Human Practices | SDU-CHINA

Introduction

In today's world, people are constantly enjoying the conveniences that plastic brings. But with it comes a whole host of problems. The threat of plastic pollution and microplastics is akin to smoke in our lives—seemingly ethereal but undeniably real.

Faced with the problem that plastics are difficult to degrade in our daily lives, we investigated the situation of microplastics in the soil, and at the same time the corresponding questionnaire survey was conducted which could help us understand the current people's understanding of degradable plastics and the dangers of plastics. To solve this problem, we have two options: degrade the existing plastic buildup or replace it with biodegradable plastic. After weighing the two options, we have chosen a more fundamental solution to address the problem: improving the production of degradable plastics.

Among a variety of degradable plastics, PHB has become our choice because of its high biocompatibility and degradability. However, it still faces many problems in the production process, such as high cost and pollution caused by crushing bacteria after production. In order to solve these problems, we have designed our three-layerdynamic regulation model to regulate PHB production.

At the same time, we are working towards the industrialisation and commercialisation of PHB products. Throughout this Integrated Human Practice process, we engage with researchers, stakeholders and the public to gather their feedback on our project design.

Identifying Problem

Environment

Plastic pollution surrounds us

The northern foot of Mountain Lao, the shore of the Yellow Sea and the shore of Aoshan Bay are our beautiful campus.

Fig.1 | Our beautiful Campus!

A paradise away from the city center, where the sea sparkles under the dazzling sunlight, reveals not pearls or diamonds, but something less glamorous: the dreadful presence of plastic waste.

Fig.2 | Plastic waste on the beach.

In collaboration with Shandong University's 'Life Sandbar' team, we organized beach cleanup events. We have gathered a large number of volunteers for this event. These efforts not only enhanced the beauty of our environment but also provided us with a clearer understanding of the plastic waste situation around our school.

Fig.3 | SDU-CHINA & Life SandBar

Global

"Plastic pollution is one of the greatest environmental challenges of the 21st century, causing widespread damage to environmental systems and human health," said OECD Secretary-General Mathias Coleman.

Inger Andersen, Executive Director of the United Nations Environment Programme (UNEP), said: "Today marks the planet's victory over single-use plastics. This is the most important multilateral environmental agreement since the Paris Agreement, and an insurance policy for this and future generations who may live with plastic, but will not be destroyed by it."

Fig.4 | A historic moment! (Image: UNEP)

On 22 February 2022, the Organisation for Economic Co-operation and Development (OECD) published a report entitled "Global Plastic Outlook: Economic Drivers, Environmental Impacts and Policy Options". The report pointed out that, on current trends, the amount of plastic products will double by 2060 in North America, Europe and East Asia; increase three to five times in other emerging economies and developing countries; and more than six times in sub-Saharan Africa. On 21 April of the same year, at the release of China's first national think tank research report on the concept and practice of plastic pollution control in China, Zhang Deyuan, an associate researcher at the Institute of Economic System and Management of the Macroeconomic Research Institute of the National Development and Reform Commission, said, "Data show that from 1950 to 2017, the world produced about 9.2 billion tonnes of plastic, and it is expected that by 2050, the cumulative global plastic production will increase to 34 billion tonnes, and the annual plastic waste production will be about 300 million tonnes."

Fig.5 | The more money, the more rubbish(Image: Heinrich boll stifung).

The global plastics situation is grim and a global consensus has been reached to actively address the plastics crisis.

Now, plastic reduction activities are being actively carried out all over the world.

Plastic reduction activities around the world

These 6 strains are for QS- switch characterization

The European Union:

In an attempt to reduce marine pollution, the European Council has adopted new EU-wide rules on single-use plastics. Under the legislation, substitutes for certain plastic items, including cotton buds, knives, plates, straws, stirrers and balloon sticks, will be banned in all Member States. Member states will have about two years to implement the legislation into national law. The EU aims to collect 90% of all plastic bottles by 2029, including a landmark target of 77% by 2025, and will introduce design requirements for caps that attach to plastic bottles.

France:

For example, from 1 January 2023, French fast-food restaurants will have to replace single-use plastic utensils with reusable ones. This is another new rule to limit the use of plastic products in the restaurant sector in France, following a ban on the use of plastic packaging boxes and the provision of plastic straws.

Australia:

And the Australian state of Victoria will ban single-use plastic products, including cutlery, straws and other items, from February 1, 2023. Companies that violate the rules could be fined up to $54,000 (366,000 yuan).

China:

China aims to establish a comprehensive management system for plastic products by 2025. This system will oversee production, circulation, consumption, and recycling. In line with this goal, Beijing aims to phase out non-biodegradable single-use plastic shopping bags by the end of 2024.

Global Degradable Plastics Related Policies

Nations	Dates	Element
Irish	2002	Levy of VAT on plastics.
Italy	2006	Legislation to heavily tax producers of common plastics.
China	2007	The General Office of the State Council on the restriction of the production, sale and use of plastic shopping bags notice: from June 1, 2008 onwards, nationwide ban on the production, sale, use of plastic shopping bags with a thickness of less than 0.025 millimeters; and all supermarkets, shopping malls and marketplaces to implement the system of plastic shopping bags paid for the use of plastic shopping bags shall not be free of charge to provide plastic shopping bags.
Italy	2011	Legislate a total ban on the use of non-biodegradable single-use plastic bags.
Germany Netherlands Belgium	2011-2014	Formulate favorable tax policies to encourage the application of biodegradable plastics and impose a disposal fee on traditional plastics.
China	2015	Jilin Province prohibits the production, sale and provision of disposable non-degradable plastic shopping bags and plastic tableware.
French	2017	A total ban on plastic bags in supermarkets and a complete ban on single-use plastic bags and plastic tableware by 2020.
United States of America	2017	California referendum passes Proposition 67, banning free single-use plastic bags from large supermarkets and pharmacies.
Malaysia	2017	Selangor government to fully implement Plastic Bag Free Day from January , 2017
Australia	2017	Plain plastic shopping bags have been banned in South Australia, Western Australia, Tasmania, Queensland, the Northern Territory and regional Canberra A national ban will be introduced after 2018.
Mongolia	2018	March 1, 2019 ban on the sale or use of single-use plastic bags.
Chile	2018	he "Plastic Prohibition Law" was officially enacted, and since February , 2019, all large supermarkets and shopping malls provide free or paid pl astic bags to buyers, and the maximum fine for each illegal plastic bag provided is up to 370 U.S. dollars; August 3, 2020, Chile will be a total an on plastic.
Philippine	2019	Philippine President Rodrigo Duterte has proposed a ban on single-use plastics.
Canadian	2019	Plastic straws will be banned from April 2020 and plastic bags from New Year's Day 2021.
French	2020	As of January 1, 2020, France banned the sale of some single-use plastic products, including plastic products such as disposable cotton buds, disposable cups and plates, and school cafeterias banned the use of plastic bottles for purified water.
China	2021	Promote degradable plastics according to local conditions, actively and s teadily, improve the standard system, and enhance the inspection. Testing and detection capabilities, standardized application and disposal.

