|
Part number |
Part name |
Contribution type |
|
BBa_K4929003 |
PAO-815-AocⅠ |
New composite part |
|
BBa_K4929004 |
PAO-815-FAD-AocⅠ |
New composite part |
|
FAD |
New basic part |
|
|
pAO-815 |
New basic part |
|
|
AocI |
New basic part |
|
Name |
Description |
Contribution type |
|
Successfully reduce biogenic amines. |
The two plasmids successfully reduce the biogenic amines. |
Reduce biogenic amines. |
1. Part Contributions:
In fermentation processes, yeast and other microorganisms will produce what is known as biogenic amines. It has the potential health risks to human bodies. Our goal is to reduce this amine inside alcohol. To adequately address this issue, we aimed to enhance the safety of alcoholic beverages by genetically modifying yeast to express amine oxidase, an enzyme capable of breaking down biogenic amines.
To achieve this, we designed two plasmids: one carrying the AocI gene, which encodes the amine oxidase enzyme, and the other carrying the FAD gene, facilitates the expression of AocI.
l pAO815-AocI
As the figure shown below, we have pAO815-AocI, where pAO815 is a frequently used isotopic carrier in Pichia pastoris. It serves as a vehicle for expressing the target protein within the yeast.
To ensure proper expression of foreign genes within the yeast, a promoter is required. The AOX1 promoter fulfills this crucial role.
It starts the plasmid and ends with the AOX1 terminator to the cycle of the plasmid.
AocI (BBa_K4929002) is the main part of our design which will express amine oxidase so as to reduce the amine level during yeast fermentation.
For part pAO815-AocI, we have obtained the protein with SDS-PAGE result. In addition, we studied the expression curve of the protein with various concentrations of methanol.
SDS PAGE (left) and the expression level against different methanol concentrations (right) of AocI protein in pAO815-AocI plasmid
l pAO815-AocI-FAD
Compared with pAO815-AocI, pAO815-AocI-FAD is Constructed with FAD gene, which will code to express histamine dehydrogenase so as to facilitate the expression of the AocI.
In order to activate the FAD particle, we bring in another promoter, ADH1 promoter.
For part pAO815-AocI-FAD, we have also obtained the protein with SDS-PAGE result and studied the expression curve of the protein with various concentrations of methanol.
SDS PAGE (left) and the expression level against different methanol concentrations (right) of AocI protein in pAO815-AocI-FAD plasmid
2. Other Contribution:
Both of these plasmids aim to reduce the biogenetic anime inside the alcohol. The difference between them is that one has an extra gene that will make the AocI perform better. In the future, our team plans to put these into the process of fermentation, making alcohol much safer to ingest into our system and not losing its original structure of taste and texture. We think that this innovation in the market of alcohol will make a significant rise in the sale of any products.