Human Practices & Integrated Human Practices

Human Practices

Overview

Excessive consumption of high-calorie sugars poses risks of obesity, diabetes, hypertension, and other diseases. Consequently, an increasing number of individuals opt for low-sugar and low-fat food products. Inspired by reports categorizing aspartame as a Group B carcinogen, our team conducted market research by visiting supermarkets and local markets. Observing a growing trend in the use of natural sugar substitutes such as erythritol or xylitol replacing glucose or sucrose in various food products, we recognized the challenges in the natural extraction and semi-chemical synthesis of these substitutes. Motivated to address these challenges, we aimed to leverage synthetic biology techniques to engineer lipid-producing strains of Yarrowia lipolytica for de novo synthesis of erythritol and xylitol. In the critical experimental design phase, we collaborated with experts in the field to formulate appropriate experimental plans. We conducted online surveys and public education initiatives in synthetic biology to explore public awareness and acceptance levels regarding sugar substitutes. Meanwhile, we conducted a Women in STEM (Science, Technology, Engineering, and Mathematics) themed outreach activity, aiming to promote active female participation in scientific research endeavors. We dedicated our efforts to this cause. The objective is to advance the development of a sustainable and bio-based alternative to traditional sweeteners, promoting both health-conscious consumer choices and the application of synthetic biology in the food industry.

In the whole HP activities, each part is complementary to each other, forming an organic whole.

Silver

In the design of our HP project, each segment of activities is driven by a specific goal, and we have achieved the anticipated outcomes. We leverage the results obtained through HP to refine and promote our project, striving to meet our desired objectives. As the project unfolds, we have categorized the HP goals into five distinct sections based on task types: Background, Project Research, Project Design, Project Promotion, and Project Optimization and Improvement. These categories serve as a guiding framework for our activities. Furthermore, to ensure the standardization and ethical adherence of our HP initiatives, we have sought advice from relevant experts to guide the design of our activities.

Background

1.Sugar Intake: Overview and Far-reaching Impact on Health

Learning from recent news reports, we've come to understand the prevalent and persistent issue of diabetes in our country. The long-term consumption of high sugar levels not only leads to diabetes but is also associated with the rise of other health complications. What's particularly worrisome is the increasing trend of diabetes affecting individuals at younger ages, indicating a closely intertwined relationship between dietary habits and health. This observation has sparked a thoughtful reflection among the members of us project about the importance of a healthy diet. The erratic lifestyle of young people, coupled with a strong preference for high-sugar and high-calorie foods, is undeniably impacting our overall health. As a result, members of our ZJFH-Nanjing team have decided to shift our focus to address the sugar content in diets. Our efforts are now directed towards the development of alternatives to sugar in food, recognizing the urgency of promoting healthier eating habits.

2. Research on application of sugar substitute: Visit supermarket

On April 23, 2023, our members of our ZJFH-Nanjing team conducted a market survey of sugar-free beverages at SuGuo supermarket. The market for sugar-free beverages has been growing steadily, primarily encompassing sugar-free tea drinks and sugar-free carbonated beverages, with sugar-free carbonated drinks dominating the market. Currently, most of the food and beverages containing sugar substitutes on the market primarily use erythritol, with a smaller amount of xylitol. Some brands may also offer both sugar substitute-added products and products that use regular sugar. Products with sugar substitutes are generally priced higher than those using regular sugar, and this price difference is often attributed to the higher extraction cost of sugar substitutes.

Simultaneously, we randomly interviewed several college students, and they expressed a preference for sugar-free beverages. Due to concerns over aspartame, which is classified as a Group B carcinogen, erythritol is gaining more trust. With the increasing awareness of health consciousness, we believe that the market for the application of sugar substitutes will continue to expand.

Project Research

1. Research on application of sugar substitute: Visit Nanjing Center for Disease Control and Prevention

Introduction: Pei Xinyi, from Nutrition and Food Hygiene at Nanjing Center for Disease Control and Prevention

On May 10, 2023, members of our ZJFH-Nanjing team visited the Nutrition and Food Hygiene at Nanjing Center for Disease Control and Prevention, where we had the opportunity to meet with teacher Pei Xinyi. Teacher Pei emphasized that excessive consumption of sugary foods can lead to obesity and increase the incidence of diseases such as diabetes, blood clots, and cardiovascular diseases. While she noted that sugar substitutes in food cannot be relied upon as a long-term preventive measure for these diseases, they can play a certain preventive role when consumed in moderation. Teacher Pei recommended limiting the daily intake of sugar substitutes to around 4 mg.