Table 1 | Global policies related to degradable plastics

In such a situation, where does plastic go? Where does the future lie?

Our effort to understanding microplastic - the unseen risks

Plastic pollution is a widespread issue that has a direct effect on human health. The report's findings indicate that although plastic pollution varies across countries, no region is completely exempt. Notably, plastic fibers were detected in 94.4 percent of sampled tap water in the United States and 72.2 percent of sampled water in Europe.

Fig.6 | Microplastics around the world(Image:WWF).

At the same time, the distribution of microplastics is alarming.

Microplastics are easily transported by ocean currents and wind, and their pollution is distributed around the world.

Fig.7 | The distribution of microplastic in the world. (Peng Y et al., Proc Natl Acad Sci U S A,2022)

Microplastics are prevalent in the environment and can infiltrate the human body via drinking water, food consumption, air inhalation, direct skin contact, or other means. These microplastics also carry harmful substances that threaten human health.

Fig.8 | The distribution of microplastic in human body.

Given the challenging circumstances, our objective is to conduct a comprehensive questionnaire survey to assess the utilization and comprehension of plastics. among Chinese residents. To develop the questionnaire, we implemented a circular feedback process which is illustrated in the accompanying figure.

Fig.9 | The circular feedback process of our questionnaire.

After multiple discussions and experiments, our team drafted an initial version of the questionnaire. Our PI, Professor Quanfeng Liang, was consulted to further enhance its content. In line with his recommendation, our goal is to focus on degradable plastics to achieve a closed loop thought process. Essentially, developing degradable plastics is the cornerstone of solving the plastic problem.

Fig.10 | Professor Liangs advice

After that, we supplemented the content by brainstorming within the team.

Fig.11 | Modify our questionnaire.

In order to make our questionnaire design more professional and in line with relevant regulations, we interviewed Professor Zongfeng Sun. He gave us many suggestions on the scientific design of the questionnaire, the extensive collection of samples, and the accuracy of the questionnaire results. He also mentioned something about the protection of audience privacy, the overall structure of our questionnaire was adjusted, and the way of asking questions was more reasonable, and we finally issued the questionnaire.

Fig.12 | Professor Sun's resume

Fig.13 | The photo that we Interviewed with Professor Sun

At the same time, our questionnaire has been reviewed and approved by Taishan School of Shandong University and School of Life Science of Shandong University. Click here to view.

In terms of publishing the questionnaire, we carried out online and offline to ensure that the questionnaire audience was not limited to the group active on the Internet. We went into the open market to collect the questionnaire and also carry out the science popularization of plastic knowledge.

Fig.14 | We went into the open market.

At the same time, in order to enrich the source of our questionnaire group, deepen people's understanding of plastic pollution and enhance environmental awareness, we also carried out scientific popularization of plastic knowledge through academic exchange conferences and volunteer teaching and other rich and diverse forms.

Fig.15 | Science popularization activities.

The results of the questionnaire are as follows:

We have designed a complete questionnaire that covers everything from plastic usage to understanding microplastics, from the use of biodegradable plastics to the preference of the popularization of science. Obtained 520 questionnaires from across all age groups across our country.

We found that most people use plastic quite frequently, but have little knowledge about microplastics. Biodegradable plastics have penetrated into our lives, but the various shortcomings they face are still not recognised.

In addition, we may have found that short videos are a popular way for us to promote, which points us in the right direction

Click here to view detailed results

At the sametime, in order to better understand the distribution of microplastics, we have jointly launched a microplastics detection project with a number of schools and regions.

Fig.16 | The flow chart of microplastic detection.

In the planning stage, we discussed with AD Yuliang Huo in DUT-CHINA, Huo is the team leader of DUT-CHINA in 2021 iGEM, whose subject is about PET plastic degradation. About the way of microplastics detection, they adopted the method of making biosensors with organic pollutants on the surface of microplastics as markers. We had a discussion on the advantages and disadvantages of biological methods and physical methods. Due to the limited selection of microplastic indicators, we finally chose to use the spectral detection method for our subsequent experiments.

Fig.17: We discuss the way we detect microplastic with Mr. Huo.

Fig.17 | We discus the way we detect microplastic with Mr. Huo.

After a lot of journals reading and discussion, we worked out the first draft of the detection of microplastics project. After communicating with associate researcher Tianyuan Su, we further conformed some details about our standard.

Fig.18 | Mr. Su advice.

Fig.19 | We discuss the sampling standard with Professor Su.

After the final version was formulated, we contacted multiple teams including Hainan-CHINA, JLU-NBBMS, NAU-CHINA, OUC-CHINA, SMS-Shenzhen to jointly sample. We also detect soil from Jiangxi, Hebei, Shandong Binzhou, Shandong Jining with the help of many environmentalist.

Fig.20 | The sketch map of the sampling position.

After sampling, we conducted pre-treatment. We consulted WEIPU for sample testing methods and reached a cooperation. They conducted subsequent testing on the pre processed samples we provided and provided us with guidance on data processing.

Fig.21 | The technician from WEIPU teach us how to detect microplastic.

Fig.22 | The soil sample.

We conducted a correlation analysis between the results obtained from the questionnaire and the microplastics detection experiment, see modeling for details.

As can be seen from the microplastic detection results in the following figure, the soil microplastic abundance is higher in areas with more developed economy, large population, developed tourism, tourist density, and high environmental protection expenditure. At the same time, in areas where housing prices are too high or industrial pollution reduces the number of local residents and tourists, the amount of microplastics in the soil also decreases. At the same time, increasing urban greening rate can significantly reduce soil microplastics content.

Fig.23 | The result of a correlation analysi

Energy

Due to their durability, lightweight, and affordability, traditional plastics have become a crucial component of numerous products and packaging.

However, despite their advantages, it is important to note that plastics are derived from fossil fuels. This was highlighted in the May 2019 report "Plastics and Climate: The Hidden Cost of a Plastic Planet," published by the Center for International Environmental Law. The report suggests that plastic emits greenhouse gases throughout its life cycle from manufacturing to disposal.

As current policies promoting plastic production persist, it is likely that fossil fuel consumption will continue to rise. The World Economic Forum has reported that 4%—8% of annual worldwide oil consumption is now connected to plastics. If we continue to depend on plastics, it is projected that they will contribute to 20% of all global oil consumption by 2050.

Furthermore, it is understood that over 90% of plastic items are produced using fossil fuels, which discharges greenhouse gases during the entirety of the plastic's life cycle.

