Part:BBa_K1991009

Pcons-RBS-LO-AOX2-His

DISPLAYING AOX ENZYME ON THE CELL SURFACE OF E. COLI

A protein can be displayed on the cell surface of E. coli by fusing to Lpp-OmpA. Lipoprotein ([http://www.uniprot.org/uniprot/P69776 Lpp]) is a major outer membrane of E. coli which interacts with the peptidoglycan to maintain the structure and function of cell membrane. Another transmembrane protein called outer membrane protein A ([http://www.uniprot.org/uniprot/P0A910 OmpA]) is involved in bacterial conjugation and phage infection. A study showed that Lpp-OmpA hybrid can direct the heterologous protein GFP to the external surface of E. coli (Enzyme Microb Technol. 2001). Bacillus lipase (J Microbiol. 2014) and Fungi xylanase (Curr Microbiol. 2015) were demonstrated to be displayed on the cell surface of E. coli and maintained the functional enzyme activities. In iGEM 2016 MINGDAO's project, AOX (alcohol oxidase) was displayed on bacterial surface by fusing with Lpp-OmpA, which was proved by protein analysis, enzyme activity assay and TMB assay

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NheI site found at 7
  Illegal NheI site found at 30
  Illegal NotI site found at 2498
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BamHI site found at 497
  Illegal XhoI site found at 2507
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 452
  
• 1000
  COMPATIBLE WITH RFC[1000]