TTE REU Final presentation
-
Upload
kamaria-kermah -
Category
Documents
-
view
202 -
download
0
Transcript of TTE REU Final presentation
![Page 1: TTE REU Final presentation](https://reader035.fdocuments.in/reader035/viewer/2022062900/58e783ae1a28abe7528b591b/html5/thumbnails/1.jpg)
ENGINEERING SIGNAL SEQUENCES ON THE 1,2-PROPANEDIOL UTILIZATION
MICROCOMPARTMENT
Kamaria Ashley KermahMentor: Chris JakobsonDanielle Tullman-Ercek
Department of Chemical and Biomolecular Engineering
July 28, 2015
Purified Microcompartment
![Page 2: TTE REU Final presentation](https://reader035.fdocuments.in/reader035/viewer/2022062900/58e783ae1a28abe7528b591b/html5/thumbnails/2.jpg)
ENGINEERING BACTERIAL CELLS
• Why?
o Ubiquitous
o Easily Replicated
o DNA Easily Manipulated
•How?
o Manipulating the genetic code
![Page 3: TTE REU Final presentation](https://reader035.fdocuments.in/reader035/viewer/2022062900/58e783ae1a28abe7528b591b/html5/thumbnails/3.jpg)
1,2-PROPANEDIOL UTILIZATION MICROCOMPARTMENT (PDU MCP)
• Proteinacious Organelle
• Protection and Efficiency
1,2-propanediol
Carbon
![Page 4: TTE REU Final presentation](https://reader035.fdocuments.in/reader035/viewer/2022062900/58e783ae1a28abe7528b591b/html5/thumbnails/4.jpg)
METABOLIC PATHWAY
• Salmonella enterica Pathogenicity in
human gut 1,2-propanediol
propionaldehyde
propionyl-CoA
PduCDE
PduP
PduW
propionateATP
ADP
![Page 5: TTE REU Final presentation](https://reader035.fdocuments.in/reader035/viewer/2022062900/58e783ae1a28abe7528b591b/html5/thumbnails/5.jpg)
PDU CDE COMPLEX
• Enzyme localization
• N-terminus of PduD
• Alpha helix
• Hydrophobic and hydrophilic
faces
![Page 6: TTE REU Final presentation](https://reader035.fdocuments.in/reader035/viewer/2022062900/58e783ae1a28abe7528b591b/html5/thumbnails/6.jpg)
6
HOMOLOGOUS RECOMBINATIONHOW DOES IT WORK?
B.
![Page 7: TTE REU Final presentation](https://reader035.fdocuments.in/reader035/viewer/2022062900/58e783ae1a28abe7528b591b/html5/thumbnails/7.jpg)
7
METHODSGrowth on 1,2-
propanediol
[G. Cannon 2001. Microcompartments in Prokaryotes doI: 10.1128/AEM.67.12.5351-5361.2001]
Homologous Recombination
Sequencing Reaction
TEM Microscopy
![Page 8: TTE REU Final presentation](https://reader035.fdocuments.in/reader035/viewer/2022062900/58e783ae1a28abe7528b591b/html5/thumbnails/8.jpg)
RESULTS AND ANALYSIS
Sucrose Sensitivity Confirmation of gene size
~5kb
Amplification of pduD gene
1kb
0.5kb
Sucrose Resistance
![Page 9: TTE REU Final presentation](https://reader035.fdocuments.in/reader035/viewer/2022062900/58e783ae1a28abe7528b591b/html5/thumbnails/9.jpg)
FUTURE APPLICATIONS
• Engineering microcompartments for industrial use
o Nanobioreactors
o Antibiotics Replace PduD N-terminal with PduP
signal peptide
Increasing size of microcompartments and number of enzymes to increase titer
Localization of co-factors and heterologous enzymes into MCP
MCP formation in yeast