Our project aims to produce salidroside using engineered Escherichia coli, thereby addressing the low extraction efficiency, high cost, and unstable yield associated with natural sources. The resulting product will serve factories and companies engaged in the production of health-supplement and cosmetic ingredients. Against this backdrop, ensuring the safety of our project has been prioritized as the foremost concern. Throughout the project, our team collaborates to identify and manage all potential risks while minimizing any hazards that could affect team members. Under the guidance of our advisor Wang Linlin,Guan Jiayue and Alex this document outlines the safety measures we have implemented across every aspect of our work.

This project exclusively employs the Escherichia coli strains DH5α, BW25113, and pUC19. No additional organisms are introduced, and no component—used individually or within the project—poses a hazard. Our engineered organisms or parts are unable to spread in the environment: after fermentation, the broth is centrifuged to collect the supernatant, and the cell pellet is subjected to high-temperature, high-pressure sterilization. All culture media used in the project are likewise autoclaved after use, and the containers are further disinfected with 84 disinfectant solution.
The laboratory employs GelRed nucleic-acid stain, ONPG (2-nitrophenyl-β-D-galactopyranoside), and ethanol (a highly flammable chemical), all of which may present safety or environmental concerns. GelRed is used for electrophoresis; although its documentation reports low toxicity in animal tests, it is still regarded as potentially harmful to human health. ONPG serves as a chromogenic substrate for salidroside assays at a low concentration (4 mg mL⁻¹), resulting in minimal toxicity risk. Ethanol is required for the alcohol burner that supports sterile work in the laminar-flow hood.GelRed is restricted to a designated contamination zone, and users must wear an additional pair of plastic gloves over nitrile gloves.ONPG and ethanol are handled with caution under direct supervision.
We do not intend to perform any forbidden activities or procedures requiring prior approval; our sole objective is to use E. coli as a cellular factory for salidroside production. We are interested in the heterologous expression of KivD (WP_038600683), par (BAG13450.2), UGT85A1 (NC_003070.9), UGT72B14 (AQD20582.1), and RrUGT33 (AUI41147.1). The corresponding sequences have been identified in the literature and synthesized by commercial vendors.

Our facility is classified as Biosafety Level 1 (BSL-1), so all materials pose minimal or no risk to human health. The lab is strictly divided into three zones: work, rest, and laboratory. The laboratory zone is further subdivided into a contamination area and a clean area. Agarose-gel electrophoresis is performed only in the contamination area with dedicated pipettes and Erlenmeyer flasks reserved exclusively for that space. All microbial-containing liquids are first collected in designated containers, treated with 84 disinfectant (sodium hypochlorite bleach), and then discharged. Before disposal, petri dishes and any other contaminated materials are also autoclaved to ensure that no viable microorganisms leave the laboratory.

To protect ourselves, we don gloves, lab coats, and secure long hair before entering the experimental area. Every team member must complete comprehensive training on the correct and safe use of laboratory facilities and pass a laboratory safety examination before being allowed to conduct any independent work. In addition, we adhere to a strict set of rules: no eating or drinking in the lab, no mobile-phone use inside the laboratory, no removal of laboratory waste to inappropriate bins, and so forth. A surveillance system above the laboratory area ensures continuous oversight.

1. Incident reporting: all laboratory accidents are logged in a centralized system.
2. Personal protective equipment (PPE): mandatory use of lab coats, gloves, and safety goggles.
3. Inventory control: physical materials are tracked, including who possesses them and their exact location.
4. Physical access control: only authorized personnel can enter the laboratory or storage areas.
5. Data-access control: restricted access to computers and databases.– Lone-worker/out-of-hours policy: procedures for working alone or outside normal support hours (currently not applicable).
6. Medical surveillance: monitoring for illness potentially related to organisms or chemicals used.
7. Waste-management system: complete decontamination of all biological and chemical waste before it leaves the facility.

We are gathering human data, including opinions, quotations, medical history, gender, behaviors, attitudes, and concerns. This research has received formal institutional approval or will obtain it before any data are collected. We have verified that no additional formal approval is required under applicable laws, regulations, or institutional policies.
We welcome visitors to tour the laboratory on the strict condition that they leave with nothing. Upon entry, visitors must don lab coats, gloves, and masks. Gloves and masks are sprayed with concentrated ethanol before disposal in designated bins; lab coats are reusable but are autoclaved first. For this year’s education and entrepreneurship efforts, we rely primarily on videos and photographs to showcase our project, ensuring that no potentially contaminated material is removed.

Our team rigorously safeguards experimental safety across biological, laboratory, and human-practice dimensions. We stringently ensure the safe deployment of our engineered bacteria in real-world settings. Moreover, our Human Practices outreach activities promote biosafety awareness among diverse age groups. Tailored measures protect every team member, and the combined safety protocols secure the successful execution of the project.