| NDSU - iGEM 2022

Dye Safety

In the modern world, made possible by large-scale industrial production, methods to reduce our dependence on fossil fuels concerns their use as an energy source as well as the development of safe and sustainable materials necessary in our daily lives. With this in mind, our project, Dyenamix: A Better Alternative to Azo Dyes, aims to develop innovative biodegradable, protein-based dyes for use in clothing and fabric production. This project relies on the use of antibiotic resistant DH5α Eschericia coli to produce our synthetic protein. Containment of the resulting E. coli is essential as to not introduce antibiotic resistant bacteria outside the lab. We will further explain the details on how our project is safe for the environment and has been planned according to our institution’s regulations.

Safe Project Design

Our practices were done to ensure a safe handling of the modified E. coli in our laboratory and to guarantee strict adherence to iGEM’s “do not release policy”. In the future,our aim is for this project to be utilized in the industrial-scale production of textiles.. We have planned for this practical use with existing biomanufacturing processes. After expressing our protein based dye in E. coli, the cells are lysed and the protein is purified through immobilized metal affinity chromatography (IMAC). The lysing of cells after expression and IMAC purification will result in the recovery of only the pigment, preventing accidental release into the environment.

Laboratory Safety

Despite the low health risk of our project, numerous practices are required by our institution in order to maintain a safe laboratory environment:

Proper attire is required in the laboratory at all times which includes full-length pants and closed-toed shoes
Personal belongings are kept outside the working laboratory
Hands must be washed prior to leaving the laboratory
Contaminated waste is either autoclaved or disposed of in the burn bin
Surfaces are decontaminated upon completion of a given experiment
Laminar flow hoods are used to avoid contamination
Knowledge of where first aid kits, eye wash stations, and showers are located in the laboratory is required by all authorized personnel

Each team member participating in lab work also needs to meet these requirements:

Complete CITI biological safety training provided through NDSU
Be listed as a contributing member on NDSU’s project safety form
Follow all laboratory practices outlined above

Microorganism Safety

As mentioned previously, our project relies heavily on recombinantly producing our synthetic protein from DH5α E. coli. This strain is highly versatile in use of both plasmid amplification and protein expression and is low risk to the environment. Microorganisms are classified in biosafety levels (BSL) according to the health risk they can present to both humans and the environment.

BSL-1 organisms are not known to cause disease in humans and they present minimal potential hazard to the environment
BSL-2 organisms are associated with human disease and pose moderate hazards to personnel and the environment
BSL-3 organisms may cause serious or potentially lethal disease by exposure through inhalation
BSL-4 organisms present a high risk of aerosol transmitted infections and life-threatening disease.

DH5α E. coli is classified as a BSL-1 organism by the American Type Culture Collection (ATCC) and therefore requires BSL-1 certified laboratories for handling. It also belongs to the white list set up by iGEM for use in the competition.