We chose E. coli as our chassis organism. Three lab strains were selected: “BL21(DE3)”, “W3110”, and ”W3110 (L5T7)”. These lab strains are reported to be not pathogenic to human. We worked these strains in authorized labs (biosafety level 1 lab and higher) only.

Three new basic parts with open reading frame were created in our project, which are Rubisco, prk, and CA. Three genes are derived from Synechococcus elongatus PCC 7002 and Synechococcus elongatus PCC 7942, which belong to biosafety level 1 nonpathogenic cyanobacteria.

This is the third year since the establishment of iGEM NCKU Tainan team. We have raised the awareness of synthetic biology in our campus. NCKU President Dr. Jenny Su and Dean of College of Engineering promised to provide us with a headquarter, including a meeting room and a lab. To meet the lab standard of Taiwan CDC (Center for Disease Control), we contacted with Mrs. Liu Yee Fen, the contact person of Center for Occupational Safety and Health and Environmental Protection, which is the authorized unit in NCKU responsible for lab safety and security. With the assistance of our PI, Pror. Ng, we successfully certified our lab to BSL-1, safety level one. Mrs. Liu gave us some lab security instructions and advice. Each of our members should attend the biosafety lab training program.

In addition to the lab in iGEM headquarter, the wet lab team also performed experiments in various labs whose supervisors are Prof. I-Son Ng, Prof. I-Hsiu Huang, Prof. Han-Ching Wang, and Prof. Hashimoto Masayuki. The labs mentioned have all been certified as biosafety level one or two.

It is a must for anyone to pass the authorized training session before conducting any experiments in the lab. Since our applied design must be tested with the constructed bacteria, all of our team members will be exposed to the lab. We, therefore, decided that the whole team should be certified to do the experiments in biosafety level one lab. Team members from biotechnology have already passed the training session, and the rest of team members have passed the training online course. We believe that learning safety and security knowledge could lower the risk when we conducted experiments inside a lab.

The goal of our project is to utilize CO₂ emitted from industries by using modified E. coli , which has high potential to produce valuable byproducts. At the same time, we are aware of the safety concerns of introducing synthetic organisms into the environment. We designed a bioreactor that ensures the engineered E. coli in maintained in a closed system separated from the environment. Each inlet and outlet of the bioreactor is equipped with a filter to prevent the outflow of the engineered bacteria.

We tested the safety of our device by incubating bacteria inside. We collected the waste sample from the filter and checked for bacterial growth. Comparing with the control group that does not pass the filter, we found out that the filter can effectively isolate the bacteria inside our device.

Fig 3. Result of biosafety tests with our bioreactor

In our product design, we planned to extract protein and bioproduct from E. coli. The waste should be then collected and sterilized. We planned to combine the waste heat recovery system in the factory to sterilize the waste medium. One of the many goals of our product design; to turn waste into valuable resource, has been fulfilled by this biosafety design.