Furthermore, Teacher Pei urged everyone to maintain regular physical activity and cultivate healthy lifestyle habits. This advice underscores the importance of comprehensive health management, aiming to reduce the risk of chronic diseases associated with a high-sugar diet. It also highlights the significance of exercise and good lifestyle habits in promoting overall health.

2. Research on application of sugar substitute: Visit Nanjing University of Chinese Medicine

Introduction: Teacher Wang Qingzhuo, from the School of Integrative Medicine at Nanjing University of Chinese Medicine

On May 16, 2023, members of our ZJFH-Nanjing team visited the School of Integrative Medicine at Nanjing University of Chinese Medicine and had the privilege of meeting with teacher Wang Qingzhuo. Teacher Wang provided us with insights into the role and future applications of sugar substitutes in diet. Subsequently, he elucidated the mechanisms by which excessive sugar intake leads to diseases and discussed the current primary treatment modalities. Teacher Wang also emphasized the importance of strict dietary control for individuals with diabetes. He suggested that these individuals can consider incorporating foods with added sugar substitutes in moderation.

This approach not only satisfies the patients' cravings for sweetness but also minimizes the impact on elevated blood sugar levels. Teacher Wang's advice underscores the potential benefits of sugar substitutes in managing the dietary needs of diabetes patients.

Project Design

1. Interview Teacher Wang Qingzhuo

Introduction: Teacher Wang Qingzhuo, from the School of Integrative Medicine at Nanjing University of Chinese Medicine. The research field is to transform non-model microorganisms based on synthetic biology technology to directionally produce biochemicals.

After the previous market research and expert discussions, we have roughly defined our research goals, aiming to achieve targeted microbial synthesis of erythritol and xylitol. To refine our experimental design, we engaged in in-depth discussions with teacher Wang Qingzhuo. He analyzed various industrially advantageous strains and identified the most suitable microbial chassis strains for our target products (erythritol and xylitol). Additionally, we explored suitable gene editing technologies for these strains, laying the foundation for the determination of our subsequent experimental plans.

In conclusion, teacher Wang Qingzhuo expressed high expectations for our project and welcomed us to consult him at any time.

2. Interview Teacher Chen Yuan

In order to refine the design of the project content and provide reliable guidance for experimental design, we conducted multiple interviews with expert professors in the relevant field. Teacher Chen Yuan suggested the utilization of erythritol-inducible promoters for the construction of the screening system. Professor Xue Feng recommended exploring enzymes from different sources in the synthetic pathway to enhance the production of erythritol.

3. Communication with Nanjing Normal University

Introduction: Associate Professor Shi Tianqiong, from the School of Food and Pharmaceutical Engineering at Nanjing Normal University. His research focuses on the development of CRISPR/Cas9 genome editing technology toolbox in filamentous fungi and related studies on reshaping complex metabolic networks in microorganisms using synthetic biology techniques.

On June 20, 2023, our members of our ZJFH-Nanjing team visited Associate Professor Shi Tianqiong at the School of Food and Pharmaceutical Engineering, Nanjing Normal University. He led us on a tour of the synthetic biology laboratory, providing insights into laboratory safety protocols and demonstrating the proper use of various instruments. Our senior colleagues demonstrated standard experimental procedures, expanding our knowledge and enhancing our understanding of laboratory safety and the ethical considerations of biological modifications. This visit contributed to our academic growth.

In the final part of our visit, we discussed our experimental design with Professor Shi, seeking his advice on aspects such as the selection of luciferase and gene editing tools. Associate Professor Shi expressed his hope for the successful completion of our project under safe conditions.

Project Promotion

1. Development and Promotion of a Science Outreach Public Account

The results of Public Engagement have indicated that the general public often struggles to sugar intake, distinction between natural and synthetic sugar substitutes and sweeteners. As a response to this observation, our team has taken a step further by developing a science outreach public account called "Cellular factories for sugar substitutes" (Fig. 11). In this science outreach platform, we aim to educate the public not only about the potential dangers of excessive sugar consumption but also about the distinctions between natural and synthetic sugar substitutes. Additionally, we provide insights into the ongoing projects within our team.

Through "Cellular factories for sugar substitutes" we are committed to raising awareness and clarifying misconceptions related to sugar and its alternatives. Our goal is to empower the public with accurate information and encourage healthier dietary choices while shedding light on our team's current research endeavors.

