Amyloid fibrils as a nanoscaffold for enzyme immobilization.
Amyloid fibrils are a misfolded state, formed by many proteins when subjected to denaturing conditions. Their constituent amino acids make them ideally suited as a readily functionalized nanoscaffold for enzyme immobilization and their strength, stability, and nanometer size are attractive features for exploitation in the creation of new bionanomaterials. We report successful functionalization of amyloid fibrils by conjugation to glucose oxidase (GOD) using glutaraldehyde. GOD retained activity upon attachment and successful cross-linking was determined using electrophoresis, centrifugation, sucrose gradient centrifugation, and TEM. The resulting functionalized enzyme scaffold was then incorporated into a model poly(vinyl alcohol) (PVOH) film, to create a new bionanomaterial. The antibacterial effect of the functionalized film was then tested on E. coli, the growth of which was inhibited, demonstrating the incorporation of GOD antibacterial activity into the PVOH film. The incorporation of the GOD-functionalized amyloid fibrils into PVOH provides an excellent 'proof of concept' model for the creation of a new bionanomaterial using a functionalized amyloid fibril scaffold.
Pilkington
Sarah M
SM
School of Biological Sciences, University of Canterbury, Christchurch, New Zealand.
Roberts
Sarah J
SJ
Meade
Susie J
SJ
Gerrard
Juliet A
JA
D000682:Amyloid / Q000737:chemistry* / Q000378:metabolism*; D004795:Enzyme Stability; D004800:Enzymes, Immobilized / Q000737:chemistry / Q000378:metabolism* / Q000494:pharmacology; D004926:Escherichia coli / Q000187:drug effects / Q000254:growth & development; D005949:Glucose Oxidase / Q000737:chemistry / Q000378:metabolism* / Q000494:pharmacology; D005976:Glutaral / Q000737:chemistry; D008567:Membranes, Artificial; D008826:Microbial Sensitivity Tests; D049329:Nanostructures / Q000737:chemistry*; D036103:Nanotechnology / Q000379:methods*; D011142:Polyvinyl Alcohol / Q000737:chemistry; D013499:Surface Properties
D000682:Amyloid; D004800:Enzymes, Immobilized; D008567:Membranes, Artificial; D011142:Polyvinyl Alcohol; D005949:Glucose Oxidase; D005976:Glutaral