. Human Practices .

Will:
As we have noticed the importance of forests in alleviating the climate crisis, we interviewed a young environmentalist from Greenwild Association. Through this interview, we want to learn how environmentalists have adopted a more sensitive perspective on climate change in recent years and how they have made efforts to protect forests.
Action:

“Are there any projects in the past few years that have impressed you or inspired you, especially in forest conservation and the fight against climate change?”
“One of our members was involved in a forest restoration project in a county in Fujian Province. The forests there have been badly damaged by years of illegal logging, and flooding worsens it. Thankfully, after several years of afforestation, the trees began to stabilize the soil.”
“From your perspective, what is the link between deforestation and global climate change? Does deforestation exacerbate extreme weather events?”
“We all know that deforestation is closely related to climate change. We're saying that forests are important carbon absorbers, which can reduce GHG emissions, and slow down global warming. If you cut down trees, you will increase the accumulation of greenhouse gases, leading to extreme weather. When it comes to floods, droughts, typhoons, and so on, it is also closely related to deforestation to some extent.”
“What do you think are the biggest challenges in promoting sustainable forest management and conservation policies?”
“Benefits. For many areas, deforestation means short-term economic gains. It takes time to convince local communities of the long-term sustainability benefits, especially when the economic returns are less obvious. So, when promoting these policies, it needs environmental awareness and corresponding economic alternatives.”
Notion:
Climate change is indeed linked to forests, and many people have been working on it continuously. However, it is more important to make people gain the reward of these actions so that it can better forest resource conservation. In terms of spreading awareness of environmental conservation, these relevant organizations or communities led by the young can spread the seeds of environmental protection to more people.
Decision:
To promote forest conservation, we need to learn more about how China is dealing with climate change in forest utilization and try to turn our project into a viable economic alternative. In this process, we also need to investigate further the public's understanding of paper products, which are timber's main applications, helping carry out relevant science popularization activities.

Will:
From the interview with the Greenwild Association of XMU, we have acknowledged that deforestation is a way of earning income for many areas. However, we think forests do more than that, so we want to find out more about the global forest industry in the context of the climate crisis.
Action:
We conducted an online interview with Prof. Liang. During the interview, he introduced the concept of forest ecosystem service and explained the ecological, economic, and social functions of forests. We learned that the utilization types of forests in China can be divided into public welfare forests and commercial forests. Public welfare forests mainly protect the ecological environment and maintain biodiversity, while commercial forests pay more attention to economic benefits. However, they are not opposites; they can be complementary and symbiotic through scientific management.
With the aim of professionally evaluating the potential of our project as a promising economic alternative for forest conservation, Prof. Liang recommended the life cycle evaluation (see in LCA) method for us. This comprehensive approach will allow us to assess our project's impact on energy saving and emission reduction in the paper industry, effectively bridging the gap between forestry and the paper industry.
Notion:
From our analysis of China's policies and the discussion with Prof. Liang, we have observed a significant shift from industrial civilization to eco-civilization. This transition is a beacon of hope for forest conservation. By employing the life cycle evaluation, we can measure our project's contributions to reducing GHG emissions and protecting forest resources, thereby advancing the implementation of this inspiring concept.
Decision:
We learned more about the changes in China's forestry policies and their implications for addressing climate change. We decided to use a full life cycle evaluation to evaluate our project's contributions to the paper industry and forest conservation.

Will:
We have come to understand that climate change is not just a distant threat, but a pressing issue that is directly linked to the survival of our forests. We are eager to visit the environmental monitoring department for a comprehensive investigation into how climate change and reforestation are impacting our daily lives.
Action:

We held discussions with the Zhangzhou Municipal Ecology and Environment Bureau. The staff told us that Zhangzhou has been affected by global warming recently, with frequent extreme weather such as typhoons, torrential rain, strong convection, drought, high temperature, and freezing damage. Meteorological disasters are generally severe, affecting the city's economic operation.
It is mentioned that carbon dioxide emission is the main cause of the greenhouse effect, deriving from thermal power generation, paper making, steel, and other industrial activities that consume large amounts of energy and materials. When it comes to the environmental impact of industry, we learned that the current paper industry pollution mainly includes ammonia nitrogen, total phosphorus, black alkali liquor, organic pollutants, etc., which are troublesome for relevant manufacturers.
Notion:
After this survey, we realized that we needed to find the details of paper enterprises' wastepaper recycling and treatment processes. We also noticed that the Municipal Ecology and Environment Bureau was also making efforts to create a low-carbon community in the local area. This urges us to pay attention to the citizens and take subsequent actions.
Decision:
Through our discussions, we have gained a deeper understanding of climate change and have brought attention to the carbon emission and pollution problems in the paper industry. This has also sparked inspiration for us to conduct more extensive fieldwork on the potential for a low-carbon community. We believe that we can make significant strides towards a more sustainable future.