2. Field Trips

Due to the limitations of online promotion, we opted to conduct outreach activities at a densely populated subway station over the weekend to ensure a wide reach (Fig. 12). Additionally, we visited communities to provide educational talks to families, with a focus on adults around 40 years old, children around 10 years old, and seniors around 60 years old, ensuring a diverse and representative audience. Through these interactions, we found that some parents were already aware of the potential harm caused by excessive sugar intake and actively controlled their children's sugar consumption, especially for those in the developmental stage.

Furthermore, we observed that older individuals expressed concerns not only about the risks associated with excessive sugar intake leading to conditions like hypertension and diabetes but also about the safety of consuming natural sugar substitutes.

In addition to community visits, we organized outreach activities in shopping malls to address public concerns regarding the safety and intake levels of natural sugar substitutes. The response to these events was highly positive, with many attendees gaining a preliminary understanding of sugar substitutes for the first time. These offline activities proved effective in eliminating concerns and increasing public awareness of the role and safety of natural sugar substitutes.

3. Organization of the Women in STEM theme with several teams

In a recent group meeting, we discovered that some students had experienced advice from parents or relatives, suggesting that as girls, they might have an advantage in studying humanities. However, these students were more inclined toward the sciences. These students are passionate about the fascinating reactions in physics, chemistry and biology experiments, and they enjoyed deciphering the logical relationships within formulas. Unfortunately, they often found themselves compelled to choose subjects they didn't particularly enjoy due to the stereotypical impressions of their parents.

Similar experiences were shared by female members in our team. They had been recommended by friends or teachers to pursue humanities, but their passion for biology, physics, and chemistry remained strong. In their experiments, they displayed meticulous attention and enthusiasm, contrary to societal perceptions that women are not suited for or proficient in the sciences. While these team members were fortunate enough to persevere in studying subjects they loved, we are aware that many girls are uncertain about whether they truly enjoy humanities and whether they have the confidence to achieve success in science disciplines that they love. Moreover, the internal struggle about whether to follow their passion for the sciences persists.

To rekindle confidence in girls regarding STEM (Science, Technology, Engineering, and Mathematics) subjects, we decided to collaborate with BS_United, DUT_China, AIS-China, Shanghai-MedX and RDFZ-CHINA to organize a "Women in STEM" themed event. Following the event, we also aimed to establish a platform for sharing educational resources that inspire and support women.

The Women in STEM theme activities were discussed and researched by BS-United-China, DUT-China, AIS-China, Shanghai-MedX, and RDFZ-CHINA. ZJFH-Nanjing shared the efforts and assistance provided by women in the early stages and experiments of the project, focusing on outstanding female scientists in the research of sugar substitutes and related fields. The event took place in an online format, with a maximum of 103 participants in the virtual meeting room.

1）What the panel shared？

BS-United-China: Girls can't learn mathematics, physics and chemistry well? Tear off the label theme of subject prejudice.

DUT-China: Don't extinguish the love for STEM because of gender.

AIS-China: From Hair to Scalp: The Secret of Male and Female Hair Loss, Attitude Differences and Social Dilemma Exposed.

Shanghai-MedX: Colorectal Cancer Drug Screening System, and Female Scientists in the Cancer World.

RDFZ-CHINA: Don't Let Acne affects women's self-confidence and leads to appearance anxiety.

ZJFH-Nanjing: Sweet through the sugar control period . What did those female scientists do?

2）Establishment of a Resource Sharing Platform

After the themed lectures, ZJFH-Nanjing, along with other teams consolidated the data, readings, literature, presentations, videos, and other materials beneficial for women to choose their majors based on their conditions and interests. This information was shared through email communication, and each team's presentation slides from the meeting were uploaded to a cloud drive. This allowed teams to extract needed information and materials from one another. Various forms of promotional lectures, exhibitions, and activities were organized in school clubs, communities, and other locations.

Project optimization and improvement

1. Consult with relevant experts to improve project design

To explore the feasibility of synthesizing sugar substitutes through microbial processes, we engaged in an online exchange with Associate Professor Xu Yameng from the Institute of Cotton Research of CAAS. Professor Xu shared insights into the current main production pathways of certain sugar substitutes. Sugar substitutes are primarily derived through natural plant extraction and semi-chemical synthesis. However, both methods face limitations. Natural sugar substitutes have low concentrations in plants, and the extraction or synthesis costs are high, making it challenging to meet the demands of large-scale production in factories.

