Prior to commencing any experimental work, all team members completed comprehensive safety training sessions, covering fundamental laboratory safety protocols. This included recognition of standard safety symbols, proper use of personal protective equipment (PPE), general laboratory rules, and correct operation of equipment. Both online and in-person trainings were conducted to ensure that everyone is familiar with experimental procedures such as microbial culture, gel electrophoresis, and the use of instruments including centrifuges, pipettes, thermal cyclers, water baths, gel imaging systems, and shaking incubators. Safety supervisors (instructors or senior staff) are always present during laboratory sessions to enforce PPE compliance—lab coats, gloves, and safety goggles—and ensure adherence to safety policies such as prohibiting food and drinks and proper waste disposal. A lab cleanup schedule is maintained under instructor supervision to ensure a tidy and safe working environment.

All chemicals and reagents used in the experiments are handled in accordance with safety data sheets (SDS) and institutional guidelines. These reagents are properly labelled and stored under recommended conditions (e.g., light-sensitive reagents in amber bottles, corrosive liquids in dedicated cabinets). Team members are trained to avoid direct contact and inhalation and to use fume hoods when handling volatile or hazardous substances. Spill management protocols and appropriate disposal methods are strictly followed.

During the fermentation process of Pichia pastoris, we use methanol as the inducer. During the operation, we conduct it in a fume hood to minimize the inhalation risk and prevent the accumulation of steam, thereby ensuring safety. Additionally, methanol is stored in a sealed container within an explosion-proof cabinet, away from any sources of fire, to reduce the risk of fire.

All laboratory instruments are operated under strict safety protocols, with special attention given to high-risk equipment such as electrophoresis units (to prevent electrical hazards and UV exposure), autoclaves (to avoid pressure-related accidents), centrifuges (requiring balanced loading and secure closure), thermal cyclers, water baths, and shaking incubators (to prevent sample leakage or equipment failure). Regular maintenance, operator training, and clear emergency procedures are enforced to minimize risks.

The yeast strain used in this project is Pichia pastoris GS115, which is classified as a Risk Group 1 organism and is considered safe for laboratory use. It is non-pathogenic and non-toxic. All culture procedures are performed following aseptic techniques to avoid contamination. After use, cultures are inactivated by autoclaving or chemical disinfection before disposal. Any genetic modifications are conducted in compliance with institutional biosafety requirements, and the strain is only used under contained conditions.

The proposed product is an enzyme (lipase) expressed in Pichia pastoris. It is intended for industrial applications such as detergents or biofuel processing. The product is biodegradable and does not pose known environmental or health risks. Should this project lead to large-scale production in the future, a biosafety containment strategy will be implemented to prevent unintended release. The product will be thoroughly characterized for safety and stability, and all applicable regulations will be strictly followed to ensure it has no adverse impact on public health or the environment.