Difference between revisions of "Team:Edinburgh OG/Demonstrate"

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<p style="text-align:center;"><h1><b>Demonstrate</b></h1>
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Our project's final aim was to create PHBV efficiently from waste or industrial by-products. Additionally, We wanted to reduce the need for downstream processing which is costly environmentally (see our LCA). To demonstrate this we used our bktb/phaCB operon grown on whisky pot ale to create plastic.</p>
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<h1 style="text-align: center;"><strong>Integrated Human Practices</strong></h1>
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<p style="text-align: justify;"> Our project's final aim was to create PHBV efficiently from waste or industrial by-products. Additionally, We wanted to reduce the need for downstream processing which is costly environmentally (see our LCA). To demonstrate this we used our bktb/phaCB operon grown on whisky pot ale to create plastic.</p>
 
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Revision as of 00:37, 18 October 2018

PhagED: a molecular toolkit to re-sensitise ESKAPE pathogens

 

 

 

 

 

Integrated Human Practices

Our project's final aim was to create PHBV efficiently from waste or industrial by-products. Additionally, We wanted to reduce the need for downstream processing which is costly environmentally (see our LCA). To demonstrate this we used our bktb/phaCB operon grown on whisky pot ale to create plastic.

        

Secreation

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Growing on Whisky Pot Ale

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Figure 1. Six 50 mL cultures of phaCB/Bktb in M9 media and 1% glucose were set up with three containing Pot ale and the other three containing water. Colonies were set up to shake at 37 oC overnight for 64 hours.



We were able to grow our E. coli on media containing M9 salts, 1% glucose, and pot ale. Our negative control contained water instead of pot ale. Our cells grew similar in both types of media but grew lower than our model predicted. However, the pot ale did not negatively affect growth.

Figure 2. OD600 of cells grown in M9 media and 1% glucose with and without pot ale. Cells grown in pot ale and without pot ale similar growth curves.



Next we wanted to show that our cells could produce plastic growing in pot ale. Cells were grown for 64 hours and then had their dry cell weight measured before extraction (Figure 3).

Figure 3. Dry cell weight vs mass of plastic extracted of phaCB-Bktb cells grown in M9 media and 1% glucose with and without pot ale.



Figure 4. Melting temperature ranges of extracted plastic. Cultures grown on pot ale have consistant melting temperature ranges.

We were able to identify by GC-MS two of the main products of PHBV depolymerisation and dehydration by sulphuric acid: crotonic acid and 2-pentenoic acid (Xiang et al., 2016; Braunegg et al., 1978). Demonstrating the presence of PHBV in the processed samples Figure X and Figure X.