Part:BBa_K4891014

glk-glf

It is the key part that is responsible for strengthening glucose utilization rate. The phosphotransferase system was replaced by heterogeneously introducing glucose facilitator in strain E. coli ptsG:glk-glf.

Usage and Biology

We engineered the shikimate pathway of E. coli MG1655 to efficiently accumulate SA. We knocked out ptsG, ldhA, adhE, poxB, and pta genes to achieve the production of precursor substance PEP. To increase intracellular E4P content, we overexpressed tktA and talB genes. To enhance product accumulation, we overexpressed aroG, aroB, aroD, and aroE genes, while knocking down aroK and aroL genes to cut off the metabolic flux, thus accomplishing the accumulation of shikimic acid. Besides, a non-phosphorylated pathway, that is, glk and glf genes (mainly by glk-glf integration into the ptsG locus) is introduced to enhance glucose utilization. To achieve the goal above-mentioned, we totally constructed 26 parts this year. See table below:

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal PstI site found at 124
  Illegal PstI site found at 1136
  Illegal PstI site found at 1327
  Illegal PstI site found at 1900
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal PstI site found at 124
  Illegal PstI site found at 1136
  Illegal PstI site found at 1327
  Illegal PstI site found at 1900
  
• 21
  COMPATIBLE WITH RFC[21]
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal PstI site found at 124
  Illegal PstI site found at 1136
  Illegal PstI site found at 1327
  Illegal PstI site found at 1900
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal PstI site found at 124
  Illegal PstI site found at 1136
  Illegal PstI site found at 1327
  Illegal PstI site found at 1900
  Illegal AgeI site found at 421
  Illegal AgeI site found at 1312
  Illegal AgeI site found at 1468
  Illegal AgeI site found at 1480
  Illegal AgeI site found at 2191
  
• 1000
  COMPATIBLE WITH RFC[1000]