Our project aims to efficiently express the glucose-lowering peptide Vglycin through microbial fermentation using Pichia pastoris. The biological experiments inevitably involve safety issues, and it is crucial to ensure the safety of both the operators and the environment. We address safety in the following three aspects:

1. Bio-safety: We choose Pichia pastoris as the host to produce the peptide Vglycin, as it is a human- and environment-friendly organism.

2. Experimental safety: In the experiment, we strictly adhere to standard operating procedures (SOPs) to avoid damage to instruments, ensure operator safety, and prevent drug leakage.

3. Safety training: The lab instructor provides us with experimental safety training and offers guidance when we conduct experiments.

The materials we use are synthesized by a company and pose no risk of pathogens. We express them in organisms classified as RISK GROUP 1. Therefore, there is virtually no risk to humans or the environment. Pichia pastoris is a eukaryotic microorganism classified as risk group 1 in the IGEM white list and is not hazardous to healthy humans. We store its seed cells in a refrigerator at -80°C and immediately transfer them to an ice box when using them. We wear gloves and masks to protect ourselves during related experiments. The vector for transformation is circular DNA, which is non-toxic and safe. The plasmid is stored in a refrigerator at -20°C, and when we use it, we take it out and place it in the ice box.

Our laboratories are clean and well-equipped with autoclaves, ovens, various freezers, and more.

We wear lab coats and avoid wearing slippers in the lab. Female lab members tie their hair to prevent it from getting caught in equipment or coming into contact with hazardous materials. We regularly clean the laboratory and prepare pipette tips to maintain a good experimental environment. Before using the clean bench, we disinfect it with ultraviolet radiation and then spray 75% alcohol on gloves and necessary items.

When using the autoclave, we first add a certain amount of purified water to the sterilizer to increase the temperature and pressure of the water vapor in the pot. We ensure that the temperature of the sterilization pot drops below 70°C and the pressure decreases to 0 pascal before opening the lid. Otherwise, the internal pressure may be too high, causing steam to spurt out and potentially scalding the operators.

We conduct experiments in strict accordance with laboratory safety regulations and refer to protocols as needed (e.g., HBUT-Wuhan - iGEM 2024).

Before starting our experiments, we undergo laboratory safety training, which covers experimental operations, instruments, etc. Team members are required to pass specific exams before entering the laboratory. We also attend a 'safety education class' delivered by our instructors, where we learn the specific rules to follow in the lab and how to handle potential emergencies. During our experiments, our instructor supervises our operations, reducing the risk of unnecessary danger and damage. Additionally, our team members remain vigilant about general biosafety standards in synthetic biology and regularly communicate with our instructors to discuss and optimize product safety.

Our work is conducted in strict accordance with relevant laws, regulations, and campus safety rules. The Ministry of Education of the People's Republic of China has released the "Safety Regulations for Higher Education Laboratories" and distributed it to all higher education institutions.