Part:BBa_K3143341

SpyCatcher-Mercury Binding Protein

Mercury Binding Protein fused to the SpyCatcher domain can be covalently attached onto the biofilm displaying SpyTag for heavy metal adsorption^[1].

Characterization

We first expressed SpyCatcher-mecury binding protein(MBP) in E.coli BL21 Cell, then lysed cells and collected the SpyCatcher-MBP protein in the supernatant. We added a certain amount of SpyCatcher-MBP protein to 1mL over-night bacterial culture medium of CsgA–INP-SpyTag with 10^-5μM of Hg²⁺ concentration, reacting for two hours. Then, we centrifuged the reaction system and the supernatant was test using ICP-MS. The decrease of Hg²⁺ concentration level indicated the amount of Hg²⁺ captured using our adsorber cells.

As shown in the results in Fig. 1b, 1mL NEZHA can absorb more than 50% of 10^-5 μM Hg²⁺ in 120 minutes. That is to say, a NEZHA cell can absorb about 6000-10000 Hg ions. (Assume 1mL of bacteria contains 10⁹ bacteria).

Figure 1: A PAGE Gel showed that SpyCatcher-mercury binding protein was succefully expressed and collected B ICP-MS results of the MEZHA mercury adsorption

Reference

1. Nguyen P Q, Botyanszki Z, Tay P K R, et al. Programmable biofilm-based materials from engineered curli nanofibres[J]. Nature communications, 2014, 5: 4945.

Sequence and Features

Assembly Compatibility:

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

  Illegal BamHI site found at 547
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]