Safe Project Design
As a part of the iGEM competition we worked on an antimicrobial surface coating. It was our goal to produce substances nisin (antimicrobial peptide), rhamnolipid (glycolipid) and chitosan (biopolymer). These were produced in non-pathogenic strains of E. coli DH5α, E. coli BL21(DE3) and P. putida K22440. Therefore, we used diverse old and new BioBricks to achieve this goal. BBa_K2380043 (a Chitin deacetylase), BBa_K2380042 (a chitin deacetylase NodB) and BBa_K2380001 (a Chitin synthase NodC) were used to produce chitin. We planned to use the BioBricks BBa_K1331001, BBa_K1331004, BBa_K1331006 to produce rhamnolipid. For nisin production we worked with BioBrick BBa_K1365000. All these BioBricks are considered to be safe to humans, animals and plants.
The resulting surface coating should be usable in places often touched by hands which causes accumulation of germs like door handles or various surfaces in hospitals. To consider it an application in public places we planned to work with non-hazardous substances. Therefore we aimed to use isolated products of genetically modified organisms (GMO) instead of the living microbes. By using various purification techniques (e.g. high-pressure homogenizer) we ensured to release only the products. All GMOs are inactivated during processing of the product by appropriate technical means.
Safe Lab Work
In our laboratory we took care of the common safety and precaution rules for the lab. Adequate working clothes (lab coat, gloves, safety googles,...) were obligatory. The release of GMOs into the environment was avoided by sterilization and autoclavation.
The reaction for the chemical linkage by divinyladipate requires special protections:
- As a byproduct of the reaction a gas may be released. Consequently we worked in an exhaust hood.
- The reaction proceeds at 80 °C bearing a possible risk of burn injuries.
- The reaction is quite unspecific. Therefore, the formation of uncharacterized side products cannot be excluded. To protect ourselves from potential hazards, we treated the product with care and wore personal safety protections (mentioned above).
To ensure safe shipment of our BioBricks to the BioBrick library, we sealed the plate with sealing foil. After packaging of the vessel we labeled the package with: "DNA, non hazardous, non regulated, non infectious, for research purpose only".