Self-Assembly of Amyloid Fibrils That Display Active Enzymes.
Enzyme immobilization is an important strategy to enhance the stability and recoverability of enzymes and to facilitate the separation of enzymes from reaction products. However, enzyme purification followed by separate chemical steps to allow immobilization on a solid support reduces the efficiency and yield of the active enzyme. Here we describe polypeptide constructs that self-assemble spontaneously into nanofibrils with fused active enzyme subunits displayed on the amyloid fibril surface. We measured the steady-state kinetic parameters for the appended enzymes in situ within fibrils and compare these with the identical protein constructs in solution. Finally, we demonstrated that the fibrils can be recycled and reused in functional assays both in conventional batch processes and in a continuous-flow microreactor.
National Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences 15 Datun Road, Chaoyang District, Beijing 100101 (China) ; Department of Chemistry, University of Cambridge Lensfield Road, Cambridge CB2 1EW (UK) ; University of the Chinese Academy of Sciences 19 A Yuquanlu, Shijingshan District, Beijing 100049 (China).
Xiao-Ming
XM
Zhou
National Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences 15 Datun Road, Chaoyang District, Beijing 100101 (China).
Aiman
A
Entwistle
National Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences 15 Datun Road, Chaoyang District, Beijing 100101 (China).
Hong
H
Zhang
National Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences 15 Datun Road, Chaoyang District, Beijing 100101 (China) ; Department of Biochemistry, University of Cambridge Tennis Court Road, Cambridge CB2 1QW (UK).
Antony P
AP
Jackson
Department of Chemistry, University of Cambridge Lensfield Road, Cambridge CB2 1EW (UK).
Thomas O
TO
Mason
Department of Chemistry, University of Cambridge Lensfield Road, Cambridge CB2 1EW (UK).
Ulyana
U
Shimanovich
Department of Chemistry, University of Cambridge Lensfield Road, Cambridge CB2 1EW (UK).
Tuomas P J
TP
Knowles
School of Applied Sciences, RMIT University La Trobe Street, Melbourne, Victoria 3000 (Australia).
Andrew T
AT
Smith
National Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences 15 Datun Road, Chaoyang District, Beijing 100101 (China).
Elizabeth B
EB
Sawyer
National Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences 15 Datun Road, Chaoyang District, Beijing 100101 (China) ; Department of Chemistry, University of Cambridge Lensfield Road, Cambridge CB2 1EW (UK).
Sarah
S
Perrett
enzyme catalysis; immobilization; kinetics; microreactors; nanostructures