Difference between revisions of "Team:CCU Taiwan/Description"

We are aiming to create a new material that will be used in paper cup coating, which is eco -friendly and similar to lignin, we call this kind of film “LIGGREEN “. We hope that cups coating with LIGGREEN are waterproof and heat-resistant; furthermore, they can be naturally decomposed. It is a breakthrough attempt for the current paper cup coating and hopes to replace petrochemical materials such as Polypropylene and Polyethylene.

LIGGREEN, which is similar to natural lignin found in most of the plants, is a polymer mainly composed of monolignol like lignin. Monolignols can go through polymerization by adding two enzymes—peroxidase and laccase. Therefore, we aim to modify the yeast to produce these two enzymes. These enzymes react with monolignols and synthesize LIGGREEN

Monolignols we chose, which are extracted from black liquor and react with our enzymes to form LIGGREEN. Then, we observe lignin’s properties and adjust it to fit lamination paper industry. With LIGGREEN, everyone will be able to enjoy the convenience without polluting the Mother Earth.

There are four parts below which are the most important for our project:

1. P.Pastoris transform

   In lignin synthesis, monomers react mainly with two enzymes, peroxidase and laccase. Then, we decided to synthesize these enzymes through synthetic biology. But we have a lot of problems from production. These are mainly about the systematic differences between Prokaryotes and Eukaryotes. Finally, we choose P.pastoris as our strain. Because P. pastoris is a high expression engineering strain, many studies have shown that P. pastoris can secrete enzymes of plant. Then the most important point is that it can modify the N-link glycosylation of our enzymes.

2. Enzyme production

   We mainly produce three enzymes, Lac1, Px16 and Px18. In gene design, we added three different tags and α-factor to help our production. Due to three different tags, we can accurately distinguish three enzymes in western blot. Then, adding the exocrine gene, α-factor, is for making a more faster production . In order to increase the value of the production line, our enzymes are not removed by cell rupture. It is also more convenient for protein purification.

3. Monolignol polymerization

   In the polymerization, the monomer mainly bonds by three enzymes. At present, we are based on the bonding of monolignol G. Between the monolignol and these enzymes, there are three types of bonds , β-β, β-O-4 and β-5. In plants, monolignol G are also polymerized with these bonds. Through the biological specificity of the enzyme, we speculate that our products have a certain relationship with lignin.

4. Business model

   Besides analyzing survey we had spread, we also consulted enterprises and professors about building business model, including key partners , channels, cost and revenues, etc., which would make our product be more completable for our customers.