The role of streptavidin and its variants in catalysis by biotinylated secondary amines

Alexander R. Nödling, Nicolò Santi, Raquel Castillo, Magdalena Lipka-Lloyd, Yi Jin, Louis C. Morrill, Katarzyna Świderek, Vicent Moliner, Louis Y. P. Luk

Research output: Contribution to journalArticlepeer-review


Here, we combine the use of host screening, protein crystallography and QM/MM molecular dynamics simulations to investigate how the protein structure affects iminium catalysis by biotinylated secondary amines in a model 1,4 conjugate addition reaction. Monomeric streptavidin (M-Sav) lacks a quaternary structure and the solvent-exposed reaction site resulted in poor product conversion in the model reaction with low enantio- and regioselectivities. These parameters were much improved when the tetrameric host T-Sav was used; indeed, residues at the symmetrical subunit interface were proven to be critical for catalysis through a mutagenesis study. The use of QM/MM simulations and the asymmetric dimeric variant D-Sav revealed that both Lys121 residues which are located in the hosting and neighboring subunits play a critical role in controlling the stereoselectivity and reactivity. Lastly, the D-Sav template, though providing a lower conversion than that of the symmetric tetrameric counterpart, is likely a better starting point for future protein engineering because each surrounding residue within the asymmetric scaffold can be refined for secondary amine catalysis. This journal is

Original languageEnglish
Pages (from-to)10424-10431
Number of pages8
JournalOrganic & biomolecular chemistry
Issue number47
Publication statusPublished - 15 Nov 2021

Research Beacons, Institutes and Platforms

  • Manchester Institute of Biotechnology


Dive into the research topics of 'The role of streptavidin and its variants in catalysis by biotinylated secondary amines'. Together they form a unique fingerprint.

Cite this