01. Mining and transportation cause problems

02. Refining and manufacturing accelerate emissions

03. Waste management is adding insult to injury

04. Plastic enters the environment and still has an impact on climate

After using plastic, individuals may intentionally or unintentionally dispose of it into the environment. Even if transported to a landfill, certain plastic items may be light enough to end up in waterways due to wind. This unattended plastic typically finds its way into the environment, where it continues to contribute to climate degradation as it breaks down.

Plastic on the surface of the ocean releases methane and other greenhouse gases, which will intensify as the plastic breaks down into microplastics. The current estimations cover only very few parts of the surface plastic, and there is no accurate accounting for the most part of plastic emissions below the surface.

Additionally, microplastics in the ocean may hinder its ability to absorb and sequester carbon dioxide. Since the dawn of the industrial age, the Earth's oceans have absorbed 20%—40% of all anthropogenic carbon emissions. Phytoplankton and zooplankton in seawater perform a vital function in the biological carbon pump by capturing carbon at the ocean surface and moving it to the deep ocean, thus preventing it from re-entering the atmosphere.

However, these plankton are being exposed to microplastics worldwide. According to laboratory experiments, this plastic pollution reduces their capacity for carbon dioxide removal from the atmosphere.

The authors contend that plastic pollution hinders the metabolic rate, reproductive success, and survival of the plankton responsible for transferring carbon to the deep ocean. Though the research is nascent, preliminary evidence suggests that plastic pollution may disturb the world's biggest natural carbon sink. Therefore, urgent and thorough action is necessary.

Inspiration

Two way to go

We aim to contribute towards changing the current state of plastic usage. Initially, we considered two ideas: degrade the existing plastic buildup or replace it with biodegradable plastic. Eventually, we decided to focus our project on degradable plastics. We sought professional guidance from Professor Quanfeng Liang. In the end, we opted to address this issue at its source by selecting degradable plastic.

PHB - A very powerful product

According to industry research and analysis by relevant institutions, Polyhydroxybutyrate (PHB) exhibits high crystallinity and has a smooth, hard, and brittle structure. Currently, companies worldwide engaged in PHB research and production include New Light Company in the United States, Tian'an Biological, Italy Bio-on, CoFCO Biochemical, Biomers, Zhuhai Maide Occurrence Technology, Beijing Micro-structure Works, and others.

Polyhydroxybutyrate exhibits excellent biocompatibility and biodegradability, making it a promising material for various applications in the medical industry, degradable plastics, disposable tableware, eyeglasses, packaging, sewage treatment, toys, and beyond. In biomedical materials, polyhydroxybutyrate is particularly useful in the development of sustained-release drug carriers and tissue engineering materials. In the realm of packaging, the degradation by products of polyhydroxybutyrate consist primarily of carbon dioxide and water. This aligns with the present focus on sustainable and eco-friendly practices and holds significant potential for future applications.

Polyhydroxybutyrate technology is more advanced in Japan, the United States, South Korea, and other countries with a wide range of application scenarios. Microbial fermentation remains the primary production method of polyhydroxybutyrate. While the material possesses good biocompatibility, it is brittle and exhibits poor thermal stability. In recent years, domestic and foreign enterprises have sought to broaden the scope of polyhydroxybutyrate (PHB) materials' applicability by improving them with additives like poly(styrene-co-4-hydroxybutyrate) (PSPHB) or introducing secondary monomers into their composition.

Polyhydroxyfatty acid esters (PHA), which are aliphatic copolyesters synthesized by microorganisms through the fermentation of various carbon sources, exhibit a diversity of structural variations. However, the slow industrialization of PHA can be attributed to its high production cost. Due to its vast array of potential applications, PHA research and development has garnered enthusiasm worldwide. Currently, PHA has evolved into its fourth generation, including PHB, PHBV, PHBHHx, and P34HB. Technological advancements have allowed for the expansion of the PHA industry. It is projected that, during China's 14th Five-Year Plan, PHA's planned production capacity will reach roughly 450,000 tons.

Polyhydroxybutyrate (PHB) is the earliest commercially available type of PHA, according to relevant experts. PHB exhibits the qualities of good biocompatibility and biodegradability, which makes it highly promising for use in various applications, such as packaging and medical materials. However, PHB's weak thermal stability hinders its marketability. However, ongoing research into modifications is enhancing the performance of polyhydroxybutyrate, which is expected to broaden its applications in the future.

Various Applications

Packaging materials: PHB can be utilized to produce biodegradable bags, food containers, and cups that align with consumer demand for eco-friendly packaging. Research reveals that currently, only 2% of packaging bags in the market use PHB, indicating a potential for growth in PHB usage for packaging.

Fig.24 | China Biodegradable Takeaway Packaging Boxes Market Size 2019-2025(Image:iRESEARCH)

Agricultural films made of PHB, a biodegradable material, can be utilized to cover farmland, encourage crop growth, and decrease land pollution. The current worldwide usage of plastic film approximates 1.95 to 2 million tons, with 75% of the total employed in China. Every year, we cover roughly 300 million mu of crops with plastic film, while the rest of the world uses a total of approximately 30 million mu. The current market price for typical plastic film is around $2,300 per ton. Currently, the usage of agricultural plastic film is increasing by 10% annually in China. The cost of biodegradable plastic film is over 20,000 yuan per ton. If the entire one million tons of ordinary agricultural plastic film used in the market is replaced with degradable agricultural plastic film, the market value would amount to approximately 20 to 30 billion yuan.

Fig.25 | The difference between degradable and nondegradable mulch film.(Image:Hanghangcha)

Medical Supplies: Polyhydroxybutyrate (PHB) is utilized in the medical field to produce biodegradable sutures, drug delivery systems, and prostheses, which lessen the need for subsequent surgeries and decrease trauma to patients. However, relevant data are scarce, and the connection between PHB and improved outcomes is not entirely clear. PHB and PHBHHx are capable of achieving efficient control of drug release in the cell, thereby reducing the number of secondary surgeries.

Fig.26 | The medical supplies made by PHB.

Disposable tableware: PHB can be processed into biodegradable disposable tableware, reducing the use of plastic and wood commonly used in everyday products. This aligns with the international call to reduce carbon emissions and promotes sustainability.

Textiles: PHB can also be utilized in clothing, hats, and socks, offering biodegradable and biocompatible options that are gentle and friendly to the skin.

Other applications of PHB include: PHB can be utilized for the production of biodegradable plastic bags, toys, automobile components, and other commodities.

Pain Point analysis

High market price

Among the visible biodegradable plastic products on the market, PHB holds the highest price due to its high cost. This presents a challenge in terms of market positioning and strategy formulation.

