Journal
CHEMCATCHEM
Volume 6, Issue 7, Pages 1961-1968Publisher
WILEY-V C H VERLAG GMBH
DOI: 10.1002/cctc.201402125
Keywords
enzyme catalysis; immobilization; kinetics; microreactors; nanostructures
Categories
Funding
- 973 Program [2006CB500703, 2006CB910903]
- NSFC [31110103914, 31070656, 31100564]
- CAS Fellowship for Young International Scientists [2101Y2SB01]
- NSFC Research Funds for Young International Scientists [31150110150, 31350110221]
- CAS Fellowship for Senior International Scientists [2010T1S11]
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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.
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