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The role of streptavidin and its variants in catalysis by biotinylated secondary amines
Authors:
Alexander R.
Nödling
(Cardiff University)
,
Nicolò
Santi
(Cardiff University)
,
Raquel
Castillo
(Universitat Jaume I)
,
Magdalena
Lipka-Lloyd
(Cardiff University)
,
Yi
Jin
(Cardiff University)
,
Louis C.
Morrill
(Cardiff University)
,
Katarzyna
Świderek
(Universitat Jaume I)
,
Vicent
Moliner
(Universitat Jaume I)
,
Louis Y. P.
Luk
(Cardiff University)
Co-authored by industrial partner:
No
Type:
Journal Paper
Journal:
Organic & Biomolecular Chemistry
, VOL 19
, PAGES 10424 - 10431
State:
Published (Approved)
Published:
November 2021
Diamond Proposal Number(s):
14843
,
18812

Abstract: 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.
Diamond Keywords: Enzymes
Subject Areas:
Biology and Bio-materials,
Chemistry
Instruments:
I03-Macromolecular Crystallography
,
I04-Macromolecular Crystallography
Added On:
14/03/2022 08:42
Documents:
d1ob01947c.pdf
Discipline Tags:
Biochemistry
Catalysis
Chemistry
Structural biology
Life Sciences & Biotech
Technical Tags:
Diffraction
Macromolecular Crystallography (MX)