Difference between revisions of "Part:BBa K4881029"

Revision as of 18:58, 8 October 2023

AdhE with AmilCP reporter

The construct starts with a T7 promoter, followed by a ribosome binding site and the adhE gene. adhE encodes for fused acetaldehyde-CoAdehydrogenase and catalyzes the reaction of acetyl-CoA to ethanol in anaerobic fermentation conditions. Lastly, it has AmilCP as a reporter gene that will encode for blue chromoprotein as a visual indicator the bacteria took the plasmid. We made this construct based on the preliminary research as a way to enhance the fermentation process.

Usage and Biology

According to the paper “Metabolic engineering of Escherichia coli for production of mixed-acid fermentation end products”, the main genes involved in the fermentation pathway of E. coli are alcohol dehydrogenase, pyruvate formate lyase, and pyruvate kinase. The image below depicts a diagram of this pathway. The AdhE enzyme sequentially reduces acetyl-CoA to acetaldehyde and then to ethanol under anaerobic conditions. This gene plays a vital role in the fermentation pathway, which is why our team developed a device to express more of this enzyme in E. coli and maximize the amount of ethanol produced in our project.

Sequence and Features

Assembly Compatibility:

10
INCOMPATIBLE WITH RFC[10]
Illegal EcoRI site found at 1
Illegal XbaI site found at 3575
Illegal PstI site found at 203
Illegal PstI site found at 538
Illegal PstI site found at 2104
Illegal PstI site found at 2693
Illegal PstI site found at 3590
12
INCOMPATIBLE WITH RFC[12]
Illegal EcoRI site found at 1
Illegal PstI site found at 203
Illegal PstI site found at 538
Illegal PstI site found at 2104
Illegal PstI site found at 2693
Illegal PstI site found at 3590
Illegal NotI site found at 7
Illegal NotI site found at 3583
21
INCOMPATIBLE WITH RFC[21]
Illegal EcoRI site found at 1
Illegal BglII site found at 570
23
INCOMPATIBLE WITH RFC[23]
Illegal EcoRI site found at 1
Illegal XbaI site found at 3575
Illegal PstI site found at 203
Illegal PstI site found at 538
Illegal PstI site found at 2104
Illegal PstI site found at 2693
Illegal PstI site found at 3590
25
INCOMPATIBLE WITH RFC[25]
Illegal EcoRI site found at 1
Illegal XbaI site found at 3575
Illegal PstI site found at 203
Illegal PstI site found at 538
Illegal PstI site found at 2104
Illegal PstI site found at 2693
Illegal PstI site found at 3590
1000
INCOMPATIBLE WITH RFC[1000]
Illegal SapI.rc site found at 1777

@@ Line 3: / Line 3: @@
 <partinfo>BBa_K4881029 short</partinfo>
-The construct starts with a T7 promoter, followed by a ribosome binding site and the adhE gene. adhE encodes for fused acetaldehyde-CoAdehydrogenase, and catalyzes the reaction of acetyl-CoA to ethanol in anaerobic fermentation conditions. Lastly, it has AmilCP as a reporter gene that will encode for blue chromoprotein as a visual indicator the bacteria took the plasmid. We made this construct based on the preliminary research for our project, as a way to enhance the fermentation process.
+The construct starts with a T7 promoter, followed by a ribosome binding site and the adhE gene. adhE encodes for fused acetaldehyde-CoAdehydrogenase and catalyzes the reaction of acetyl-CoA to ethanol in anaerobic fermentation conditions. Lastly, it has AmilCP as a reporter gene that will encode for blue chromoprotein as a visual indicator the bacteria took the plasmid. We made this construct based on the preliminary research as a way to enhance the fermentation process.
-<!-- Add more about the biology of this part here
 ===Usage and Biology===
+According to the paper “Metabolic engineering of Escherichia coli for production of mixed-acid fermentation end products”, the main genes involved in the fermentation pathway of E. coli are alcohol dehydrogenase, pyruvate formate lyase, and pyruvate kinase. The image below depicts a diagram of this pathway. The AdhE enzyme sequentially reduces acetyl-CoA to acetaldehyde and then to ethanol under anaerobic conditions. This gene plays a vital role in the fermentation pathway, which is why our team developed a device to express more of this enzyme in E. coli and maximize the amount of ethanol produced in our project.
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