Bacterial Cell Display as a Robust and Versatile Platform for Engineering Low-Affinity Ligands and Enzymes

Autor: Eszter Csibra, Vitor B. Pinheiro, Marleen Renders
Rok vydání: 2020
Předmět:
SELECTION
Biochemistry & Molecular Biology
bacterial cell display
DIRECTED EVOLUTION
Chemistry
Medicinal
Bioengineering
Computational biology
010402 general chemistry
Ligands
01 natural sciences
Biochemistry
DNA-binding protein
Bacterial cell structure
BETA-LACTAMASE
Xenobiotics
Low affinity
TERMINAL PROTEIN
Nucleic Acids
Humans
Pharmacology & Pharmacy
XNA molecular biology
directed evolution
Molecular Biology
ANTIBODY LIBRARIES
chemistry.chemical_classification
Bacterial display
Science & Technology
Alkyl and Aryl Transferases
Bacteria
Full Paper
010405 organic chemistry
Organic Chemistry
PHI-29 DNA-REPLICATION
AUTODISPLAY
Robustness (evolution)
Full Papers
Directed evolution
0104 chemical sciences
Enzyme
chemistry
SURFACE DISPLAY
Nucleic acid
Molecular Medicine
P16.7
Life Sciences & Biomedicine
SYSTEM
Zdroj: Chembiochem
ISSN: 1439-7633
Popis: Directed evolution has been remarkably successful at expanding the chemical and functional boundaries of biology. That progress is heavily dependent on the robustness and flexibility of the available selection platforms, given the significant cost to (re)develop a given platform to target a new desired function. Bacterial cell display has a significant track record as a viable strategy for the engineering of mesophilic enzymes, as enzyme activity can be probed directly and free from interference from the cellular milieu, but its adoption has lagged behind other display‐based methods. Herein, we report the development of SNAP as a quantitative reporter for bacterial cell display, which enables fast troubleshooting and the systematic development of the display‐based selection platform, thus improving its robustness. In addition, we demonstrate that even weak interactions between displayed proteins and nucleic acids can be harnessed for the specific labelling of bacterial cells, allowing functional characterisation of DNA binding proteins and enzymes, thus making it a highly flexible platform for these biochemical functions. Together, this establishes bacterial display as a robust and flexible platform, ideally suited for the systematic engineering of ligands and enzymes needed for XNA molecular biology.
Displaying enzymes and low‐affinity binding proteins on the surface of the bacterial cell: A SNAP reporter together with the function‐dependent labelling of the cell with cholesterol‐linked DNA, allows systematic optimisation of cell display, making it a robust and flexible platform for the systematic evolution of XNA molecular biology.
Databáze: OpenAIRE
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