Will:
We hope to investigate the recycling and utilization of wastepaper further.
Action:

We went to the Low-Value Recyclable Sorting Center in Xiamen to do research. Under the guidance of Ms. Liu, an enterprise staff member at this center, we observed and experienced the sorting and recycling processes on the spot. We found that most of the paper that entered the center had been polluted, and it was difficult to reclassify precisely.
The staff also said that the low-value mixed wastepaper could only be uniformly pulped and sent to downstream enterprises for downgrade treatment, bringing minimal economic benefits.
Notion:
We realized that improving paper recycling methods and expanding recycling ways can not only relieve the burden on related enterprises but also turn trash into treasure, achieving sustainable development of the paper industry.
Decision:
We found that wastepaper faces the problem of high sorting costs but low recycling value. This made us plan relevant publicity activities to raise residents' awareness of wastepaper sorting and recycling.
Meanwhile, we also had a deeper understanding of paper recycling, which is important to the renewable resources industry and the city's waste treatment. So, we plan to conduct further research on high-quality wastepaper resources entering the market.

Will:
We have learned about the paper industry's pollution problem and the difficulties of wastepaper classification and recycling. To further investigate the production process and post-treatment details of wastepaper recycling, we went to Nine Dragons Paper Company Limited in Quanzhou City for a field investigation.
Action:

The staff introduced the process of wastepaper recycling to us, which is mainly divided into pulping and papermaking. Although it is more environmentally friendly than wood pulp paper, recycling paper is still a relatively power-consuming process. The pulping and drying process consumes more electricity, which means more coal.
We also learned that most of the waste paper had been reccled for many times. In that case, the fiber treatment steps are more complicated, and the production efficiency is reduced, which is a common problem faced by China's recycled paper industry. Moreover, because the raw materials of recycled paper are generally waste office paper, the production process also needs to include the bleaching step, which not only consumes a lot of water and chemical reagents but also releases a variety of wastes, polluting the surrounding environment and bring adverse effects on the health of workers without properly handled.
After our introduction, the staff did not quite understand the usage of engineered bacteria in pulp making. They were worried that the metabolites of the bacteria themselves might adversely affect the quality of the pulp. The sterilization equipment in the factory was generally ultraviolet light or filter, which made it difficult to meet the needs of biosafety. So, the staff suggested that we continue to improve the biosafety part.
Notion:
The cumbersome fiber processing, the excessive use of chemical reagents, and the large processing burden of wastewater are worthy of note, and these are the targets of our projects. To adapt to the industry's specific situation, we also need to optimize our biosafety.
Decision:
Communication with the company allowed us to further clarify the specific processing difficulties of the wastepaper recycling process. We are preparing a return visit to exchange ideas about our project.

Will:
After investigating the recycled paper industry, we hope to gain a more specific understanding of the upstream of the waste paper recycling chain, hence we went to the Fuxing Renewable Resources Recycling Store.
Action:
The Store receives wastepaper from residents, scavengers, and sanitation workers, which can be up to a total of 600 kilograms per day. After sorting by staff, the wastepaper is sent to a downstream sorting center for compressing and packaging and finally sold to paper mills for reprocessing.
Notion:
We observed that the amount of wastepaper in the recycling station is relatively stable and large, but this is in contradiction with the declining utilization rate of wastepaper in China (details in description).
Decision:
According to this survey, we recognized that the low recycling rate of waste paper is not due to insufficient amounts of waste paper recycled, but rather a bottleneck that hinders the utilization of waste paper. To find out the real bottleneck of wastepaper recycling, we decided to seek relevant guilds to conduct in-depth research.

Will:
The interview with the Fuxing Renewable Resources Recycling Store provided new insights, highlighting the necessity of gaining a comprehensive understanding of the entire industry. To obtain more complete information, we recognized the need to connect with a network, such as an industry association, to investigate the paper recycling process.
Action:

We had an interview with Ms. Zhang, Deputy Secretary-General of the Association. We learned that different types of wastepaper differ in the degree of pollution and the purity of fibers. For example, the low-value mixed wastepaper will cause pollution due to the printing ink, adhesives, and other chemicals. This can increase the difficulty of recycling and affect the efficiency of recovery and the quality of recycled paper.
Notion:
Verified by many investigations, we finally found the reason for the low utilization of wastepaper: the low deinking efficiency. To truly achieve efficient and eco-friendly wastepaper recycling, we need to optimize the deinking step to overcome the problems of the high cost and low quality of recycled paper.
Decision:
Through the investigation of wastepaper recycling in the market, we found that there are indeed difficulties in industrial paper recycling, and the key to solving them may be optimizing the deinking step. We also started to explore a more targeted scheme of synthetic biology.

