Difference between revisions of "Team:Aix-Marseille"

Nowadays, chewing gum is the second urban pollutant after cigarette butts. To clean chewing gum, specific engines are used but they are heavy and expensive. The time required for its used is long and they need a large quantity of water. Besides economical aspects, chewing gums have an environmental impact. They are dangerous for the wildlife such as birds. Indeed, believing that it is bread, they eat chewing gums and choke. Regarding these alarming facts, we have decided to create a new environmentally- responsible way to sanitize our streets. Our project is to create an E.coli strain that produces enzymes , which can degrade rubber polymers. To achieve our goal, we will use a combination of three enzymes: a laccase, a lipoxygenase or latex clearing protein (LCP) and a cytochrome C. The laccase known as an oxidoreductase will oxidize the cytochrome C which will be previously light excited. Coupled with this later, the LCP will degrade synthetics polymers found in the chewing gum. We know that the cytochrome C needs to be maturated and to incorporate a heme into its structure. To optimize the function of the cytochrome C, we will express the gene cluster involved in the cytochrome C maturation and a gene to produce a heme. In order to have the best chance of success, we selected different enzymes: 4 laccases, 2 LCP and 2 cytochromes C and we will try many combinations. Moreover, we would like to fuse laccases and cytochromes C. But, we were stunned by the lack of information about linker designing. Therefore, we will try to create a tool to easily design linker to make fusion proteins. Once several combinations will be properly cloned into an E.coli strain, we will purify proteins. Then, we will perform assays to test their enzymatic activities by oxygen measurement and spectrophotometry.