As an initial step to solve the plastic waste problem, we read the literature, learned what have already been done and met with several experts in different aspects of plastic waste: Prof. Alex Sivan, a scientist from the Ben-Gurion University who is an expert in microbial degradation of plastic waste. We consulted Prof. Sivan about our approach to a new solution for plastic degradation and learned that LC-Cutinase is an enzyme that is known for his ability to break PET bonds into two monomers: ethylene glycol and terephthalic acid. After learning about the LC-Cutinase, and making a comprehensive research about its properties we decided to focus on improving its PET degrading activity.
Next, we consulted Dr. Sarel Fleishman, a scientist from Department of Biological Chemistry, Weizmann Institute of Science, Israel, who is expert in computational protein design to improve enzyme activity. Dr. Fleishman helped us design the LC-Cutinase variants in order to improve its activity. We used his recently published algorithm- PROSS (Netzer and Fleishman, Science 2016), and designed rational mutations to make a thermodynamic stable protein. We found that the coding optimized as well as few variants improved the LC-Cutinase activity and its ability to degrade PET. At the same time, we investigated the metabolic pathway that is required for our project. We found that the Pseudomonas putida bacterium has the qualities and requirements to engineer for a biodegradation product. We contacted Prof. Victor De Lorenzo, who uses advanced molecular biology and genetic engineering of microorganisms, like P. Putida, for biomonitoring and bioremediation. Prof. De Lorenzo contributed from his extensive knowledge and sent us the pSEVA plasmids that suitable for genetic engineering of P. Putida. We also contacted Prof. Dan Tawfik and Dr. Halim Jubran from the Department of Bimolecular Sciences, Weizmann Institute of Science, who provided the P. Putida KT 2440.
In addition we met Prof. Amir Aharoni and Dr. Shimon Bershtein, two scientists from our university who are expert in protein engineering who study and improve complex biological processes. They advised us how to design the metabolic pathways in P.putida and provide important advices about genetic engineering. Finally, we discussed our plan and the project's design with our PIs concluding the valuable information to start wet lab work on our project.
Furthermore, we done a comprehensive investigation of plastic's implications in order to learn and be able to raise awareness of the plastic hazard. Throughout the project, we met experts from the industry who utilize plastic in its raw state - Coca-Cola Company, Genome compiler and many more. We have learned about plastic's convenient properties such as flexibility, rigidity and lucidity that make plastic a popular raw material in the industry.
Finally, we aimed to learn about the current solutions for plastic waste. We visited several leading industrial companies, such as 'Neot Hovav- eco industrial park', 'Aviv- Recycling Industries' and 'ADAMA Agricultural Solutions' to learn about their way of dealing with plastic waste. By visiting at Aviv recycling company, we saw and learned the process of plastic recycling – from the arrival of the plastic bottle to the plant to its reduction into raw plastic material made for the creation of other products. One of the most interesting and important things we have learned is that 99% of the recycled raw plastic material is being used for products that cannot be recycled again. Discovering that the current recycling process produces plastic waste that is not recyclable was eye opener, as it emphasizes the unmet need for biodegradation of plastic waste.
With the acquired knowledge, the magnitude of damage of plastic waste, and it became clear to us that we must find an effective and fast solution to deal with this problem.