challenges associated with microalgae cultivation, such as preventing
contamination and the necessity for tailored nutrient media for different species. Tasin and Dhruv, the
senior research analyst and the founder of AlGreen, shared their experiences with algae cultivation and
emphasised the importance of adhering to established protocols. AlGreen stressed the role of bioreactors
in providing precise control over cultivation parameters, which is critical for successful microalgae
production. This insight has prompted us to prioritise the development of robust protocols for the
cultivation, ensuring high quality mass is maintained.
We also asked about the design and cost considerations of bioreactors for algae cultivation. Tasin
explained the essential features of these reactors, including constant airflow and controlled lighting,
while addressing the challenges of sterilising larger chambers. We then considered these factors in our
cost analysis for scaling up production, specifically evaluating the differences between bubble and
tubular reactors and their respective efficiencies in various environments.
In terms of biomass and lipid extraction strategies, Dhruv and Tasin emphasised the importance of
concentrating on microalgae biomass rather than getting caught up in complex culturing and nutrient
hardware. This advice has led us to refine our project scope, allowing us to allocate resources more
effectively towards efficient lipid extraction methods. We are exploring patented techniques shared by
AlGreen that could streamline our extraction process.
The conversation then covered the challenges of extracting lipids from algae. Tasin explained that while
genetically modifying the cell wall to facilitate extraction is a complex task, they are focusing on
physical extraction methods like hydrothermal liquefaction and sonication, which are scalable and
require less heat and pressure. The team discussed the energy-intensive nature of the extraction process
and explored ways to minimise electricity costs, such as utilising natural sunlight and solar-powered
air pumps.
Additionally, we discussed the strategies for producing fatty acids using yeast, noting that it may be
more efficient to keep fatty acids within the yeast cells for easier extraction. Dhruv shared their
company's approach of scaling down by selling indoor algae air purifiers to households, which would help
reduce carbon emissions while generating a profitable income stream.
AlGreen's commitment to sustainability aligns perfectly with our project goals. They suggested we take a
look at Life Cycle Assessment (LCA) methodologies, evaluating the carbon footprint of our DHA production
process. We planned to evaluate the carbon footprint of our DHA production process, identifying areas
for improvement that will align with market demands while positively impacting the environment.
Overall, the meeting helped us understand the challenges faced by microalgae cultivation, fostering
dialogue about the potential of microalgae and biotechnology in addressing nutritional and environmental
challenges. The insights gained helped us make DHA production from yeast a more sustainable solution.
Dr. Zhu Yuanmin
Recommended by Prof. Yu Longjiang (余龙江), we consulted with Dr. Zhu Yuanmin (朱圆敏) from Huazhong University
of Science and Technology, a experienced patent owner for DHA-producing Schizochytrium sp..
During the meeting with Dr. Zhu, various critical factors for industrial production and optimization were
deliberated. Key considerations included the focus on enhancing DHA production capabilities while
maintaining the quality, and adherence to national regulations for food production. She shared that
their competitiveness in engineering was achieved through directed mutagenesis to avoid the strict
regulatory approval of new commercial strains, which can take years, underlying one of the biggest
challenges faced in the application of synthetic biology to supplement or food industries. The usage of
approved strains was highlighted as crucial, particularly to ensure consistency in DHA percentage in
total fatty acids, which is under regulation of existing quality standards. The feedback implies the
necessity to include a future strategy to inhibit native fatty acid synthesis and redirect resources to
DHA production. Dr. Zhu also suggested improving the compatibility of our system for wider industrial
uses in the future.
Moreover, our discussion spanned to avoiding contamination of carbon sources by using glucose, and
employing strategies for metabolic optimization through modeling, as performed in our project.
Challenges related to GMO concerns, industrial scaling, and market adoption of DHA products were also
brought to light. We learned from the dialogue that utilizing existing facilities through collaboration
would be more cost-effective and thus recommended. She also provided us with insights into patenting,
and proposed various modes of collaboration including one-way patent transfer or developing iterations
of the engineered strain through a long-term contract, highlighting the need for a well-thought
implementation plan.
ALGDEHA
ALGDEHA
is a Turkish startup company focused on omega-3 fatty acid production from microalgae. We luckily
connected to Mr. Mehmet Can Sucu, founder of ALGDEHA through Linkedin. We had a fruitful conversation,
deepening our understanding towards the current industry and entrepreneurship.
