Part:BBa_K1921012

PETase+BrkA

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 1297
  Illegal NgoMIV site found at 1726
  Illegal NgoMIV site found at 2362
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 2330
  

Usage

A new autotransportermediated bacterial surface display system BrkAu to Display based on the structure of BrkA. The unique characteristic of BrkA is that the cleaved passenger domain remains adhered on the bacterial surface, which is conducive to keep the displayed exogenous proteins associated onto the cell, thus, the functionalized bacteria can be recycled/re-used conveniently.

Biology

N-terminal. Autotransporter BrkA (Bordetella serum-resistance killing protein A )is from Bordetella pertussis,PETase is the passenger between the signal peptide and the anchor.

Reference

[1] Fang S, Pang X, Tian X, et al. BrkAutoDisplay: functional display of multiple exogenous proteins on the surface of Escherichia coli, by using BrkA autotransporter[J]. Microbial Cell Factories, 2014, 14(1):1-12.

Pre-expression

Figure 1.This is the pre-expression using E.coli BL21 in different inducing condition.

Surface display HPLC results

Figure 2.Relative enzyme activity of engineering bacteria E.coli(BL21)/pET22b(+)Brk when induced at 16℃ and 25 ℃ with 0.02mM IPTG.And the last two were induced with 0.09mM IPTG.