Associate Professor Xu highlighted the vast potential of synthetic biology in addressing these challenges. She suggested that utilizing synthetic biology approaches might reduce the production costs of sugar substitutes. Currently, she is using synthetic biology methods to synthesize another type of natural sugar substitute and has achieved some promising results. Regarding our experimental project, Associate Professor Xu provided valuable advice. She suggested that we explore suitable target genes from the literature, construct plasmids accordingly, and then express them in host cells to produce sugar substitutes.

2. The lecture provided us with the new options of sugar substitutes

Introduction: Professor Osman, from Adana Alparslan Türkeş Science and Technology University in Turkey, has been dedicated to research in food processing technology for many years. He is internationally recognized as an expert in the field of stevia, having published over 70 academic journal articles and conference papers in related areas.

On September 28, 2023, Professor Osman delivered an insightful academic lecture titled "STEVIA: The Alternative of Sugar" to our team and the faculty and students of Nanjing Normal University. During the lecture, Professor Osman provided an overview of the stevia industry in China, Turkey, Paraguay, and other countries, covering cultivation, processing, and import-export trade. He also offered insights into the prospects of the sweetener industry, with a particular emphasis on the application scope and significance of sugar substitutes in the current food sector. Professor Osman's lecture shed light on the global landscape of stevia production and its potential impact on the food industry.

After the insightful lecture, we had a valuable exchange with Professor Osman. He not only praised our project design and content but also offered constructive insights. Particularly, he emphasized the potential of leveraging synthetic biology techniques to enhance the microbial production capabilities of xylitol and erythritol. We discussed with Professor Osman the previously suggested ideas of selecting a screening system and using synthetic enzymes from various sources. His enthusiastic endorsement of this strategy bolstered our confidence in achieving a significant improvement in the yield of these two products.

3. Looking for the commercial future of the project

Nanjing Zhihe Biotech: Exploring the Commercial Future of Sugar Substitutes in Oil and Terpene Compound Production

Nanjing Zhihe Biotech is a bio-manufacturer specializing in the synthesis of oils and terpene chemicals. Leveraging synthetic biology and advanced manufacturing technologies, the company produces high-value chemicals through oil-producing microorganisms such as oleaginous fungi and lipid-producing yeasts. Committed to providing "green, innovative, value-added" solutions in the realm of lipid chemistry, the company aims to make significant contributions to society.

The company acknowledges the current predominant use of sugar substitutes in the food industry but poses an intriguing question: Is there potential to rapidly apply sugar substitutes in food additives while concurrently co-producing oil-based compounds or terpene compounds? This exploration suggests a new commercial horizon for sugar substitutes in uncharted territories. The company expresses its interest in further collaborative discussions after stable production is demonstrated. This strategic consideration demonstrates a keen insight into the future trends of the food industry and offers innovative possibilities for upstream segments in the industry chain (Fig. 17).

Integrated Human Practices

Our HP is not a standalone activity separate from the project but rather a comprehensive complement to the entire project. All activities within HP are related to our project and its promotion. After discussions among team members, we have outlined the basic approach for HP: gaining a deeper understanding of the background driving project design, optimizing and improving theme design using information obtained from HP, guiding experiments through theme design, and finally assessing the effectiveness of HP through test results. Public feedback is crucial for us, so once the project design reaches a certain level of refinement, we promptly design surveys to collect societal feedback.

Meanwhile, all our HP activities strictly adhere to relevant safety guidelines.

1. Discover background and research

How Our HP Inform and Shape Our Project

A well-conducted project is inseparable from a thorough exploration and research of a solid background. Inspired by news indicating the link between high sugar intake and diabetes, we conducted on-site investigations at supermarkets to understand the application scope of sugar substitutes in reducing high sugar consumption. Simultaneously, we visited various locations, including the Nanjing Center for Disease Control and Prevention and Nanjing University of Chinese Medicine, to gather information on high sugar intake and the use of sugar substitutes. Through these investigations, we found that the current consumption of high-sugar, high-calorie foods indeed contribute to the development of various diseases such as diabetes and hypertension. The collection of this information undoubtedly provides inspiration for our project, demonstrating its value and necessity.

How We Respond to Our HP

Drawing insights from the commonly used sugar substitutes identified through the HP process and extensive consultations with professors and experts in the field, our team engaged in multiple discussions with educators to define the project's overarching direction. Ultimately, we decided to focus on producing widely accepted sugar substitutes and selecting microorganisms that align with both market demand and scientific principles.

