Team:CCU Taiwan/Description

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Yeast transformation

Enzyme production

Monolignol polymerization

Human practice

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

  In liginin synthesis, monolignols react with two enzymes, peroxidase and laccase. Liggreen, is modelled on Picea abies, which is the most common species used in forestry and possess high content of coniferyl alcohol (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 Pichia pastoris (P. pastoris) (strain: 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

  Literature shows that monolignols forms dimers or oligomers with the assistance of Laccase. After that, through preoxydase, they then start to polymerize, and Liggreen would be produced.
There're three types of bond forms between monolignals: β-β, β-O-4 and β-5.

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.