2 publications

2 publications

Construction of Robust Bio-Nanotubes using the Controlled Self-Assembly of Component Proteins of Bacteriophage T4

Ueno, T.

Small 2010, 6, 1873-1879, 10.1002/smll.201000772

The synthesis of a robust bio‐nanotube consisting of the β‐helical tubular component proteins of bacteriophage T4 is described. The crystal structure indicates that it has a well‐defined nanoscale length of 10 nm as a result of the head‐to‐head dimerization of β‐helices. Surprisingly, the tube assembly has high thermal stability, high tolerance to organic solvents, and a wide pH‐stability range.


Metal: Cu
Ligand type: Flavin
Host protein: [(gp5βf)3]2
Anchoring strategy: Lysine-succinimide
Optimization: ---
Reaction: Cycloaddition
Max TON: ---
ee: ---
PDB: ---
Notes: ---

Directed Evolution of a Cp*RhIII‐Linked Biohybrid Catalyst Based on a Screening Platform with Affinity Purification

Hayashi, T; Onoda, A.

ChemBioChem 2021, 22, 679-685, 10.1002/cbic.202000681

Directed evolution of Cp*RhIII-linked nitrobindin (NB), a biohybrid catalyst, was performed based on an in vitro screening approach. A key aspect of this effort was the establishment of a high-throughput screening (HTS) platform that involves an affinity purification step employing a starch-agarose resin for a maltose binding protein (MBP) tag. The HTS platform enables efficient preparation of the purified MBP-tagged biohybrid catalysts in a 96-well format and eliminates background influence of the host E. coli cells. Three rounds of directed evolution and screening of more than 4000 clones yielded a Cp*RhIII-linked NB(T98H/L100K/K127E) variant with a 4.9-fold enhanced activity for the cycloaddition of acetophenone oximes with alkynes. It is confirmed that this HTS platform for directed evolution provides an efficient strategy for generating highly active biohybrid catalysts incorporating a synthetic metal cofactor.


Metal: Rh
Ligand type: Cp
Host protein: Nitrobindin (Nb)
Anchoring strategy: Covalent
Optimization: Genetic
Reaction: Cycloaddition
Max TON: ---
ee: ---
PDB: ---
Notes: ---