Degradable plastics and traditional plastics market price comparison and application direction

Plastic type	Market price (million yuan/ton)	Application direction
PLA	2.5-2.9	Foodservice ware and packaging, fibers, medical aids, etc.
PHA	5.1-6.2	Food containers, tableware and packaging, etc.
Biodegradable Plastics PBS	3.3-4.5	Film bags, injection molded tableware, etc.
PBAT	2-3.5	Film bags, injection molded tableware, lamination, etc.
PCL	4.2-4.5	Medical assistance, 3D additive, etc.
PE	0.8-1.4	Film products, tubes, injection molded products, etc.
PS Traditional Plastics	1.08-1.25	Instruments and meters, electrical appliances, stationery, packaging foam, etc.
PP	0.8-1.02	Pipe, sheet, film, etc.
PET	0.55-8.45	Fiber, bottle containers, etc.

Table.2 | The market price of PHB is much higher than other degradable plastic.

Techno-economic studies have shown that a major cost of pure culture production is the carbon feedstock, estimated to be up to 40% of the product cost. Biosynthesis of PHB from methane to cost $8.5/kg. This was extrapolated to $15.1–18.3/kg when accounting for downstream processing costs.But considering PHB can be produced for $2.00–6.50/kg and bio-alternatives are in the order of $2–5/kg, the cost would still be relatively high.

Stakeholder

In CCIC meeting, we listened to the lecture of Mr. Yuhan Bao and benefited a lot. After the meeting, we discussed the issue of our stakeholder with him and got a lot of guidance, which was finalized as the following figure.

We reduced the production cost of PHB, the most direct impact is the production of PHB manufacturers, and at the same time by the impact of cost reduction, the use of PHB to produce products manufacturers will also be greatly affected, including agricultural land film manufacturers, medical supplies manufacturers, packaging materials manufacturers and so on.

In addition, if PHB is widely used, we can achieve our ultimate goal of reducing plastic pollution in the environment. Environmental protection is a matter of concern for all of us!

Fig.27 | Our stakeholders.

Our solution - Cell intelligence for PHB production

Problem 1: The PHB production pathway conflicts with the central metabolic pathway.

Both TCA cycle and PHB production pathways use acetyl-coA as raw material, so if only the PHB production gene circuit is simply added to the engineered bacteria, the growth of the bacteria will be greatly affected, and the final result is low PHB production.

We turn to Quorum Sensing (QS) system, a traditional regulation tool in synthetic biology, for help. Quorum sensing system can automatically sense cell density to regulate downstream genetic on/off. It is independent of metabolic pathways and do not need exogenous inducers, which make it a perfect tool for our project.

Problem 2: As an intracellular product, PHB cannot be released on its own.

PHB are a form of carbon storage by bacteria. PHB products take up most of the space inside the cell, but will not released from the cell. The method of mechanical crushing or chemical solvent extraction used in traditional industry is not only expensive, but also brings great pressure to the environment, so we hope to design a auto-lysis system with specific expression time.

Esa I/R system

The Esa I/R system is quite special from traditional QS system. The EsaI/R QS system is homologous to the LuxI/R QS system and originates the maize pathogen--Pantoea stewartii subsp. stewartia. EsaR can act as both transcriptional activator and repressor.

Fig.28 | Esa I/R system

P_esaR is a natural EsaR-repressed promoter, whereas P_esaS is anatural EsaR-activated promoter. At low cell density (low ρ), EsaR binds to its esa box to turn off P_esaS and turn on P_esaS. In the presence of AHL, EsaR can bind to AHL and release from the DNA. Thus, at high cell density(high ρ), the P_esaS is turned on and the P_esaS is turned off.

Fig.29 | Our auto-lysis system

We will use the lambda phage lysis gene cassette to design our auto-lysis system.

Three-layer dynamic regulation model

All in all, We have designed a three-layer dynamic regulation model:

Fig.30 | Three-layer dynamic regulation model

In the first and second layer, we using a QS-switch to regulate the flow of acetyl-coA. At the early stage of growth, using QS-switch turn on the TCA cycle and turn off the PHB production pathway, so that acetyl-coA flowed into the TCA cycle and the cells grew. When the cell grows to a certain extent, the TCA cycle is turned off, while the phb production pathway is turned on, and the acetyl-coA flows to the PHB production pathway for PHB production.

In the third layer, we decide to use a stationary phase promoter and a auto-lysis system. The stationary phase promoter is used to regulate the expressing time of the downstream gene. When the bacterial reach the stationary phase, the promoter will turn on and then the cell lysis.

Putting our idea into market

Technology reviews and new thinking

We have confidence in our technology but recognize the work needed to translate it into production.

Our team had contacted Henglu Technology for insight into the distance between synthetic biology technology and industrialization. Henglu Technology identified numerous barriers preventing the commercialization of the technology.

Fig.31 | Henglu give us some advices.

Fig.32 | Henglu.

We have had discussions within the team regarding Henglu's question and have come up with some answers. However, there are still some doubts. We have sought the help of Professor Liang and the answer is as follows：

