Composite

Part:BBa_K4146106

Designed by: Yazhao Liu   Group: iGEM22_BUCT   (2022-09-16)
Revision as of 15:51, 11 October 2022 by Ishi (Talk | contribs)


The complete expression box used for expressing caffeine.

The complete expression box used for expressing caffeine. 


  Synthesis.jpeg



 Pathway111.png

Our project is designed to help people improve their scalp microecology. After considering many comprehensive factors, we believe that E. coli Nissle 1917 is a safe, easy to operate and easy to absorb non-esterified fatty acids chassis. So all of these genes are in E. coli Nissle 1917.

Standard curve of thopromine.jpeg

Standard curve of caffeine.jpeg

Production of theobromine.jpeg

Production of caffeine.jpeg

The gene of TcAncCS1 is derived from Cocobroma.Xanthine can be converted to 3-methylxanthine (3-mX) by TcAncCS1 from Theobroma. TcCS2 from C. sinensis has a higher propensity to recognize 3-mX and can convert 3-mX to theobromine. We found that there are many ways to synthesis caffeine. The main caffeine biosynthetic pathway is a sequence consisting of xanthosine→7-methylxanthosine→7-methylxanthine→theobromine→caffeine. Genes encoding N-methyltransferases involved in three of these four reactions have been isolated and the molecular structure of N-methyltransferases investigated and a sequence consisting of xanthine →3-methylxanthine → theobromine → caffeine. In three of the four reactions, the gene encoding N-methyltransferase was involved.We chose the second way. CS stands for caffeine synthase, while the TcAncCS1 ancestral gene is derived from Cocobroma, the duplication of TcAncCS1 at node O leads to TcAncCS2, but the kinetic parameters of TcAncCS2 and its progeny enzymes predict that experts will be favored because the 3 methyl Xanthine does not compete for active site binding. We introduced this plasmid gene to facilitate the synthesis of caffeine in engineered strains, thereby regulating the balance of scalp oil.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 2479
    Illegal PstI site found at 2602
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 2479
    Illegal PstI site found at 2602
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 2479
    Illegal PstI site found at 2602
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 2479
    Illegal PstI site found at 2602
    Illegal NgoMIV site found at 643
    Illegal NgoMIV site found at 1741
  • 1000
    COMPATIBLE WITH RFC[1000]


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