Team:UESTC-China/Safety
Overview
UESTC-China knows how important safety is to the entire project, so we divide Safety into two parts: Biosafety and Lab Safety. In the biosafety section, E.coli and Pichia Pastoris we used in our lab both belong to Risk Group 1, so they
are not harmful to human and environment. Meanwhile we understand the inherent risks of working in a lab facility and aim to take all necessary precautions to ensure no personal or environmental harm occurs. To this end, we have implemented
Lab Safety. And our completed safety form can be found here.
Project safety
Our project aims to develop a kind of mixed enzyme preparation which can be used for deinking and design the corresponding hardware. Considering the future commercial application scenarios of our products, we must take strict safety measures
to ensure that our final products are qualified. In order not to harm users and the environment, and to make Deinker a responsible product for the world, we have considered possible safety and ethical issues.
Ethical safety
We plan to commercialize Deinker as a product, but there are some ethical issues. Since our product can be used for deinking, users may use Deinker to tamper with documents illegally, which may bring other bad effects. But we can't let
potential problems stop us from creating Deinker, as every product may face this kind of problem, and many useful things won't occur if they are given up. And the detection technology for tampering with paper documents is now so advanced
that ultimately there is no harm to people or communities，because the current detection methods are also very advanced.
Biosafety
Introduction
Our Pichia Pastoris will be engineered to produce four engineered enzymes that can be used for deinking. They are cellulase, xylanase, laccase, and lipase. We added two specific cohesin domains from native cellulosome to the cellulase
and xylanase respectively, so they can combine with two other Phicia Pastoris produced proteins named scaffoldin and anchoring to become a newly designed cellulosome system for deinking. This self-designed cellulosome system, in the
end, will be anchored in our Pichia Pastoris' cell wall through anchoring protein.
Chassis
The chassis we used are Escherichia coli Trelief 5 alpha and Pichia Pastoris GS115, both belonging to Risk Group 1, so they are not harmful to human and environment. But we have inserted resistance gene into their genome and it may cause
drug resistance by HGT.
One point should be mention that Pichia Pastoris GS115 is histidine deficient and can only grow on YPD medium or in minimal media added histidine. To avoid contamination, all organisms we used in lab would be sterilized by steam under
high pressure at the end of the experiment.
Parts
The parts we extracted were all synthesized from the company without any risk of pathogens, and we expressed them in the organisms in RISK GROUP 1. Therefore, there is virtually no risk to human or the environment.
In addition, considering that the application scenario of the product is more in the office and home, we have abandoned the most commonly used methanol-induced AOX1 promoter, and adopted GAP promoter instead, which can avoid the harm to
human and environment caused by the use of methanol in the production process.
Lab safety
This year, our team completed a Laboratory Safety Handbook, which includes the basic rules and codes of conduct that the laboratory needs to abide by, as well as some emergency handling methods and other contents.Click here for more details about Laboratory Safety Handbook.
Expected Protection Mechanism
For the organisms we use, even if they are not harmful to people and the environment, we always insist on sterilization treatment at the end of the experiment to prevent them from escaping out of the laboratory.
For our lab members, we have a first aid kit in the lab, spare basic band-aids, iodine, etc. In addition, we are equipped with an eye wash and spray device 30 meters away in the laboratory to ensure that injured members can be timely cleaned.
Disposal
To avoid the pollution of environment, we gather all medium with bacteria and sterilize them before discarding them. For some reagents with health risk, such as TEMED, perchlorate, we use them carefully with the correct position and keep
another separate container for hazardous waste.
In order to keep our laboratory safe and clean, we carry out regular cleaning, sterilization, and disinfection every day. Besides, we also check out the electrical installment and instrument to ensure safe experiment and environment. Ultra-clean
working table is treated with ultraviolet sterilization before use.
Training
Before our experiment, we carried out laboratory safety training, including experimental operation, experimental instruments, etc., and the team members needed to pass certain examination before entering the laboratory for experiment.
In addition, at the beginning of the experiment, we were divided into groups of two or three, so that we could supervise each other during the experiment to prevent wrong operation.
Experimental operation
Our experimental operation is carried out in strict accordance with the following requirements:
1. Must wear lab coat, gloves and a mask when doing the experiment.
2. When volatile or irritating reagents are needed, do the test in a fume hood.
3. All reagents need to be labeled, including its name, concentration, expiration date, the person who prepared the reagent (if the reagent is prepared by ourselves) and other information. All reagents must be returned to their original positions after use.
4. During the experiment, the experimental operation shall be carried out in strict accordance with the experimental steps, and the experimental operation shall be standardized.
5. Clean up the experimental table immediately at the end of the experiment, and carry out daily cleaning and necessary disinfection of all worktables.
2. When volatile or irritating reagents are needed, do the test in a fume hood.
3. All reagents need to be labeled, including its name, concentration, expiration date, the person who prepared the reagent (if the reagent is prepared by ourselves) and other information. All reagents must be returned to their original positions after use.
4. During the experiment, the experimental operation shall be carried out in strict accordance with the experimental steps, and the experimental operation shall be standardized.
5. Clean up the experimental table immediately at the end of the experiment, and carry out daily cleaning and necessary disinfection of all worktables.