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

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<div class="photoDescription"><h1 class="bigtitle">DESCRIPTION<h1></div>
 
<div class="photoDescription"><h1 class="bigtitle">DESCRIPTION<h1></div>
 
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             <br><p class="description">&emsp;&emsp;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.  
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             <br><p class="description">&emsp;&emsp;Our product, “Liggreen” is a new material that can laminate on paper cups to replace current paper cup laminated with petrochemical materials such as polypropylene and polyethylene. Liggreen is a lignin-like polymer. Cups laminated with Liggreen is supposed to be waterproof, heat-resistant and eco-friendly.
 
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             <p class="first"  id="ca1">Yeast transformation</p>
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             <p class="first"  id="ca1">Enzymes</p>
             <p class="description">&emsp;&emsp;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.</p>
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             <p class="description">&emsp;&emsp;n lignin synthesis, monolignols react with two enzymes, peroxidase and laccase. Liggreen, is modelled on Picea abies, which is the most common spices used in forestry and possess high content of G which is the cheapest monolignol so that it can cost down our product. We selected peroxidase gene Prx16 and Prx18 from Picea abies, laccase gene from Pinus taeda. Pinus taeda and Picea abies are both gymnosperms and have similar ratio of monolignols. Moreover, the reason why we choose laccase from Pinus taeda since it has highest reaction activity at pH 5.0 which is corresponded with peroxidase react with monolignol in vivo. </p>
 
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             <p class="first" id="ca2">Enzyme production</p>
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             <p class="first" id="ca2">Host</p>
             <p class="description">&emsp;&emsp;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. </p>
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             <p class="description">&emsp;&emsp;We produce laccase and peroxidase by modified P. pastoris (SMD1168). There are three main reasons why we select P. pastoris as our host to produce these two enzymes. First and foremost, laccase and peroxidase are types of glycoprotein that required N-link glycosylation to modify our enzymes, so eukaryotes are more fit than prokaryotes. Second, many researches have applied P.pastoris to secrete plant enzymes. Last but not least, P.pastoris can yield large quantity of protein, it is exactly beneficial for us to produce our product in a large scale. Moreover, we let P.pastoris secret the enzyme by adding a secretion peptide so that we do not need to disrupt the organism in order to decrease the consumables of the following production cost. </p>
 
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Revision as of 19:27, 16 October 2018

DESCRIPTION


  Our product, “Liggreen” is a new material that can laminate on paper cups to replace current paper cup laminated with petrochemical materials such as polypropylene and polyethylene. Liggreen is a lignin-like polymer. Cups laminated with Liggreen is supposed to be waterproof, heat-resistant and eco-friendly.

Enzymes

  n lignin synthesis, monolignols react with two enzymes, peroxidase and laccase. Liggreen, is modelled on Picea abies, which is the most common spices used in forestry and possess high content of G which is the cheapest monolignol so that it can cost down our product. We selected peroxidase gene Prx16 and Prx18 from Picea abies, laccase gene from Pinus taeda. Pinus taeda and Picea abies are both gymnosperms and have similar ratio of monolignols. Moreover, the reason why we choose laccase from Pinus taeda since it has highest reaction activity at pH 5.0 which is corresponded with peroxidase react with monolignol in vivo.

Host

  We produce laccase and peroxidase by modified P. pastoris (SMD1168). There are three main reasons why we select P. pastoris as our host to produce these two enzymes. First and foremost, laccase and peroxidase are types of glycoprotein that required N-link glycosylation to modify our enzymes, so eukaryotes are more fit than prokaryotes. Second, many researches have applied P.pastoris to secrete plant enzymes. Last but not least, P.pastoris can yield large quantity of protein, it is exactly beneficial for us to produce our product in a large scale. Moreover, we let P.pastoris secret the enzyme by adding a secretion peptide so that we do not need to disrupt the organism in order to decrease the consumables of the following production cost.

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.