Part:BBa_K2684005

SpyCatcher-CotA

SpyCatcher fused with CotA by 2xGGGGS linker

Sequence and Features

Assembly Compatibility:

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

  Illegal XhoI site found at 514
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 1903
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal SapI.rc site found at 841
  

This part could be used to produce CotA laccase fused with SpyCatcher (BBa_K2684005). Team SHSBNU_China designed this part to improve the the enzyme activity of traditional CotA. When applying the gene CotA without SpyCatcher, the laccase cannot be suspended onto the biofilm created by CsgA-Spytag. Therefore, there will not be significant improvement in enzyme activity. However, while applying with CsgA-Spytag, the enzyme activity dramatically increased, as shown in the graph below.

The more *, the larger difference between each group

Some experiments showed that, while using singly, the existence of SpyCatcher actually result in a decrease in enzyme activity compared to CotA wild type, as shown in the graph below.

The problem might be:
1. The inserted gene Spycatcher affected expression of laccase.
2. The extra structure of CotA laccase result in a decrease in enzyme activity

For more experimental details, please visit http://2018.igem.org/Team:SHSBNU_China/Experiments