Biosafety involves precautionary measures to protect humans, animals, plants, and the environment from biological hazards, with a key focus on preventing the unintentional release of genetically modified organisms (GMOs). In our project, preventing gene leakage is a primary concern, especially since our final product is an enzyme preparation rather than live engineered bacteria. As a result, there will be no release of living engineered microorganisms at any stage of application. To mitigate the risk of gene leakage during the research and development process, we strictly follow biosafety protocols. Before each experiment, we conduct comprehensive risk assessments to ensure the containment of GMOs. Our procedures include using biosafety cabinets, controlled access, and sterilizing all materials to eliminate any chance of accidental release. Additionally, biological waste, particularly GMO-containing materials, is decontaminated following international guidelines. These measures align with iGEM’s biosafety standards and ensure the protection of both the environment and public health.

Our team selected Escherichia coli BL21 as the chassis microorganism for our project. This choice was based on the widespread use of E. coli as a common Gram-negative bacterium, which is naturally found in the environment and is also a normal inhabitant of the intestines of humans and warm-blooded animals. Most E. coli strains are harmless, and due to the extensive research on its growth conditions and metabolic pathways, we have a deep understanding of its genomic structure. This knowledge ensures the safety of our experiments and minimizes potential risks to the environment and human health.

We are using the BL21 strain, which is commonly used for protein expression. BL21 itself is not pathogenic and is widely used in laboratories, generally posing no threat to human health or the environment. Its lack of significant toxicity is one of the key reasons we chose it as our chassis microorganism. However, like all E. coli strains, BL21 could exhibit toxicity if it were to acquire certain virulence genes through plasmids or other means of genetic transfer. Therefore, our team follows strict biosafety protocols during its use to ensure the safety and reliability of the project. Based on existing research and prior usage, we carefully control experimental conditions to maintain the safety of both the laboratory environment and the broader ecosystem.

Laccase

For our group project, we chose the laccase gene Bpul from Bacillus pumilus because the laccase encoded by this gene has a wide range of potential applications in industrial and environmental treatment. B. pumilus is a Gram-positive bacterium widely distributed in soil, plants and water, and has been studied for a variety of industrial applications, including enzyme production. The laccase encoded by the Bpul gene does not pose a direct threat to the human body under normal conditions of use. As an enzyme, laccase is widely used in industrial and environmental treatment fields and is generally considered safe. However, as an exogenous protein, laccase may cause mild allergic reactions or irritation to the skin and eyes in high concentrations or improper handling. Therefore, during experimental operations and use, our group always strictly follows standard laboratory safety procedures and wears appropriate personal protective equipment such as gloves and goggles to avoid unnecessary contact and ensure the safety of the experimenters. Laccase has a biodegradable function in the environment and can help decompose organic pollutants such as phenolic compounds and dyes. Therefore, the laccase encoded by the Bpul gene is considered beneficial in environmental applications and can effectively reduce the accumulation of pollutants. More importantly, laccase is not environmentally persistent. It will be degraded under natural conditions and will not accumulate in the environment or cause long-term impacts on the ecosystem. Laccase mainly acts on organic molecules, so it does not directly threaten plants, animals or microorganisms. At the same time, at all stages of the project, we strictly control the scope of laccase use and ensure that every step in its application process meets safety standards. In this way, we not only ensure the safety of the experiment, but also ensure that the use of laccase is safe and beneficial to the environment and ecosystem.

Catalase

The second enzyme used in our group project is catalase, using the katA gene, which is widely present in organisms and has a very high level of environmental and biological safety. The catalase encoded by katA has the same function as the catalase naturally present in the human body and is non-toxic to humans. Catalase is widely present in all known animal tissues, especially in high concentrations in the liver, and is also present in plants and fungi. Therefore, the catalase encoded by katA is safe in our experiments and applications and does not pose a direct threat to humans. In the laboratory or industrial environment, our group strictly follows safe operating procedures, especially when handling high concentrations of catalase or its substrate hydrogen peroxide, ensuring that appropriate protective measures are taken to avoid possible irritation reactions or potential allergic reactions. Catalase is environmentally friendly. It converts hydrogen peroxide into water and oxygen, both of which are also harmless and non-polluting to the environment. Therefore, through safety-based management and strict adherence to operating procedures, we ensure that catalase is safe for humans, the environment and the ecosystem.

Cellulase

In our group project, we chose the cellulase gene Bgls from Bacillus subtilis. The cellulase encoded by this gene can efficiently decompose cellulose and is widely used in fields such as biofuel production, waste treatment and agriculture. As an exogenous protein, the cellulase encoded by the Bgls gene poses no direct threat to the human body under normal conditions of use. At the same time, cellulase has been widely used in food processing, feed additives and detergents, and is widely regarded as safe in these fields. However, like other enzymes, cellulase may cause mild allergic reactions or irritation to the skin and eyes in high concentrations or improper contact. Therefore, in laboratory and industrial environments, our group always strictly follows safety operating procedures and wears protective equipment such as gloves and goggles to ensure the safety of experimenters. In terms of the environment, cellulase is harmless to the environment. It can promote the decomposition of cellulose, accelerate the natural cycle of organic matter, and contribute to the degradation of organic matter and nutrient cycle in nature. Our group chose to use the cellulase encoded by the Bgls gene not only because it does not accumulate in the environment or cause long-term impacts on the ecosystem, but also because it can effectively improve soil quality and promote plant growth in waste treatment and agricultural applications, which has a positive effect on the environment. Therefore, it does not pose a threat to the environment. For other organisms, the cellulase encoded by the Bgls gene mainly acts on cellulose in plant cell walls, breaking it down into simpler sugars, and does not cause damage to animal or human tissues. In short, our group ensures that the application of cellulase is safe and effective through management, training and compliance with operating procedures in the project, and does not pose a threat to humans, the environment and the ecosystem.