Mr. Susu shared that ALGDEHA aimed to replace fish meal and fish oil in fish feed formulas to compete on
price and content. Historical data indicated that fish-based products were priced at $4,000 per ton,
while the current leading producer Veramaris produced microbial-based oil at $8,000 per ton, hinting at
a production cost range of $4,000 to $8,000 per ton. Therefore, to succeed, matching or surpassing
Veramaris's productivity levels is crucial, emphasizing the need for high productivity and DHA content.
Calculations and literature suggested that achieving 100 g/L of dry Schizochytrium sp. biomass is
possible, akin to Veramaris's production levels. A similar analysis can be carried out with our strain
Yarrowia lipolytica.
Mr. Susu pointed out that the primary focus remains on optimizing fermentation conditions for peak
productivity, with factors like glucose concentration, nitrogen depletion, and a two-step aeration
process critical for maximizing productivity and lipid accumulation. It indicates the importance of
implementing a stage control strategy in our project. Beyond optimization, according to him, genetic
manipulation and synthetic biology approaches are vital to overcome productivity limits, for example
transferring it to a more competitive organism. However most of them remain as a future direction. This
information coincides with our idea to introduce the PUFA synthase pathway into Yarrowia
lipolytica,
indicating its industrial significance.
For entrepreneurship, we learned that effective communication of the startup's story and milestones to
investors poses challenges due to the significant capital investment and long-term commitment required
for mass-scale operation, usually between 100,000 and 300,000 liters. Demonstrating enhanced
productivity and lipid accumulation at the lab scale and highlighting a path to industrialization with
lower capital investment through contract-based production trials were suggested as key strategies to
attract investor interest and ensure long-term viability.
EFSA
In our ongoing efforts to enhance our understanding of the regulatory landscape surrounding Genetically
Modified Organisms (GMOs), we reached out to the European Food Safety Authority (EFSA) with specific
questions related to our project on yeast-derived DHA. Although we were unable to secure an interview,
EFSA provided valuable insights that have significantly influenced our project development.
EFSA's primary function is to evaluate the safety of GMO products before market authorization decisions
are made by risk managers. This evaluation focuses on potential impacts on human health, animal health,
and the environment, highly aligns with our project. The communication challenges regarding GMOs and
public perceptions are largely managed by the European Commission and national authorities, prompting us
to consider more effective communication strategies in our outreach efforts regarding yeast-derived DHA.
EFSA emphasised the importance of adhering to their guidance documents for safety assessments of novel
foods, leading us to incorporate this understanding into our project by ensuring that our safety
assessment protocols align with EFSA's methodologies.
Based on the insights gained from EFSA, we have made several modifications to our yeast-derived DHA
project. We revised our safety assessment protocols to align with EFSA's guidance documents, ensuring a
comprehensive evaluation of potential health impacts. We will develop a communication strategy that
addresses public perceptions of GMOs, aiming to foster transparency and trust among stakeholders.
The outreach to EFSA has proven invaluable in refining our approach to developing yeast-derived DHA,
enhancing our understanding of regulatory requirements and guiding us in adopting best practices for
safety assessments and communication. We are committed to leveraging this knowledge as we advance our
project and contribute positively to the industry.
Participant Demographics
The survey initially collected responses from 126 participants, aiming to understand the perspectives of
individuals from different nationalities and age groups regarding DHA.
Over half of the respondents are from Hong Kong, we received responses from a total of 11
nations/regions, with participants from China and India comprising 19% and 13% of the total,
respectively. This help us have a better understanding for both local and global market in some way.
We gathered a wide range of respondents aged between 14 and 60, which enhanced the conclusiveness of our
results by allowing us to consider a broader audience.
Most respondents hold a university-level education.
Dietary Preferences
From the responses, we found that 86% of participants prefer to eat fish and fish-based products,
indicating that they may have a fair intake of DHA from these sources compared to those who dislike fish
or cannot consume it.
Awareness of Omega-3 Fatty Acids
Less than 70% of respondents reported awareness of omega-3 fatty acids, including DHA and EPA. This
underscores the necessity for us to enhance outreach efforts to raise awareness about the benefits of
DHA and its importance.
In our discussions with Professor Nicolson from SIU, we received guidance on how to make the survey more
reader-friendly. To achieve this, we focused on maintaining consistent terminology, using the broad term
"omega-3 fatty acids" alongside specific examples like DHA.
Importance of DHA Awareness
Nearly half of the respondents rated their awareness of DHA as 4, and nearly 20% rated it a 5. This
suggests that a majority of the respondents recognize the importance of DHA for health.