Will:
We learned that ink contains ink binders, namely resin mixtures, helping the ink attach to the paper, which hinders our de-inking process. So, we planned to use PETase and lipase to destroy the ink binders. We want to know the feasibility of this strategy.
Action:
We visited Prof. Yang and discussed the feasibility of our project with him. He said the degradation of polymeric resin by enzymes is currently very inefficient. So, it is recommended that we destroy the paper fibers in contact with ink to achieve the purpose of de-inking because destroying a small amount of cellulose on the surface is easier.
Notion:
These suggestions successfully made us start to pay attention to cellulase. When faced with a difficult problem, we can think about the solution from a different angle.
Decision:
Considering the suggestion of Prof. Yang, we further evaluated the feasibility of PETase and finally decided to use cellulase instead of PETase.

Will:
After we realized that wastepaper recycling is facing the problem in the de-inking process, we desire to search for more effective enzymes in pulp de-inking.
Action:

During the experiments, we found that the de-inking effect of cellulase, lipase, and laccase was very limited. So we showed the specific details of chemical deinking to Prof. Chen, hopefully, to find an alternative.
The key to the pulp deinking may be related to using the oxidant, which is hydrogen peroxide. So he suggested us to use oxidase, such as monooxygenase.
Prof. Chen explained to us that monooxygenase inserts a single oxygen atom in the process of reacting with substrate molecules so that the conjugated side chains of lignin and other colored substances. Finally, these colored substances become unstable, and ultimately achieve the purpose of bleaching, which may be able to achieve the same or even better effect than hydrogen peroxide treatment. This gives us a new idea for our processing system, more details can be found on our Design page.
Notion:
With the introduction of monooxygenase, we may be able to synchronize the de-inking and bleaching processes, which will further improve the production efficiency of the company.
Decision:
Through this exchange, we have managed to focus on monooxygenase and its potential bleaching effects. After the introduction of monooxygenase, the de-inking process has been significantly improved. Details can be seen on Results.

Will:
After finding some effective candidate enzymes, we hope to communicate with Prof. Zhuang about the feasibility of our release system to make these enzymes work effectively in reality.
Action:

In the initial design, we used a bacterial autolytic system called FLSA (FhuD-Lysozyme-SsrA Mediated Autolytic System) for the effective release of intracellular products (Details in Design). Prof. Zhuang expressed concerns about our autolytic system.
The leakage of plasmids is inevitable when cracking engineered bacteria without safety measures. Moreover, if adding extra inducers to initiate autolysis is applied in an industrial environment, the cost will be dramatically increased due to the large consumption of inducers.
To our surprise, Prof. Zhuang proposed a new idea – engineer the protein secretion ability of the bacteria to achieve the goal of efficient enzyme release. Because the engineered bacteria can release the enzyme continuously, rather than cracking it all at once. This inspired us to redesign our project.
Notion:
Based on the experiments of LMT in 2021 XMU-China and 2023 XMU-China, we tried to compare the LMT with the other seven signal peptides recommended by Ms. Zhuang.
Decision:
Through communication with Prof. Zhuang, we considered more details about the actual industrial treatment scenarios. We re-examined our design from the perspective of efficiency and biosafety, carrying out corresponding optimization, which made our deinking enzyme system more feasible and reliable. Finally, we decided to use the LMT to secrete our enzymes.

Will:
In the process of our experiment, we encountered the problem of fiber loss described by the Nine Dragons Paper Company Limited, and we needed to explore new ways to improve the fiber yield.
Action:

To improve the efficiency of the industrial de-inking process, we had a interview with Prof. Sha. When we introduced the project, we analyzed the industrial pulping process together with him, including pulping, screening and purification, washing and concentration, de-inking flotation and finally bleaching. Prof. Sha paid attention to the process of flotation. He explained to us that flotation is an effective way to separate solid particles in the liquid phase environment, but in the paper recycling process, the scale of ink and fine fibers is close to each other. So, it is difficult to effectively separate by flotation method.
However, most of the ink and fibers still have differences in the hydrophilic relationship, and perhaps hydrophobic solvent extraction can be more effective in separating them.
Notion:
The reason why we could not improve the separation efficiency before may be the floating method, not just about fiber quality.
Decision:
With the help of Prof. Sha, We fully analyze the properties behind the industrial treatments to better find the key and feasible solutions to the problem. Considering the environmental friendliness and biodegradability, we use limonene for ink extraction, rather than the original flotation method by surfactants, bettering the deinking process.