Question	Answer
The proposed new concept of 'cell intelligence' is interesting, but if cells have intelligence and perceive that they produce PHB, they will self lyse. Will cells resist to prevent death?	We can integrate cleavage genes into the genome to prevent plasmid loss. Afterwards, protective measures can be taken to prevent mutations.
Will self-cracking systems reduce production?	It will have a certain impact, but through our three-layer regulatory system, the separation of growth mode and production mode, allowing the bacteria to eat first before production, will significantly increase their yield.
In what way is the product extracted after the strain cleavage?	After adding a cracking system, extraction becomes simpler. The steps of mechanical or bacterial fragmentation are saved, and the equipment and related maintenance and operation costs or corresponding reagents required for this step are saved. This greatly reduces the cost of using organic solvents for subsequent extraction. At the same time, our product extraction requires less organic reagents, thereby achieving cost reduction.
Once the strains in the fermenter are completed after a round of production, a new batch of bacteria needs to be replaced, and in this process, strains and bacteria need to be continuously cultivated. How to prevent strains from mutating?	This is a question that every bacterium needs to consider. Sequencing can be performed before each expansion of cultivation, and timely observation during the fermentation process can effectively prevent this problem.
What are the incomparable advantages of PHB compared to other degradable plastics?	PHB (Polyhydroxybutyrate) is a type of biodegradable plastic that possesses several unparalleled advantages compared to other biodegradable plastics. These advantages can be attributed to its unique properties and production process, reflecting scientific literacy.Firstly, PHB is derived from renewable resources, such as plant-based sugars, which makes it a sustainable alternative to traditional plastics derived from fossil fuels. This reduces our dependence on non-renewable resources and helps mitigate the environmental impact associated with plastic production.Secondly, PHB exhibits excellent biodegradability. It can be broken down by microorganisms in various environments, including soil, water, and compost, into harmless natural byproducts. This biodegradation process occurs relatively quickly, reducing the accumulation of plastic waste in the environment and minimizing its long-term impact.Furthermore, PHB possesses desirable mechanical properties, such as high tensile strength and flexibility, making it suitable for a wide range of applications. It can be molded into various shapes and forms, offering versatility in its use across industries.Additionally, PHB is non-toxic and does not release harmful chemicals during its degradation or incineration. This characteristic ensures that PHB does not contribute to pollution or pose risks to ecosystems and human health.Lastly, the production of PHB can be achieved through microbial fermentation, utilizing genetically engineered bacteria. This process can be optimized to achieve high yields and reduce production costs, making PHB a potentially cost-effective and scalable solution for sustainable plastic production.In conclusion, PHB stands out among other biodegradable plastics due to its renewable sourcing, excellent biodegradability, desirable mechanical properties, non-toxic nature, and potential for efficient production. These advantages make PHB a promising alternative to traditional plastics, contributing to a more sustainable and envi.
Has there been any in-depth research into the potential huge market for PHB?	Extensive research has been conducted internationally to explore the potential market for PHB (Polyhydroxybutyrate) and assess its commercial viability. Here is an overview of some research findings:1. Market Size: Research indicates that the global market for biodegradable plastics is expanding significantly and is expected to continue growing in the coming years. PHB, as a biodegradable plastic, is considered to have immense market potential.2. Application Areas: Research has found that PHB has a wide range of potential applications in various industries, including packaging, agriculture, medical, and consumer goods. Its desirable properties, such as biodegradability and mechanical strength, make it suitable for replacing traditional plastics in these sectors.3. Environmental Concerns: The research highlights the increasing environmental concerns associated with conventional plastics and the growing demand for sustainable alternatives. PHB, being derived from renewable resources and biodegradable, aligns with the global shift towards more eco-friendly materials.4. Technological Advancements: Research has also focused on the development of efficient and cost-effective production methods for PHB. Advances in microbial fermentation and genetic engineering have shown promise in improving PHB production yields and reducing production costs.5. Market Challenges: Despite its potential, research acknowledges certain challenges that need to be addressed for PHB to gain wider market acceptance. These challenges include scaling up production, ensuring consistent quality, and competitive pricing compared to conventional plastics.In conclusion, in-depth research has been conducted internationally to explore the potential market for PHB. The findings highlight the growing demand for sustainable alternatives to conventional plastics and the promising attributes of PHB in terms of its applications, environmental benefits, and technological advancements. However, further research and development efforts.

Table.3 | Our answer to Henglu.

Market Analysis

The market for PHB has experienced significant growth in recent years, driven primarily by various factors, including changes in market size and growth rate:

Increasing environmental awareness:

Global environmental awareness is on the rise, and there is a growing concern about conventional plastic pollution. As a result, the demand for biodegradable plastics has increased, and PHB is a popular choice due to its biodegradability.

Government policies and regulations aimed at limiting the use of traditional plastics and promoting biodegradable alternatives have created a favorable environment for the PHB market.

Degradable plastics-related industry policies

Time	The name of the policy	Promulgating Unit	Thrust
January 2022	Action Program on Agricultural and Rural Pollution Control (2021-2025)	Ministry of Ecology and Environment, Agricultural Agriculture Ministry of Villages, Housing and Urban-Rural Development Ministry of Construction, Ministry of Water Resources, State Rural Revitalization Bureau	Integrate and support fully biodegradable film coverage, and make the agricultural film recycling action, including "promoting the orderly substitution of fully biodegradable film", one of the main tasks in the battle against agricultural and rural pollution.
January 2022	Implementation Program for the Promotion of Green Consumption	National Development and Reform Commission, Ministry of Industry and Information Technology and seven other departments	By 2025, the market share of green and low-carbon products will be greatly increased, the green transformation of consumption in key areas will have achieved obvious results, the green consumption mode will be universally implemented, and the green, low-carbon and recycling development of the consumption system will have been initially formed; by 2030, the green and low-carbon products will become the mainstream of the market, the green and low-carbon development mode of consumption in key areas will have basically taken shape, and the green consumption institutional policy system and the institutional mechanism will be basically sound.
May 2022	The 14th Five-Year Plan for the Development of the Bioeconomy	PRC National Development and Reform Commission (NDRC)	"Steady development of bio-energy, substitution of traditional chemical raw materials by bio-based materials, and substitution of traditional chemical processes by bio-processes" will be incorporated into China's biotechnology and bio-industry development goals during the 14th Five-Year Plan period.
July 2021	"14th Five-Year Plan for Circular Economy Development"	PRC National Development and Reform Commission (NDRC)	Promote degradable plastics according to local conditions, actively and steadily, improve the standard system, enhance the inspection and testing capacity, and standardize the application and disposal.
November 2020	Measures for Reporting the Use and Recycling of Single-Use Plastic Products in the Business Sector (for Trial Implementation)	Department of Commerce	Commodity retail places to start units, e-commerce platform enterprises, takeaway enterprises should be in accordance with the relevant provisions of the state to the business sector to report the use of plastic bags and other disposable plastic products, recycling.
July 2020	Circular on Solid Promotion of Plastic Pollution Control	National Development and Reform Commission, Ministry of Ecology and Environment, etc.	Requirements to strengthen the supervision and inspection of the ban on the production and sale of plastic products as well as the supervision and management of the ban on plastic in areas such as retail catering and agricultural films.
April 2020	Law of the People's Republic of China on Prevention and Control of Environmental Pollution by Solid Wastes	National People's Congress (NPC)	Penalize, at the legal level, the use of single-use plastic products that do not comply with the national regulations prohibiting and restricting the use of single-use plastic products. Encourage the research, development, production and sale of agricultural films that are environmentally degradable and harmless.
January 2020	Opinions on Further Strengthening the Control of Plastic Pollution	National Development and Reform Commission, Ministry of Ecology and Environment, etc.	By 2022, consumption of single-use plastic products will be significantly reduced and alternative products will be promoted.
April 2018	Guiding Opinions on Supporting Hainan to Comprehensively Deepen Reform and Opening Up	State Council (PRC)	Comprehensively prohibit the production, sale and use of disposable non-biodegradable plastic bags, plastic tableware in Hainan, and accelerate the promotion of the application of green packaging in the express delivery industry.
February 2018	National Standard for Courier Packaging Series	Standardization Administration of PRC (SAC)	Requirements for courier bags should be used biodegradable plastics to reduce white pollution; and accordingly increase the biodegradation performance requirements.
November 2017	Guidance on Synergizing Green Packaging in the Express Delivery Industry	General Post Office, Development and Reform Commission, Ministry of Science and Technology, etc.	At the national level, a number of measures have been put forward to promote resource conservation in the express delivery industry as well as the prevention and control of waste pollution, proposing that "by 2020, the proportion of biodegradable green packaging materials applied will be increased to 50%.
October 2017	Normative Conditions for the Agricultural Film Industry (2017 edition)	Ministry of Industry and Information Technology (abbr)	Encourage the research and development of the production and use of biodegradable mulch.
April 2017	Specialized Plan for Scientific and Technological Innovation in the Field of Materials in the Thirteenth Five-Year Plan	Ministry of Science and Technology (MOST)	Fully biodegradable materials shortlisted.
February 2013	Guidance Catalog for Industrial Restructuring (2013 Amendment)	PRC National Development and Reform Commission (NDRC)	The development, production and application of biodegradable plastics and their series of products are listed as encouraged.
February 2012	Twelfth Five-Year Plan for the Development of the Petrochemical and Chemical Industries	Ministry of Industry and Information Technology (abbr)	The development of degradable plastics such as polylactic acid (PLA) and polybutylene succinate (PRS) is proposed.
January 2008	Circular on Restrictions on the Production, Sale and Use of Plastic Shopping Bags	State Council Office of the People's Republic of China	Since June 1, 2008 onwards, in all supermarkets, shopping malls, markets and other commodity retail places to implement the system of paid use of plastic shopping bags, are not allowed to provide free plastic shopping bags.
June 2004	Renewable Energy (Draft) Act and Solid Waste Drainage "Environment Prevention and Control (Amendment) Act	National People's Congress (NPC)	Encourage the utilization of renewable biomass energy and the promotion and application of degradable plastics.
February 1999	(Decree No. 6 (99)	State Economic and Trade Commission (SETC)	A document stipulating a total ban on the production and use of disposable foamed plastic food and beverage containers by the end of 2000.

