Self-assembly of amyloid fibrils that display active enzymes

Xiao Ming Zhou, Aiman Entwistle, Hong Zhang, Antony P. Jackson, Thomas O. Mason, Ulyana Shimanovich, Tuomas P.J. Knowles, Andrew T. Smith, Elizabeth B. Sawyer, Sarah Perrett

Research output: Contribution to journalArticlepeer-review

39 Scopus citations

Abstract

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. Self-immobilizing enzymes: Fusion of a prion domain and a globular enzyme allows spontaneous assembly of fibrils to form an amyloid core decorated with active enzymes. Comparison of steady-state enzyme kinetic parameters for soluble and fibrillar forms of the fusion proteins indicates how immobilization affects substrate binding and catalytic turnover. The amyloid-immobilized enzymes can be recycled and reused in both batch mode and continuous flow.

Original languageEnglish
Pages (from-to)1961-1968
Number of pages8
JournalChemCatChem
Volume6
Issue number7
DOIs
StatePublished - Jul 2014
Externally publishedYes

Funding

FundersFunder number
National Natural Science Foundation of China31100564, 31110103914, 31070656

    Keywords

    • enzyme catalysis
    • immobilization
    • kinetics
    • microreactors
    • nanostructures

    Fingerprint

    Dive into the research topics of 'Self-assembly of amyloid fibrils that display active enzymes'. Together they form a unique fingerprint.

    Cite this