Team:Berkeley/Safety

All of our team members were given a thorough safety training prior to the start of the iGEM program. UC Berkeley EH&S office (link to eh s) closely supervises research practices on campus, and our project was first approved by this office before its commencement. **Safety forms were approved on 9/18/13 by David Lloyd and Julie McNamara.



Our team worked only with the Escherichia Coli and Saccharomyces Cerevisiae species. E. coli K-12 is not considered a human or animal pathogen nor is it toxicogenic. Any concerns for E. coli K-12 in terms of health considerations are mitigated by its poor ability to colonize the colon and establish infections (http://epa.gov/oppt/biotech/pubs/fra/fra004.htm). S.cerevisiae is a GRAS organism (Generally recognized as Safe). Saccharomyces, as a genus, present low risk to human health or the environment. Criteria used to differentiate between species are based on their ability to utilize specific carbohydrates without relevance to pathogenicity (http://www.epa.gov/biotech_rule/pubs/fra/fra002.htm). Preventive measures such as sterilization of materials prior to disposal are in place to avoid any unforeseen risks.

We do not envision any of the parts we are using to be a safety concern in the lab. All of the parts we have cloned and expressed as well as their corresponding substartes and products have been documented previously by other authors. FMO's product, indigo, is known to be a low toxicity compound (LINKhttp://www.ncbi.nlm.nih.gov/pubmed/3598883) and many of the safety practices surrounding use of indigo have been well documented in the literature.
In our envision dyeing process, indican would be the primary chemical that would coat jeans. Once indican was adhered to the jeans, our B-glucosidase would be added either as cell culture or purified enzyme to the jeans, which would then turn indican into indigo. Once the jeans have been sufficiently dyed, we have two priorities to ensure safety: removing any trace of bacteria from the cloth and removing indican. Indigo has already been proved to be safe to handle by both industry and labs. To remove both bacteria and indican, in the indican dyeing process, several rounds of washes in detergent as well as autoclaving the cloth would be ideal solutions. Because indican is soluble, it will wash off easily in detergent and water. E. Coli, the organism in which this biosynthetic dyeing pathway will be implemented, would also be killed by autoclaving because of the high temperatures (link of proof here).
We followed our these outlined safety procedures in our own dyeing of lanyards with indican. We washed our lanyards with 5% SDS (a detergent) several times and then autoclaved the cotton material before washing again with 1% detergent water.
Scaling our process would present the added safety concerns of building and maintaining a bioreactor facility. Any facility meant to grow indican-producing and B-glucosidase producing bacteria as outlined on our Human Practices page would follow standard bioreactor design regulations as well as waste and operation regulations.