1 / 16

Prion Degradation by a Feather-Degrading Keratinase

Prion Degradation by a Feather-Degrading Keratinase. Jason Shih North Carolina State University. Keratinase Technology. Purified enzyme, MW = 33 kDa Gene isolated and sequenced Genetic modification for over-expression Scale-up production by fermentation Feather process Feed enzyme

giles
Télécharger la présentation

Prion Degradation by a Feather-Degrading Keratinase

An Image/Link below is provided (as is) to download presentation Download Policy: Content on the Website is provided to you AS IS for your information and personal use and may not be sold / licensed / shared on other websites without getting consent from its author. Content is provided to you AS IS for your information and personal use only. Download presentation by click this link. While downloading, if for some reason you are not able to download a presentation, the publisher may have deleted the file from their server. During download, if you can't get a presentation, the file might be deleted by the publisher.

E N D

Presentation Transcript

  1. Prion Degradation by a Feather-Degrading Keratinase Jason Shih North Carolina State University

  2. Keratinase Technology • Purified enzyme, MW = 33 kDa • Gene isolated and sequenced • Genetic modification for over-expression • Scale-up production by fermentation • Feather process • Feed enzyme • Prion degradation

  3. Transmissible Spongiform Encephalopathy (TSE): A Timely and Global Issue • Over 180,000 cases of Mad Cow Disease (or Bovine Spongiform Encephalopathy, BSE) have been reported in Europe. • Correlated with cases of Creutzfeldt-Jakob Disease (CJD), a fatal neurodegenerative disease in humans. • No effective diagnostics, vaccines or therapeutics are available for BSE or CJD. • Concerns about BSE and CJD transmission have forced a re-examination of animal feed production and hospital safety measures, respectively.

  4. Prion Structure PrPC PrPSc

  5. keratinase Degradation Treating Prion Proteins with keratinase Conformational Change PrPSc Infectious, insoluble PrPC Non-infectious, soluble

  6. “Proof of Concept” BSE Experiment 1 2 3 4 5 6 7 8 9 10

  7. Brain Stem Tissue Homogenization Heat pre-treatment (115o C - 40 min) Keratinase digestion (50o C - 1hr) MeOH precipitation Redissolved in buffer SDS-PAGE Western blotting and detection

  8. Pre-heat and Keratinase on tissuewhole homogenates and supernatants

  9. Effects ofN-lauroylsarcosine (sarc) and KE

  10. Detergent Effect

  11. Enzymatic degradation of prionsfrom different individual animals

  12. Enzyme Concentrations

  13. Digestion Time

  14. Temperature Effect

  15. Digestion by various enzymes

More Related