Lipase

The last enzyme we used in our group project was lipase, which was encoded by the lipA gene from Pseudomonas sp. 7323. This enzyme has a wide range of applications in food processing, medicine, cosmetics, and biofuel production, and it poses no direct threat to humans, the environment, or other organisms under normal conditions of use. The main function of lipase is to catalyze the decomposition of lipids into glycerol and fatty acids, and this process is harmless to the human body. Lipases have been widely used in food processing and other industrial fields and are generally considered safe. However, like all enzymes, lipases may cause mild irritation to the skin or eyes in high concentrations or improper contact, or in some cases, trigger allergic reactions. Therefore, when handling lipase in the laboratory or industrial environment, our group always strictly follows safe operating procedures and wears personal protective equipment such as gloves and goggles to ensure the safety of the experimenters. The lipase encoded by the lipA gene is harmless to the environment. Lipases can catalyze the natural decomposition of lipids, help decompose oil and fat pollutants in the environment, have a positive effect on the environment, and therefore will not cause long-term impacts on the ecosystem. For other organisms, the lipase encoded by the lipA gene mainly acts on lipids and does not directly harm plants, animals or other microorganisms. Lipases are widely present in nature, and many microorganisms and organisms also produce lipases themselves, so it is harmless to other organisms in the ecosystem. In short, our group ensures that the application of lipase is safe and effective through rigorous operation and constant compliance with regulations, and does not pose a threat to humans, the environment or the ecosystem.

Our gene editing and genetic modification procedures are relatively safe, with minimal risk of gene mutation spread or genetic contamination. However, improper gene editing or genetic modification could pose safety risks, such as the unintended spread of gene mutations or genetic contamination. These risks could lead to the accidental release of genetically modified materials into the natural environment, potentially disrupting the balance of ecosystems.

To prevent these potential safety hazards, our team has implemented several gene safety measures. First, we strictly adhere to laboratory protocols. In the lab, we follow biosafety operational procedures to ensure that every step of gene editing and genetic modification is conducted in a controlled environment. All strains and genetic materials used are closely monitored to prevent accidental release. Additionally, we employ biosafety containment measures for genetically modified organisms, ensuring that these organisms cannot survive or reproduce outside the lab.

Furthermore, we have implemented multiple safeguards to prevent gene materials from spreading to the external environment via air, wastewater, or other means. At each stage of the project, we conduct environmental risk assessments to evaluate the potential impact of genetic modification on ecosystems, ensuring that the environmental impact is minimized.

In order to ensure the safety of the laboratory, the New Jeans team has taken several measures to focus on providing a safe environment for its members to experiment. Prior to laboratory operations, we require all team members to study the laboratory safety manual to ensure that everyone understands and is aware of the importance of laboratory safety. Our laboratory safety manual covers laboratory safety regulations, equipment usage guidelines, emergency handling measures, hazardous material storage and use rules, waste disposal codes, and the identification of common laboratory warning ICONS.

This manual clearly states the safety responsibilities of each team member and requires team members to receive training and familiarize themselves with all necessary safety procedures before entering the laboratory. During the experiment, team members are required to wear appropriate protective equipment and strictly follow the safety regulations in the laboratory. There are also detailed rules for the storage and use of hazardous chemicals to ensure that they do not pose a threat to the laboratory environment or personnel safety.

Laboratory waste disposal is an important part of the manual. We have established a strict classification and disposal system to ensure that the chemical waste liquid and solid waste generated during the experiment can be properly disposed of to prevent environmental pollution or safety accidents.

By establishing these safety rules, we hope to safeguard the health of our team members and the smooth conduct of our experiments.

Throughout our project, safety has been our top priority, covering multiple aspects including biosafety, laboratory safety, and product safety. To ensure safety, we implemented several measures. First, in terms of biosafety, we strictly followed laboratory protocols to ensure that all experimental operations were conducted in a controlled environment. All microorganisms and genetic materials used in the project were closely monitored to prevent any potential leakage or dissemination. We also used biosafety cabinets and ensured the centralized treatment of biological waste to prevent contamination.

Regarding laboratory safety, all team members received thorough safety training and wore appropriate personal protective equipment, such as gloves and goggles, to avoid unnecessary contact. After each experiment, we carried out rigorous sterilization and disinfection procedures to ensure the safety and cleanliness of the experimental environment.

For product safety, we thoroughly evaluated the safety of the enzymes and other biological materials we used. Through literature review and experimental validation, we ensured that these materials did not pose any threats to humans, the environment, or other organisms under normal conditions of use. Additionally, we assessed potential safety risks and implemented effective protective measures.

Our understanding of biosafety was applied throughout every stage of the project, ensuring that every step of the process met safety standards. By adhering to strict experimental procedures and safety protocols, we successfully minimized potential safety hazards. However, upon reflection, we realized that certain areas could be further optimized. For instance, we could enhance safety management and increase protective measures when dealing with high concentrations of enzymes or genetically modified materials.

Looking forward, we plan to enforce stricter control measures on the safety of genetic editing and transgenic operations, particularly in the handling and isolation of genetically modified organisms. We will continue to conduct environmental risk assessments to ensure that our operations do not negatively impact ecosystems. In the future, we will further strengthen the team’s safety awareness, improve safety management practices, and continually update and refine our safety procedures to ensure that safety remains a core focus in all future research endeavors.