Part:BBa_K5089011:Design

ABO(H243R)-CsgA-LCI

Assembly Compatibility:

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

  Illegal BamHI site found at 812
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 831
  Illegal NgoMIV site found at 977
  Illegal AgeI site found at 335
  
• 1000
  COMPATIBLE WITH RFC[1000]

Design Notes

This expression system is codon optimized for expression in E.coli

Source

ABO(H243R) is an α/β hydrolase mutant with a H243R mutation to improve enzyme stability at acidic pH. Wildtype enzyme is native to Alcanivorax borkumensis, gene number 2449 [1]. LCI (Liquid Chromatography peak I) is a binding peptide for polystyrene, polypropylene and polylactic acid [2]. CsgA is a protein component of E.coil biofilms, which can be used as an anchor to surface display proteins [3].

References

1. Chen, A., Xu, T., Ge, Y., Wang, L., Tang, W., & Li, S. 2019. Hydrogen-bond-based protein engineering for the acidic adaptation of Bacillus acidopullulyticus pullulanase. Enzyme and Microbial Technology, 124, 79–83. https://doi.org/10.1016/j.enzmictec.2019.01.010

2. Lu, Y., Hintzen, K.-W., Kurkina, T., Ji, Y., and Schwaneberg, U. 2023. Directed Evolution of Material Binding Peptide for Polylactic Acid-specific Degradation in Mixed Plastic Wastes. ACS Catalysis, 13(19), 12746–12754. https://doi.org/10.1021/acscatal.3c02142

3. Fei Li, Luona Ye, Longyu Zhang, Xiaoyan Li, Xiaoxiao Liu, Jiarui Zhu, Huanhuan Li, Huimin Pang, Yunjun Yan, Li Xu, Min Yang, Jinyong Yan, Design of a genetically programmed barnacle-curli inspired living-cell bioadhesive, Materials Today Bio, Volume 14, 2022.