Part:BBa_K4146507

pCS-aroGfbr

We hope to use 2-PE as an example to give E. coli Nissle 1917 the ability to sweeten hair in addition to improving scalp microecology. We use the natural phenylalanine synthesis pathway of E. coli to synthesize 2-PE. aroG was used to enhance the accumulation of a key intermediate, DAHP. aroG, as a metabolic pathway gene that has attracted much attention for many years, has always been a worthy object of study. We believe that its feedback inhibition may affect the synthesis of 2-phenylethyl alcohol. We use this Mutant Feedback-Resistant Version of aroG for to resisting feedback inhibition and improving enzyme activity. This is the composite of our Improvement of Standard Biological Parts.

Fig.1 The standard curve for determination of 2-phenylethanol yield

In the standard curve, as shown in Fig.1,the ordinate is the peak area and the abscissa is the marker concentration.The mobile phase of high performance liquid chromatography is trifluoroacetic acid diluted 1000 times.In our experiment, we took two points, 500 mg/L and 1000 mg/L, as well as zero point. The R square fitted by the standard curve is 1, which shows that the experimental results are fitted very well, and the error can be almost ignored, that is, the measured output is relatively accurate.

Fig.2 The experimental 2-phenylethanol yield

Our team successfully constructed a pathway for the synthesis of 2-phenylethanol in Escherichia coli. When cultured for 60 hours, the maximum concentration of 2-phenylethanol was 67.16mg/L.After 60 hours,the concentration of 2-phenylethanol decreases because of toxicity of 2-PE accumulated in E. coli and metabolism.

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal EcoRI site found at 450
  Illegal EcoRI site found at 665
  Illegal XbaI site found at 87
  Illegal XbaI site found at 941
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal EcoRI site found at 450
  Illegal EcoRI site found at 665
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal EcoRI site found at 450
  Illegal EcoRI site found at 665
  Illegal BamHI site found at 738
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal EcoRI site found at 450
  Illegal EcoRI site found at 665
  Illegal XbaI site found at 87
  Illegal XbaI site found at 941
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal EcoRI site found at 450
  Illegal EcoRI site found at 665
  Illegal XbaI site found at 87
  Illegal XbaI site found at 941
  
• 1000
  COMPATIBLE WITH RFC[1000]