Engineered E. coli producing rhamnolipid for green enhanced oil recovery
Safety
Your Training
Have your team members received any safety training yet?Yes, we have already received safety training.
Please briefly describe the topics that you learned about (or will learn about) in your safety trainingWe learned about basic principle of biological safety including Laboratoy safety (Laboratory Acquired Infection (LAI), and Safety Regulation on Microbiological Hazard, Health and safety microbiology. It also describe about Safety Equipment, Waste Management, Recombinant DNA Technology and Biological Safety.
Please give a link to the laboratory safety training requirements of your institution (college, university, community lab, etc). Or, if you cannot give a link, briefly describe the requirements.
Our institution has a committee dealing with environmental security, health, and safety (K3L), but their main concern is not in biosafety of modified organisms. In addition, our biosafety committee (K3L) deals only in laboratory safety and environment security.
These are some example links to our university guidelines:
Each school and faculty in our institution has their own safety regulation based on different educational background, so school and faculty also has their internal man-in-charge who concerns in biosafety application like Laboratory Assistant, Research Assistant, and Technician.
But, specifically for biosafety in our institution, every year our institution has cooperate with other organization, for example, Indonesia Microbiology Association Bandung Branch to hold workshop or introduction about biosafety regulation for people who want to learn more about it, not only for college student but also for our institution employee.
Your Local Rules and Regulations
Who is responsible for biological safety at your institution? (You might have an Institutional Biosafety Committee, an Office of Environmental Health and Safety, a single Biosafety Officer, or some other arrangement.) Have you discussed your project with them? Describe any concerns they raised, and any changes you made in your project based on your discussion.Laboratory Assistant, College students and Technician.
In your country, what are the regulations that govern biosafety in research laboratories? Please give a link to these regulations, or briefly describe them if you cannot give a link.Yes, our country does. It can be accessed in http://indonesiabch.or.id/tentang-bkkhi/
Any further comments about your lab : So far our lab safety condition is quite good and can be handled well too.
About Our lab
What is your chassis organism?We use E. coli DH5alpha for cloning and E. coli BL21 (DE3) for expression.
Do you plan to experiment with any other organisms, besides your chassis? What organisms, and what experiments will you do? Please explain briefly. Please include the names of species / cell lines / strains.We planned to work with Pseudomonas stutzeri to isolate rhlA and rhlB genes by PCR. But after 16S rDNA sequencing, our cultures turned out to be Halomonas elongata and Bacillus subtilis. We later required a chromosom isolate but it turned out to be Enterobacter sp. From the three genomes, we was only able to got partial sequence of rhlA gene from Bacillus subtilis, but we don't use it make a new part.
How will your project work? Describe the goal of your project: what is your engineered organism supposed to do? Please include specific technical details and names of important parts. The RhamCOLIpid is supposed to produce rhamnolipid. Under induction, it will express rhlA & rhlB enzymes responsible for rhamnolipid biosynthesis. As rhamnolipid produced will be excreted into media, we will be able to separate it for further testing on its surfactant ability. The RhamCOLIpid will also have devices that enable user to visually observe the production of rhamnolipid in the form of colour change.
What risks does your project pose at the laboratory stage? What actions are you taking to reduce those risks? If you are working in a biology lab, you cannot answer "no risks". Even the simplest experiment, with the safest bacteria, poses some small risk. The actions you take to reduce that risk would include safety level 1 procedures, wearing rubber gloves, sterilizing waste, etc
Working with E. coli has contamination risk. The wild-type E. coli DH5 Alpha and BL21(DE3) may irritate the skin, eyes, respiratory tract, blood circulation, and kidney. To minimize the risk, same as the protect needs from ethylene glycol our team and the researchers in our lab always working safely by using gloves, lab coats, mask, and goggles (if necessary). We also apply lab safety such as sterilising the bench before and after work, sterilising waste, etc. Furthermore as our transformants will have drug resistance gene, we apply the safety so they will not escape into the environment.
How would your project be used in the real world?Our RhamCOLIpid will be used in production of rhamnolipid. Our product rhamnolipid has a wide variety of application that can use in several sectors, because a lot of sector need surfactant which can serve to reduces surface tension between two different solution, and increase solubility in water that environment friendly. Our final product will be the rhamnolipid substance, not the bacteria, so it can be released into environment more safely.
What risks might your project pose, if it were fully developed into a real product that real people could use? What future work might you do to reduce those risks?Our idea is to produce rhamnolipid in a controlled area such as in factory. The risk is that the bacteria, carrying drug resistance gene(s), will be able to escape to the environment and spread the resistance to natural microorganism. To reduce this risk, careful handling and lab safety application will reduce the risk.
