Difference between revisions of "Part:BBa K4583053"

Revision as of 14:26, 11 October 2023

PHB production Regulation: PesaS-B0034-gltA-PesaRwt-B0034-PHBcab-PYU16-B0034-SRRz

PesaS-B0034-gltA-PesaRwt-B0034-PHBcab-PYU16-B0034-SRRz

Usage and Biology

Many production processes using microorganisms face the dilemma of conflicting production products and key metabolic pathways. This means that simply introducing product-synthesising genes into engineered bacteria can greatly affect the growth of the microorganism, leading to a situation where production is too low. There are a number of current solutions to this problem. For example, metabolic engineering can regulate metabolic flow using methods such as gene knockdown, promoter replacement, etc. These static strategies are effective for productivity improvement, but are not responsive to changes in the cell or environment. Dynamic control is a favourable solution for the conditional knockdown of essential genes and balances the flow in the metabolic pathway.

This part is about PHB Production Regulation based on an three-layer dynamic regulation model. It dynamically regulate the PHB production by dividing the process into 3 phases: Growth Phase, Production Phase and Product-release Phase. In this way, the cell can firstly grow up and then put their all effort into PHB production. Finally, in the late stationary phase of cell growth, the enigneered bacteria can express lysis gene.

1. Three-layer Dynamic Regulation Model

2. The first and second layer--Growth and production control

Quorum Sensing is a way for cells to regulate downstream gene expression based on their own density. The concentration of the signaling molecule - AHL - secreted by the cell increases as the cell density increases. When the concentration of AHL reaches a certain level, it can bind to the corresponding binding protein and alter the expression of downstream genes.

4. The Third layer--Product-release control

Characterization

1. Protocols

2. The Characterization of PesaS and PesaRwt/PesaRc/PesaRp

3. The Characterization of PesaRwt/PesaRc/PesaRp and PYU3/PYU7/PYU16/PYU92

Summary of Results

Our part provides an example for future iGEM team

Limitations

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
INCOMPATIBLE WITH RFC[21]
Illegal BglII site found at 2740
Illegal BglII site found at 3565
Illegal BamHI site found at 1791
Illegal XhoI site found at 1511
23
COMPATIBLE WITH RFC[23]
25
INCOMPATIBLE WITH RFC[25]
Illegal NgoMIV site found at 2046
Illegal NgoMIV site found at 2117
Illegal NgoMIV site found at 2717
Illegal NgoMIV site found at 3029
Illegal NgoMIV site found at 3308
Illegal NgoMIV site found at 3960
Illegal NgoMIV site found at 3982
1000
INCOMPATIBLE WITH RFC[1000]
Illegal BsaI site found at 5826
Illegal SapI.rc site found at 769

@@ Line 5: / Line 5: @@
 PesaS-B0034-gltA-PesaRwt-B0034-PHBcab-PYU16-B0034-SRRz
 ==Usage and Biology==
-Many production processes using microorganisms face the dilemma of conflicting production products and key metabolic pathways. This means that simply introducing product-synthesising genes into engineered bacteria can greatly affect the growth of the microorganism, leading to a situation where production is too low. There are a number of current solutions to this problem. For example, metabolic engineering can regulate metabolic flow using methods such as gene knockdown, promoter replacement, etc. These static strategies are effective for productivity improvement, but are not responsive to changes in the cell or environment.
+Many production processes using microorganisms face the dilemma of conflicting production products and key metabolic pathways. This means that simply introducing product-synthesising genes into engineered bacteria can greatly affect the growth of the microorganism, leading to a situation where production is too low. There are a number of current solutions to this problem. For example, metabolic engineering can regulate metabolic flow using methods such as gene knockdown, promoter replacement, etc. These static strategies are effective for productivity improvement, but are not responsive to changes in the cell or environment. Dynamic control is a favourable solution for the conditional knockdown of essential genes and balances the flow in the metabolic pathway.
-This part is about PHB Production Regulation based on an three-layer dynamic regulation model. It successfully regulate the PHB production by dividing the process into 3 phases: Growth Phase, Production Phase and Product-release Phase. In this way, the cell factory can firstly grow
+This part is about PHB Production Regulation based on an three-layer dynamic regulation model. It dynamically regulate the PHB production by dividing the process into 3 phases: Growth Phase, Production Phase and Product-release Phase. In this way, the cell can firstly grow up and then put their all effort into PHB production. Finally, in the late stationary phase of cell growth, the enigneered bacteria can express lysis gene.
 ===1. Three-layer Dynamic Regulation Model===
 ===2. The first and second layer--Growth and production control===