Will:
We hoped to use alginate hydrogel beads wrap-engineered bacteria as our biosafety strategy. On Prof. Zhuang's recommendation, we visited Prof. Xiao to exchange information about the preparation and use of hydrogels. We hope his rich practical experience can help us better evaluate the feasibility of the alginate hydrogel beads.
Action:

We had an in-depth discussion with Prof. Xiao about the biosafety strategy of wrapping engineering bacteria by using alginate hydrogel beads, especially the side effects of alginate hydrogel beads in nutrient preservation and heavy metal ion transportation.
Dr. Xiao regrettably indicated that we could not narrow the pore size of the hydrogel to avoid the dissipation of small molecules of nutrients and inducers, because it might also impact the adsorption process of heavy metal ions. At the very least, using a hydrogel carrier is not the most economical option in the industry.
Notion:
This exchange made us rethink the necessity of hydrogel carriers.
Decision:
This conversation made us think more critically consider the use of alginate hydrogel beads. Finally, we decided to abandon this scheme, and combined with the actual situation of the Nine Dragons Paper Company Limited, we decided to use the light-triggered kill switch to ensure biosafety.

Will:
After the last communication, we further improved our design. To further adapt to the industrial process of the recycled paper process, we introduced the light-triggered kill switch, combined with the deinking enzyme and the enzyme release system based on the signal peptide LMT that we screened, hoping to better meet the needs of the Company to simplify production conditions and ensure biosafety.
Action:

We once again communicated with the Nine Dragons Paper Company Limited. about the feasibility of our deinking solution. We presented with a complete and optimized deinking process and wastewater treatment solution, including efficient deinking enzymes and heavy metal ion adsorption systems (see Design for details), and established more standardized operating procedures (see SOP for details). To enable staff to better understand the results of our projects, we evaluated our solutions in terms of laboratory deinking effectiveness, cost control, and carbon emission control (see LCA for details), which was highly valued by the staff.
It is worth pointing out that although our light-controlled kill switch can be well adapted to the current production conditions of the industry. In addition, the staff have encountered great difficulties in understanding the concepts related to synthetic biology. Subsequently, we conducted an introduction on synthetic biology, introducing the application of engineered bacteria to dyes degradation, the production of high-value-added products, and so on. Refreshing the thoughts of employees about engineered bacteria has succeeded in promoting public awareness of the powerful tools of synthetic biology.
Notion:
Further interaction withthe Nine Dragons Paper Company Limited has provided significant feedback to our project, which includes not only a more comprehensive evaluation of our project but also the public's understanding and perception of synthetic biology. We are more deeply aware that the successful promotion of an application program needs people's understanding and recognition.
Decision:
Through this exchange, we realized the importance of further promoting the project, so we decided to publish relevant scientific science articles on the public account to promote public understanding and recognition of our project and stimulate public interest in synthetic biology.

Will:
Based on the previous interview and survey results, there is a lack of public knowledge about wastepaper recycling. We hope to raise the awareness of children and teenagers to cherish and protect forest resources.
Action:
We had cooperated with the Science and Technology Museum to organize a science popularization event with the theme of protecting forests and saving paper. By issuing brochures, playing games, poster displays, and project promotion, we helped children and teenagers understand the ways of wastepaper recycling and cultivate a paper-saving lifestyle.
Notion:
We have received many positive feedbacks from participants of our event. We really felt that many of them are curious about nature and thirsty for knowledge. However, most of the participants were under preliminary school age, so we then plan to carry out events among older children.
Decision:
Through this activity, XMU-China not only demonstrated scientific research achievements and innovative thinking but also reflected the iGEMer's sense of social responsibility and public welfare spirit.

Will:
Based on previous research and activities, we decided to promote science popularization for children and teenagers to raise their awareness of paper saving and forest conservation.
Action:
We came to the Yanwu Primary School to carry out the Protect the Forest and Save Paper science popularization activities, which not only included the popularization of knowledge but also included tailored experiments. In the class, Prof. Hong, the PI of our team, introduced some basic knowledge about forests to students and explained how trees simply become our writing paper.
He also explained to the students the indispensable "partner" in synthetic biology ---- E. coli, which excites the children's interest. Through picture display and video learning, the children had a new understanding of E. coli, and the infinite possibilities given by synthetic biology also make them felt the charm of science.
To let the students better understand the nature of carbon dioxide and the concept of forest protection, the team designed a variety of fun experiments in advance to help the children better understand the scientific principles behind the interesting phenomenon through the way of education and entertainment.
Notion:
By enriching our learning methods and constantly trying in practice, we cannot be limited to textbook knowledge and truly feel the charm of science in life. We also hope to stimulate students' interest in synthetic biology through this activity, so that the flame of synthetic biology will pass on.
Decision:
This activity not only enhanced the children's awareness of treasure and protection of forest resources but also let the children develop a paper-saving lifestyle. So far, with many offline activities, we decided to publish these gains in the form of popular science articles on our public account so that more people can understand and even join our actions to protect forests and cope with the climate crisis.