2. Looking for Suitable Sugar Substitutes

How Our HP Inform and Shape Our Project

To effectively address the safety and economic issues of sugar substitutes and come up with specific solutions, we conducted multiple market surveys, visited several universities and companies, and consulted with experts from related companies. In each communication, we stuck to our own direction, fully considering how to enhance the positive impact of the project on society and benefit more people.

Teacher Wang pointed out that finding substitutes for sugar is one of the best solutions to the problem, and natural sugar substitutes may be a safe, effective, and promising alternative. Additionally, for diabetes patients, adding natural sugar substitutes to food can significantly reduce the impact of elevated blood sugar, undoubtedly confirming the health benefits of natural sugar substitutes for diabetes patients. Through further communication with Professor Shi from Nanjing Normal University, we learned that using Escherichia coli to produce xylitol and Yarrowia lipolytica to produce erythritol is a more suitable choice. Furthermore, the biosynthetic approach is more suitable for large-scale commercial production, offering advantages in saving time, economic costs, and increasing production.

How We Respond to Our HP

The information obtained at this stage is crucial for the design of our project. We listened to advice from various experts, selected xylitol and erythritol as the objects of our team's project, and chose E.coli and Y.lipolytica as the starting strains, conducting in-depth research. Ultimately, we finalized the specific direction of the project: using E. coli to produce xylitol and Y.lipolytica to produce erythritol.

3.Project optimization

How Our HP Inform and Shape Our Project

In order to optimize and refine the design of the project content and provide reliable guidance for experimental design, we conducted multiple interviews with expert professors in the relevant field. For example, Teacher Chen Yuan suggested the utilization of erythritol-inducible promoters for the construction of the screening system. Professor Xue Feng recommended exploring enzymes from different sources in the synthetic pathway to enhance the production of erythritol.

How We Respond to Our HP

This HP event ran concurrently with our experiments, facilitating the integration of HP with our practical work. In instances where issues arose, HP played a crucial role in problem-solving. Building upon the recommendations provided earlier, we refined the project design, resulting in a successful increase in yield of xylitol and erythritol.

4. Results of Our Questionnaire and Public outreach

How Our HP Inform and Shape Our Project

The analysis of questionnaire results and public awareness campaigns has provided us with insights into societal perceptions, attitudes toward the use of sugar substitutes, and the level of acceptance of sugar substitutes in the food industry. These analytical findings align well with the results of our earlier market research, affirming the relevance and necessity of our research.

How We Respond to Our HP

The questionnaire data and the analysis of public awareness campaigns have guided the development direction of our project design. Additionally, we leveraged these valuable social survey data in establishing a partnership with a biotech company, opening up possibilities for future collaborations.

Value orientation of the project

The value orientation of our project revolves around addressing key concerns related to sugar substitutes, with a focus on safety, cost-effectiveness, and public awareness. Here are the key aspects of the project's value orientation:

1. Safety Concerns: The primary objective is to explore and promote the use of natural sweeteners as a safer alternative to artificial substitutes. By choosing natural sweeteners, we aim to mitigate health risks associated with the consumption of artificial sweeteners, contributing to the overall well-being of consumers.

2. Economic Viability: The project aims to provide a cost-effective solution by utilizing natural sweeteners. This involves exploring ways to produce these sweeteners in a more economically efficient manner, making them accessible to a wider population. This emphasis on cost-effectiveness aligns with the goal of creating sustainable and affordable alternatives.

3. Public Awareness and Education: An essential aspect of our project is to educate the public about the benefits of natural sweeteners and dispel any misconceptions. Through surveys, outreach programs, and informational campaigns, we seek to enhance public understanding, enabling individuals to make informed choices regarding sugar substitutes.

4. Industry Impact: Considering the interests of various stakeholders, including communities, transportation hubs, and commercial establishments, the project is designed to have a positive impact on these sectors. The goal is to align with industry standards and regulations, ensuring that the adoption of natural sweeteners benefits both businesses and consumers.

5. Ethical Considerations: The project's focus on utilizing microorganisms ensures that ethical considerations related to genetic modifications and environmental impact are minimized. This approach aligns with ethical standards in biotechnology and synthetic biology, emphasizing responsible research and development practices.

In summary, the value orientation of the project is centered on safety, economic viability, public awareness, industry impact, and ethical considerations, aiming to create a positive impact on health, accessibility, and societal well-being.

Close the loop!

In our HP campaigns, a significant emphasis is placed on translating suggestions from others into the development of our themes, creating a seamless loop that aligns others' expectations with our designs. The positive feedback received is visually represented in the Figure below.