Introduction

Safety has always been the most critical part of our project. Our team consistently believes that safety is a prerequisite and guarantee for the success of a project. Therefore, awareness of safety is implemented throughout the entire project. Adequate safety, security and training are provided in the use of reagents, the working environment and the life of personnel.

Project Safety

Microbiological Safety

Fig 1. Microbiological Safety(BL21), Microbiological Safety(DH5α)and Microbiological Safety(KT2440)

1. According to ATCC, the chassis organisms we have selected, such as Escherichia coli BL21 and Pseudomonas putida KT2440, can be used in a BSL 1 laboratory. The general principles of microbiological and biomedical laboratory biosafety specify that BSL 1 is appropriate for microorganisms known to have no pathogenic effects on healthy adults.

2. E.coli DH5α are used in our experimental processes. DH5α is a preferred choice for cloning vectors, Our use of the bacteria strictly adheres to experimental operating procedures and laboratory safety guidelines.

Reagents Safety

Potentially harmful reagents involved in our experiments include the inducer IPTG, Fe2+ heavy metal ions, sulfuric acid, hydrochloric acid, and organic solvents such as terephthalic acid and chloroform. IPTG is safe as a commonly used inducer. In the culture of Pseudomonas aeruginosa, in order to satisfy its need for various growth micronutrients to cope with the harsh environment, we configured micronutrient solutions with Cu2+, Fe2+,Co2+, etc. and adjusted the pH with concentrated hydrochloric acid to ensure the complex metabolic needs In addition, in the expression experiments, we used organic reagents, such as p-anisodic acid as the only carbon source, PET powder for the In addition, we used organic reagents such as dicarboxylic acid as the only carbon source, PET powder for identification of enzyme activity, sulfuric acid, and anthrone to configure anthrone sulfate solution for verification of product function. In the course of the experiments, we took adequate precautions and established records of the use of all experimental equipments and hazardous drugs to ensure that the laboratory materials would not be abused or misused.

Working Environment Safety

Our project was designed to use biological methods to degrade microplastics in water bodies and produce rhamnolipids that have plant growth promoting properties. To ensure that the bacteria do not leak into the outside environment, we use a closed fermentation environment, where the exhaust gases and inputs and outputs of the hardware are passed through a filter membrane, and UV lamps irradiate the inlet of the water streams and the outlet of the products in order to prevent any accidental release of the engineered bacteria. For the culture medium involved in the fermentation process, it is uniformly collected and strictly sterilised to prevent the escape and spread of the engineered bacteria

Lab Safety

Biosafety Laws in China

All of our project personnel strictly comply with the Biosafety Law of the People’s Republic of China implemented in 2021.

Fig 2. Biosecurity law

Training

The personnel in the project have all undergone training on relevant safety guidelines before conducting experiments. They have also participated in centralized training sessions on experimental skills and related precautions.

Fig 3. Lab safety training

Basic Lab Rules

1. Before starting work, it is essential to clearly identify the location and operating procedures of fire extinguishing equipment, electrical switches, and other facilities. Familiarity with emergency response measures and procedures is also necessary.
2. When entering the laboratory, lab coats must be worn, and avoid wearing open shoes. Lab coats should be removed when leaving the laboratory. eanliness and orderliness in the laboratory, and promptly clean up or dispose of waste materials.
3. When conducting experiments that may pose a risk, appropriate personal protective equipment must be worn as per the situation. This may include wearing safety goggles, face masks, and protective gloves.
4. Smoking, eating, and sleeping are strictly prohibited inside the laboratory.
5. Maintain cleanliness and orderliness in the laboratory, and promptly clean up or dispose of waste materials.