Technological progress:

The production technology of PHB has been continuously improved, and the production cost has been gradually reduced, making it more competitive. We control the metabolic pathway of bacteria through quorum sensing system, so that bacteria can full up before working, and save the process of breaking and collecting PHB through self-lysis system, greatly reducing the cost.

Business Model

Profit model

1. Profit point analysis

At present, the profit point of our PHB production mainly consists of three parts: strain sales, product sales, technical services, of which the direct sales of PHB products is the most important component. Our technology has greatly reduced the cost of PHB cracking.Please refer to modeling for details

2. Profit object analysis

According to the characteristics of the product and market positioning, the team subdivides the target group. The profit target mainly includes:

（1）Biodegradable plastic packaging manufacturers: PHB can be used to produce biodegradable packaging bags, food packaging boxes and cups to meet consumer demand for environmentally friendly packaging. We can provide to meet their production needs to provide materials and technology, the team of technicians to help purchasers use materials, fine debugging, provide customized services.

（2）Biodegradable agricultural mulch Distributor: As a raw material for biodegradable films, Medical products manufacturer: PHBS are used in the medical field to make biodegradable sutures, drug slow release systems and prostheses, reducing secondary surgery and trauma for patients.

（3）Disposable cutlery manufacturer: PHB can be processed into biodegradable disposable cutlery, reducing the use of ordinary plastic and wood.

（4）Textile producers: PHB can be used in textile products such as clothing, hats and socks, which are biodegradable and biocompatible and skin friendly.

We can provide them with cheaper plastic masterbatches to reduce their production costs.

Fig.33 | Structure of profit objects

3. Profit leverage analysis

The profit leverage of the team can be analyzed from the following two points: first, the research and development of intangible assets such as technology patents and scientific research achievements produced by PHB; The second is the sales of products and services, that is, how our team lets more customers accept the services provided by the our team, and then brings sustainable income to the our team.

In terms of research and development of intangible assets, PHB production with three layers of regulation is the core of our business. We will accelerate the pace of technology application and continue to follow up on product production technology iterations to maintain our technological advantages. Compared with similar products on the market, the biodegradable plastic has comprehensive innovation in product process and comprehensive breakthrough in product performance, and has the advantages of green environmental protection, low cost and high biocompatibility.

In terms of product and service sales, strong technical support is not enough, but also need to provide products and services to sell to customers. In addition to selling PHB products, our team also provides consulting, product experience, after-sales and other services to create the most suitable use environment for users. By providing personalized services, our team can form a stable source of customers.

4. Profit barrier analysis

The analysis of the two profit barriers owned by our team is as follows:

(1)Technical barriers

The use of our three-layer regulation can make the bacteria grow well and reproduce, reach a new height of PHB output, and our bacteria self-cracking after production, thus eliminating the artificial rupture process and reducing the cost.

(2)Gain customer advantage

The unique customer advantage of the team is mainly manifested in the fact that the research group of the team members has a number of cooperative enterprises, the team members participate in exhibitions and talent exchange meetings, and the customer resources obtained by the school-enterprise cooperation platform. On this basis, we won the trust of enterprises and recommended our products. In Shandong, we also reached a long-term cooperative relationship with raw material supply cooperation enterprises, forming a relatively stable supply chain system, so as to prevent other competitors from robbing our customers and safeguard the interests of the team.

Marketing strategy

According to the 4P analysis method, make the following analysis of the our team's products:

Fig.34 | Product 4P analysis diagram

Based on the 4P marketing theory, the our team takes customer satisfaction as the guideline, and always practices the principle of "customer first". By comprehensively applying the means of relationship marketing, brand marketing, service marketing, technology marketing and network marketing, the our team makes consumers feel at ease to use and buy with excellent product performance, accurate data measurement, reasonable price and friendly service attitude.

Price strategy

Our team's pricing strategy is cost-plus, supplemented by competition-driven and demand-driven price changes. Our team will first calculate the production cost, which mainly comes from the carbon source required in the microbial fermentation process and the labor, site, machine operation and other activities in the processing process, so as to determine the benchmark price of the product, according to the cost accounting of the financial department, the team will determine the appropriate price.With a price lower than the market price of similar products, by virtue of superior to similar products, quickly seize the market and start the brand, according to different market positioning, the total number of products to develop a reasonable price strategy. When downstream manufacturers buy a certain number of products, they can be given a certain degree of discount.

Channel strategy

Due to the constantly changing market environment, we have to keep pace with The Times in the selection and positioning of marketing channels. On the basis of maintaining the traditional sales channels, we will build online channels and realize the channel layout combining online and offline and direct sales distribution. The main means are divided into two points:

(1) Build online channels:

With the development of e-commerce platforms, online marketing is becoming more and more important in many sales channels, because e-commerce platforms will greatly reduce the time cost of consumers in the purchase process. We publish product information through Taobao, professional websites and other platforms to attract consumers to consume. In addition, we also make appointments through the network staff door-to-door service, telephone sales services, etc., to maximize the needs of consumers to create personalized services.

