Team:CCU Taiwan/Demonstrate

DEMONSTRATION

Yeasts & Enzymes

  In terms of gene design, our goal is mainly the production of plant enzymes. Therefore, we demonstrated the growth curve of the yeast and ensure the efficiency of the production of enzymes.

Figure1: ZeocinTM selection



Figure2: Growth curve data.



Figure3: Growth curve measurement: Lac1.



Figure4: Growth curve measurement: Px16.



Figure5: Growth curve measurement: Px18.



His4 experiment

  In our experiments, we have successfully constructed pGAPZ A containing His4 vectors. As of yet, we have just successfully transformed pGAPZ A_His4_Px18 vector into P. pastoris. In the future, we will do some further researches on this experiment.

Figure6: Plasmid digest (EcoRI, Agel) after miniprep



Figure7: Plasmid digest(SalI) check

Figure8: pGAPZ A_His4_Px18 selection with SD plate

Properties analysis

  For our production methods, we refer to the current material production and get some engineering conditions from the company. Through these engineering conditions, we have designed some modeling and production line. We hope that LIGGREEN will be more in line with business needs. We hope that LIGGREEN will have a place in the market.
  In terms of engineering properties of LIGGREEN, we also completed two important parts.



PART 1.  Modeling

Spontaneous reaction of monolignols & calculation of Flory-Stockmayer Theory of polymerization

  In our modeling, we constructed a theoretical calculation for our material. Based on published literature, we used Spartan to determine the spontaneous basis for our bond reaction, and then Flory-Stockmayer theory to provide a theoretical estimate of the polymerization. Through the establishment of the model, we can get theoretical data on LIGGREEN. In patent applications and product improvements, we can improve on this estimate through theoretical calculations and material measurements.

Figure9: Three Bonds based on Spartan Analysis



PART 2.  Structure analysis

  We analyzed the structure of our product. By UV-Visible we proved that the reaction has occurred and the linkage in our product is what we have expected. Also form other analysis, we learned more about the characteristics of our products.



Figure10: 1st reaction experiment result(coniferyl alcohol react with laccase for 18hr at 25°C)

Figure11: 2nd reaction experiment result(coniferyl alcohol react with laccase for 18hr at 25°C)




PART 3.  Production line

  We have designed a production line to produce LIGGREEN. Using existing fermentation tank systems and reaction tank assemblies on the market, we can construct our production line.



Figure12: Production line



Safety

  For our production methods, we have designed an effective biological control. In the production line, we have a filter between the fermentation tank and the reaction tank. In addition, our production line will be heat-sterilized after the reaction tank. We have done experiments to prove that our yeast will not cause a biological crisis.


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Market analysis


  We have already done a market survey of LIGGREEN, to match the needs of the commodity market. We have built our own business models and various analyses, such as SWOT analysis, STP. Furthermore, we have visited several companies that produce laminated paper cups, to understand their requirements and adjust our product to be suitable for them.(Read more in Entrepreniumship)



  Base on corporate social responsibility, we commit to produce environmental friendly product. Moreover, to bring more welfare to the society, we proposed two policies about food packaging safety and recycling regulations. We consult county councilor to understand the progress of establishing policies and put ours into practice. (Read more in Intergrate HP)