Part:BBa_K4583053

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.

The pathway of PHB synthetsis is conflict with TCA cycle.

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.

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.

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