(2) Build offline channels:

Due to the large output of our products, many potential customers, but the market is relatively new, the enterprise has been initially established, and the financial strength is not strong, so the offline distribution channels are mainly divided into two points: first, the our team can rapidly expand the market size with the help of agents and distributors, alleviate the situation of direct sales accumulation, and disperse corporate risks; Second, through the our team's marketing and promotion department can directly connect with downstream enterprises, through the introduction of commission enterprises, the support of school-enterprise cooperation platform and the use of various exhibitions and business talks, contact large customer customize services according to needs, and at the same time establish a special file for their initial cooperation. Convenient for future communication and analysis, deepen the industry's understanding of our team and products and obtain the greatest degree of recognition. Establish good credit through service, and expand the market through publicity among customers.

(3) Advantages of online and offline integrated marketing:

a. Rapidly improve the visibility of enterprises and products

The Internet has the characteristics of wide coverage, low cost and fast transmission speed. At present, the team promotes our products through online channels such as network we-media platform and offline channels such as personal promotion and participation in exhibitions, and has obtained a large number of loyal customers.

b. Accelerate the marketization process

The advantages of online and offline integrated marketing can help enterprises expand their living space, conducive to competition, and quickly acquire users. Through online Taobao stores, offline to build dealer sales channels, our products have signed a letter of intent with a number of enterprises in the future, we plan to smoothly realize the marketization of products and continue to expand market share, and other degradable plastics to play a good cooperation war.

c. Improve the penetration of the market and enhance user stickiness

Through online and offline resource complementary advantages, supply chain sharing, precision marketing and other measures, can improve market penetration, enhance user stickiness, improve the competitiveness of enterprises. The team obtains big data through online channels such as Wechat public account and Taobao store and analyzes the big data to carry out precision marketing of PHB material products and improve the market penetration of products; Provide high quality content to users to enhance user stickiness.

Publicity measures

The visibility of our team determines the development of our team to a certain extent, especially in the key stage of our team to enter the PHB market through the combination of direct sales and commission sales, our team very much needs to expand its visibility, in order to seek greater development opportunities, to obtain the industry's general recognition, the main promotion and publicity methods are divided into two points:

Online publicity

(1) Wechat public account publicity

Introduce enterprises and products through online platforms such as Wechat public accounts. Vigorously promote the advantages of degradable plastic PHB products, application scenarios, project progress, national policies and green environmental protection, timely update the latest trends of products, transfer the business management concept of the enterprise, enhance the overall image of the enterprise, provide users with products without worry, improve user loyalty.

At present, the team has applied for the Wechat public number "PHB production creative small factory" and continue to update the latest status of the team

Fig.35 | "Rui Sodium Technology" Wechat public account

(2) Wechat mini program promotion

Through the Wechat mini program to enhance the user experience, help users more fully and more convenient comprehensive understanding of enterprises and products, improve consumer stickiness. The emergence of small programs has further promoted the rational layout of marketing promotion of enterprises, and built a new business service system, which can effectively communicate the relationship between consumers, theme content information and various goods and services; The mother of small program is Wechat, the our team can not only use the Wechat platform to get more and more data traffic, but also can greatly reduce the cost of the our team.

(3) We media publicity and cooperation

Make use of the convenience provided by the Internet to produce promotional videos about the PHB materials. Establish cooperation with bloggers who have certain traffic in chemistry and popular science, such as Zhihu and Tiktok, and introduce the products in the videos to improve product recognition and promote the products.

(4) Network new media publicity

In the survey on propaganda related to reducing the use of non-degradable plastics, the proportion of receiving short videos and television public service announcements was the highest. The team will set up accounts on short video platforms such as Douyin and Kuaishou to carry out relevant publicity, and then display product information in a way permitted by the platform for publicity.

We will also contact TV stations and other relevant units to publicize relevant knowledge and our research results.

Fig.36 | Survey on degradable plastic.

Offline publicity

(1) Exhibition publicity

As a targeted large-scale event carried out at a specific time and place, the exhibition is a good opportunity to help our team establish a brand image in the market, explore cooperation opportunities, and open up the existing market. The team actively inquire the latest degradable plastics exhibition, expo and other information on the Internet and organize team members to participate in the promotion of products and corporate image, so as to improve visibility, expand influence, while constantly learning new technologies to improve themselves. At present, the team participate in 2023 the 16th Ningbo International Biodegradable Plastics and Application Exhibition, and 2022 Biodegradable plastics and biobased plastics Forum,the team will continue to pay attention to the latest news in the future. Actively participate in them, actively promote our products, and grasp potential customers as much as possible at the exhibition.

Fig.37 | The 12th National Symposium on Young Microbiologists

Popularize our team's main products, establish brand image, enter the market and gradually penetrate, and constantly expand market share to play an important role.

(2) School-enterprise cooperation publicity

School-enterprise cooperation to create a window for the transformation of technological achievements is a "win-win" model for resources and information sharing between schools and enterprises. With the help of the cooperation platform between the school and relevant enterprises, the team actively participates in various activities and competitions held by the enterprise, obtains the opportunity of communication, arouses the interest of the cooperative enterprises, strives for the support of the capital and factory environment of the cooperative enterprises, conducts the test and improvement of the technology and process, gets the recognition of the cooperative enterprises, and conducts the testing and production with the enterprises. Through the recommendation or introduction of the cooperative enterprises to know more other enterprises, it is easier to gain trust, and produce cooperation intention and investment intention.

(3) Other ways to promote

Join environmental protection organizations and alliances.

With the help of Wechat public account, Tiktok, Bilibili, and other new media platforms, make simple and interesting videos, contact bloggers related to environmental protection and science and technology sectors to introduce and promote our products, improve the public's understanding and recognition of degradable plastics, and publicize the advantages and application fields of PHB to target customer groups to attract more partners and customers.

(4) Event promotion

The team plans, organizes and uses people or events with news value, social impact and celebrity effect to attract the interest and attention of the media, social groups and consumers, in order to improve the visibility and reputation of the enterprise or product, establish a good brand image, and ultimately promote the sales purpose of our products and services.

Technical improvements

In light of industrialization, a significant transition from laboratory to factory is required. Our team reached out to Professor Wang Xia from Shandong University to evaluate the conversion of our shake flask experiment to a fermentor and provide insightful feedback on the hardware portion of the fermentor design.

At the same time, in our laboratory fermentation, we have to take samples for glucose testing every few hours, which not only consumes huge labour costs, but also increases the risk of contamination by stray bacteria with multiple samples, and Mr. Wang Xia needle recommended us to use sensors to solve this problem.

Fig.38 | Professor Wang taught us something about fermentation tank.

We attended a workshop at Materials College focused on the physicochemical properties of materials and analyzed the advantages of PHB as a degradable plastic. This analysis informed our understanding of our market position.

Fig.39 | The workshop about the physicochemical properties of material.