However, despite this relatively high awareness, half of the respondents do not take any omega-3 or fish
oil supplements, and 33% have only taken them once. This indicates that while the audience may be aware
of omega-3 and DHA, few actively incorporate them into their diets through supplements. This suggests a
potential market gap that our projects aim at narrowing this existing gap.
Environmental Concerns
On a positive note, 22% of respondents expressed no concern regarding the environmental impact of fish
oil production. While most respondents (24%) remained neutral, 20% rated their concern at 4 or above,
indicating a significant level of concern that should not be overlooked. This highlights the need for
our team to focus on biosafety and public education to clarify these concepts and alleviate concerns,
also explaining why we are using yeast to make DHA particularly and how that could help ease the
environmental burden.
Willingness to Try DHA Products
A majority of respondents indicated a willingness to try DHA products, rating their interest at 4 or
above. This reflects an optimistic outlook and an open-minded attitude toward new DHA products in the
market, suggesting a commercial opportunity. However, 33% remained neutral, indicating that their
decision may depend on market responses regarding the effectiveness and safety of the product.
Key Considerations for DHA Products
Price, safety, and health benefits emerged as the top three priorities, with over 90% of respondents
selecting these factors as crucial considerations. Surprisingly, brand reputation ranked highly, with
nearly 70% of respondents prioritizing it over environmental impact (58%) and source (58%). This
suggests we should highlight these top three priorities in our collaborations with businesses that have
strong brand images. The lower emphasis on sourcing indicates that respondents may be open to our
project utilizing yeast as a source.
Safety Concerns
Surprisingly, 55% of respondents rated their safety concerns as 3 or below, indicating they are less
worried about safety. Additionally, 23% remained neutral, while 21% expressed more concern. This
suggests a need for ongoing education to enhance awareness regarding the safety aspects of our project,
which we will address through related outreach.
Respondents’ Perspectives on Safety
To better understand respondents’ concerns, we included open-ended questions and received a range of
comments. Professor Nicolson advised that we avoid leading assumptions about DHA's effectiveness to
generate more objective results based on participants' original thoughts.
Concerns expressed by respondents:
- (Uncertainty about Product Safety) Uncertainty about the testing process
- Unknown long-term effects of the product
- (Familiarity and Knowledge Gaps) Lack of familiarity with such products
- (Health Risk Aversion) Reluctance to take health risks
- (Perceived Importance of Alternatives) Perception that other vitamins and proteins are more
important than fish oil supplements
Reasons respondents expressed less concern:
- (Trust in Science and Technology) Trust in GMO products/science Confidence in the product having
undergone a trustworthy quality control process
- (Familiarity with Yeast) Familiarity with yeast from the brewing industry
- (Quality Assurance) Assurance that the product meets standards like Good Manufacturing Practice
(GMP)/FDA approval
- (Established Reputation and Evidence of Safety) Established brand reputation Belief that sufficient
research and experiments have been conducted
- The perception that it is a better replacement for fish
- Prior knowledge in biochemistry/biotechnology
- Acceptance of GM technology as a developed method
- Awareness of existing GM products in the market
- The belief that benefits outweigh risks
- Lack of significant negative effects from GM products thus far
Key Consideration
- Points Confirmation that official safety standards have been met
- Assessment of whether sufficient information and knowledge have been communicated and understood
Preferred Methods of DHA Intake
From the survey, we found that 67% of respondents prefer cooking as their method of DHA intake, while
nearly 30% are open to trying supplements. Our project aims to produce DHA sustainably while considering
these preferences helping people who would like to cook such as the design of DHA recipe. This question
helped enhance our understanding of the public's approach to DHA intake.
Reasons for Cooking vs. Taking Supplements
Reasons for preferring cooking:
- (Sufficient Dietary Intake)The belief that dietary intake is sufficient
- (Preference for Organic Approaches)Preference for a more organic approach
- Perception of being more natural and healthy
- (Simplicity and Enjoyment)Simplicity and ease of preparation
- Enjoyment of taste and texture
- Trust in food over supplements
- (Cost-Effectiveness)Lower cost, as meals are seen as necessary while supplements are viewed as
extras
Reasons for preferring supplements:
- Convenience
- (nutrient concentration) Higher nutrient concentration
- Ease of consumption
- (Time constraints)/ (social influence)Lack of time or equipment for cooking, especially among
students Influence from social trends
