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Revision as of 12:32, 14 October 2018
Description
We aim to create a new material for laminating paper cups. This new film, which we call “Liggreen”, is similar to lignin and eco-friendly. We hope that cups laminated with Liggreen will be not only waterproof and heat-resistant but also biodegradable. It is a breakthrough attempt to replace current paper cup lamination based on petrochemical materials such as Polypropylene and Polyethylene.
Yeast transformation
In lignin synthesis, monolignols react mainly with two enzymes, peroxidase and laccase. We decided to synthesize these enzymes through synthetic biology. However, we found many problems with the enzymes’ characteristics. These are mainly due to systematic differences between Prokaryotes and Eukaryotes. Based on this, we chose P. pastoris as our strain for transformation. P. pastoris has high gene expression. Many studies have shown that P. pastoris can secrete plant enzymes. The most important point is that it can modify the N-link glycosylation of our enzymes.
Enzyme production
We produce three enzymes: Lac1, Px16 and Px18. In gene design, we added three different tags and α-factor to help our production. Due to the three different tags, we can accurately distinguish the three enzymes by western blot. Adding the exocrine gene, α-factor, allows for faster production. In order to increase the value of the production line, our enzymes are not removed by cell rupture, but by excretion. This also makes protein purification more convenient.
Monolignol polymerization
During polymerization, the monolignols bond with the assistance of the three enzymes. At present, we are working on the bonding of Coniferyl alcohol. Between the monolignols the enzymes cause three types of bindings: β-β, β-O-4 and β-5. In plants, Coniferyl alcohol also polymerizes with the same type of bonds.
Human practice
We consulted enterprises and professors about building a business model, including key partners, channels, cost and revenues, etc., to make our product more acceptable for our customers.