We contacted the Continent Biotech group, who presented us with their concept of factory fermentation. Analysis of our current fermentation technology compared to existing market technology can aid in developing product sales strategies. Our key focus is to master our technology, utilizing the quorum sensing switch to achieve optimal bacteria growth and the self-cracking system to reduce PHB extraction costs - our competitive advantage in promoting our products.

Fig.40 | The advice from Continent Biotech group.

Fig.40: The Continent Biotech group helps us make our tech from lab to industry.

Fig.41 | The Continent Biotech group helps us make our tech from lab to industry.

Legal aid

We interviewed Min Wang lawyer, he mainly from the intellectual property rights belonging to the aspects and production licence to communicate with us to explain, he mentioned, intellectual property rights belonging to the school or the company should be agreed clearly; in the production licence, we must pay attention to environmental protection, corresponding to the waste treatment equipment must be equipped with in place, we can carry out internal testing and third-party testing.

Fig.42 | Min Wang-Lawyer

We have reached out to the independent director of Hailianjinhui Technology Co., Ltd. and the independent director of Shandong Zhongcheng Qingtai Law Firm to request the following assistance.

01. Legal advice:

They can offer legal guidance to assist in comprehending and complying with relevant laws and regulations. Additionally, they are capable of addressing any legal hurdles faced during the company registration process and providing expert advice.

02. Company structure and organization:

Legal advisors can help us determine the most appropriate corporate structure for your business needs, such as Limited Liability Company (LLC), Company Limited by Shares (Ltd.), etc. They can also assist in drafting articles of incorporation and partnership agreements to ensure the legality and compliance of the company's internal organization and operations.

03. Registration procedures and documents:

The legal counsel can guide us through the procedures and documents for registering the company. They understand the various documents and forms required for registration and can help us prepare and submit them to ensure a smooth registration process.

04. Intellectual Property Protection:

Legal counsel can help us protect our intellectual property, such as trademarks, patents and copyrights. They can assist in the filing and registration of these intellectual property rights and provide appropriate legal protection advice to prevent others from infringing the rights and interests.

05. Contracts and Agreements:

Legal counsel can draft and review contracts and agreements with suppliers, customers, partners, etc. They can ensure that these contracts and agreements comply with legal requirements and protect our rights and interests.

06. Legal risk Management:

Legal counsel can help us identify and manage legal risks. They can assess potential legal risks and provide solutions to reduce the risk of legal disputes and litigation that we may face.

They can provide us with professional legal advice and assistance, ensure the Company's legality and compliance, reduce legal risks, and protect our rights and interests.

Fig.43 | Our legal advisor.

After the above preparation, we contacted the leading group of Zibo Talent Work of the CPC and were invited by them to visit the center of Zibo New Industrial Park. In there, we visited a lot of company which put theory into practice.

Fig.44 | We visit the Zibo New Industrial Park.

Fig.45 | The Internal visit to fermentation factory.

Connecting Downstream Industries

As a medical product

We interviewed a doctor working in the front line of the hospital and local medical device manufacturer, they made a good introduction to the current application of PHB in the medical field, especially emphasizing the advantages of PHB's high biocompatibility, reducing secondary harm to patients.

Fig.46 | We talked with doctor and local medical device manufacturer.

Fig.47 | The doctor showed the PHB products use in medical

As packaging material

We are honored to work with the members of ShaiTech-China. They used Synechococcus elongatus, a cyanobacterium that performs photosynthesis, as a chassis to make a living light bulb that can glow in the dark using sunlight as an energy source.PHB, as a biodegradable plastic, makes a great container. This led to the idea of making a light bulb that would not stress the environment at all by using a shell made of PHB as a packaging material to encase the bacteria.

Fig.48 | The communication with SDU-CHIAN and ShanghaiTech-China.

As an agricultural mulch

We interviewed Professor Wei Zhang, who is at the forefront of scientific research. His research requires long-term use of mulch. He introduced the current use of mulch and pointed out that if the cost of the PHB mulch we produce is controllable, he is willing to be our regular customer.

Fig.49 | Botany Professor recognizes the PHB mulch film.

After these investigations, we deeply realize that PHB has a profound and extensive application background, and our technology has profound significance and broad market prospects.

YOU MAY WANT TO KNOW……

Since our program contains a lot of content, we purposely set up a focused synopsis to include the parts you might want to look for.

HP with EXPERIMENT

In order to make our ideas not only applicable to the laboratory, but also real projects that can be put into production, while our experiments were being carried out, we went to Henglu Science and Technology Co., Ltd, a real company that puts synthetic biology into industrial production, where we found problems that we hadn't envisioned in our experiments and the subsequent industrialization, and based on this, we had a deep reflection within the team and then sought help from experts in related fields. Here, we found problems that we had not envisioned in our experiments and subsequent industrialization, and based on this, we conducted deep thinking within the team, and then sought help from experts in the field, and finally adapted our experiments. See Technology reviews and new thinking for more details.

At the late stage of fermentation, we encountered a lot of problems during the fermentation process in the laboratory, and we had to take samples for measurement every few hours, which not only consumed a lot of manpower but also posed the risk of bacterial contamination. We approached Prof. Xia Wang, who gave us some suggestions for our fermentation process. For details, please refer to the Technical improvements.

HP with HARDWARE

In addition to the fermentation problems in the experiment, Prof. Xia Wang, for the selection and improvement of our fermenter also made a proposal for the characteristics of our bacterium, to find a responsive sensor to indicate the growth pattern of the bacteria, in the true sense of the realization of controllable CI. For more details, please refer to the Hardware.

Additionally, to better understand industrialized fermentation, we contacted Continent Biotech Group, who provided us with information on fermentation considerations from small to large volumes, as well as fermentation principles. For more information, please see Technical improvements.

HP with MODEL

In the background research section, we conducted field data testing and questionnaires to model the relationship between microplastics and microplastics, see Our effort to understand microplastic - the unseen risks.

Collision of ideas

At the beginning, we shared our projects with OUC-CHINA, so that we could understand each other better and facilitate the subsequent in-depth cooperation.

Fig.50: Botany Professor recognizes the PHB mulch film.

While doing the Education part of the reflection, we participated in the EDU Collaborative Exchange Conference organized by BIT-China.

For the general idea of HP, we have communicated with HainanU-China, OUC-CHINA, Nanjing-BioX, and JLU-NBBMS.

Fig.56: Botany Professor recognizes the PHB mulch film.

In response to the problem that our stakeholder is not clear enough, we consulted Mr. Yuhan Bao, who suggested to us that we should not be limited to the single dimension of the production of PHB companies, and that we can find our real stakeholder from the perspective of upstream and downstream of our industrialization and environmental protection.

We also participated in the CCIC conference to discuss and share iGEM experiences and issues with teams across the country.

References