Team:Utah State/Results/MedalRequirements

From 2014.igem.org

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<li> Produce stain-fighting enzymes in E. coli </li>
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<li> Produce stain-fighting enzymes in <i>E. coli</i> </li>
<li> Show that enzymes can help remove stains </li>
<li> Show that enzymes can help remove stains </li>
<li> Immobilize enzymes to bioplastic</li>
<li> Immobilize enzymes to bioplastic</li>
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1. To produce stain-fighting enzymes in E. coli, we need to genetically engineer our bacteria using genes that code for the synthesis of our three enzymes: cellulase, amylase, and chlorophyllase. Each construct should include a histidine tag for protein purification. After enzymes are purified, assays with each enzyme’s substrate should be conducted in order to determine if the enzyme is active or not.     
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1. To produce stain-fighting enzymes in <i>E. coli</i>, we need to genetically engineer our bacteria using genes that code for the synthesis of our three enzymes: cellulase, amylase, and chlorophyllase. Each construct should include a histidine tag for protein purification. After enzymes are purified, assays with each enzyme’s substrate should be conducted in order to determine if the enzyme is active or not.     
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2. In order to show that enzymes can remove stains, multiple assays will be conducted to demonstrate each enzyme’s ability to degrade its specific substrate i.e. cellulase degrading cellulose in CMC plates, amylase degrading the starch in starch plates, and chlorophyllase cleaving the phytol tail of chlorophyll to make it water soluble. These assays will provide a proof of concept; pairing this proof of concept with background literature reviews will help demonstrate that enzymes can in fact remove stains.  
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2. In order to show that enzymes can remove stains, multiple assays will be conducted to demonstrate each enzyme’s ability to degrade its specific substrate (i.e. cellulase degrading cellulose in CMC agar plates, amylase degrading starch in starch agar plates, and chlorophyllase cleaving the phytol tail of chlorophyll to make it water soluble. These assays will provide a proof of concept; pairing this proof of concept with background literature reviews will help demonstrate that enzymes can in fact remove stains.  
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3. To immobilize enzymes to bioplastic, we can use previous research done at our university regarding the production of bioplastic in E. coli. Instead of attaching GFP to the bioplastic producing genes, we can replace GFP’s spot with each of our enzymes. Can also use chemical immobilization as a proof of concept.  
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3. To immobilize enzymes to bioplastic, we can build upon previous iGEM team's work regarding the production of bioplastic in <i>E. coli</i>. After using green fluorescent protein (GFP) as a reporter to illustrate attachment to the bioplastic granules inside the cell, we can go forward with the same strategy for each of our enzymes. To further solidify our end goal, we can chemically immobilize our BioBrick-produced enzymes to PVC plastic and show that the enzymes retained their efficacy after attachment (chemical immobilization would result in a covalent bond between enzyme and activated PVC plastic, the same type of bond that would occur in our biological immobilization strategy).
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4. The objective of creating a reusable bioplastic laundry treatment is to make clothes last longer as well as reduce waste. To create this treatment, we will have to prove that we can immobilize our laundry detergent enzymes to bioplastic. Second, we will have to be able to produce a large amount of immbolized enzyme bioplastic. Last, we will take that bioplastic and either make a spherical structure out of it or apply it to a scrubbing brush.
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4. The objective of creating a reusable bioplastic laundry treatment is to reduce the quantity of harsh detergents, minimize wastes of non-reusable "free" enzymes, and improve upon the removal of tough clothing stains. To create this treatment, we first must prove that we can immobilize our stain-fighting enzymes to bioplastic while maintaining their efficacy. Second, we must be able to produce a sufficient amount of enzyme-immbolized bioplastic to allow for rigorous testing. We can then take this enzyme-immobilized bioplastic and construct an additive device to traditional laundry cycles that can be reused in multiple washes. Furthermore, we can use the stain-fighting bioplastic to surface coat scrubbing brushes and buckets for use in under-developed countries and for those who cannot afford traditional laundry machines.
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5. To win our 6th straight gold medal, we will need to make sure to follow the gold medal requirements as well as making sure our project is either novel or improves upon existing iGEM or research projects.  
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5. To win our 6th straight Gold Medal, the Utah State iGEM team will need to continue to go above and beyond the requirements necessary for a Gold Medal. We will need to ensure that our project is both novel,  and helps build upon previous iGEM team projects.  
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6. In order to win best manufacturing project, we have to make sure that our project exceeds expectations and provides a novel approach to an ongoing problem.   
6. In order to win best manufacturing project, we have to make sure that our project exceeds expectations and provides a novel approach to an ongoing problem.   

Revision as of 04:19, 17 October 2014

Medal Requirements

USU 2014iGem2014;

Goals

Goals

  1. Produce stain-fighting enzymes in E. coli
  2. Show that enzymes can help remove stains
  3. Immobilize enzymes to bioplastic
  4. Create a reusable bioplastic laundry treatment
  5. Win 6th straight gold medal
  6. Win best manufacturing project

1. To produce stain-fighting enzymes in E. coli, we need to genetically engineer our bacteria using genes that code for the synthesis of our three enzymes: cellulase, amylase, and chlorophyllase. Each construct should include a histidine tag for protein purification. After enzymes are purified, assays with each enzyme’s substrate should be conducted in order to determine if the enzyme is active or not.

2. In order to show that enzymes can remove stains, multiple assays will be conducted to demonstrate each enzyme’s ability to degrade its specific substrate (i.e. cellulase degrading cellulose in CMC agar plates, amylase degrading starch in starch agar plates, and chlorophyllase cleaving the phytol tail of chlorophyll to make it water soluble. These assays will provide a proof of concept; pairing this proof of concept with background literature reviews will help demonstrate that enzymes can in fact remove stains.

3. To immobilize enzymes to bioplastic, we can build upon previous iGEM team's work regarding the production of bioplastic in E. coli. After using green fluorescent protein (GFP) as a reporter to illustrate attachment to the bioplastic granules inside the cell, we can go forward with the same strategy for each of our enzymes. To further solidify our end goal, we can chemically immobilize our BioBrick-produced enzymes to PVC plastic and show that the enzymes retained their efficacy after attachment (chemical immobilization would result in a covalent bond between enzyme and activated PVC plastic, the same type of bond that would occur in our biological immobilization strategy).

4. The objective of creating a reusable bioplastic laundry treatment is to reduce the quantity of harsh detergents, minimize wastes of non-reusable "free" enzymes, and improve upon the removal of tough clothing stains. To create this treatment, we first must prove that we can immobilize our stain-fighting enzymes to bioplastic while maintaining their efficacy. Second, we must be able to produce a sufficient amount of enzyme-immbolized bioplastic to allow for rigorous testing. We can then take this enzyme-immobilized bioplastic and construct an additive device to traditional laundry cycles that can be reused in multiple washes. Furthermore, we can use the stain-fighting bioplastic to surface coat scrubbing brushes and buckets for use in under-developed countries and for those who cannot afford traditional laundry machines.

5. To win our 6th straight Gold Medal, the Utah State iGEM team will need to continue to go above and beyond the requirements necessary for a Gold Medal. We will need to ensure that our project is both novel, and helps build upon previous iGEM team projects.

6. In order to win best manufacturing project, we have to make sure that our project exceeds expectations and provides a novel approach to an ongoing problem.