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Extraction of Active Enzymes from "Hard-to-Break-Cells": Evaluation by a RCA-based Assay

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  • Alessio Ottaviani, Denmark
  • Cinzia Tesauro
  • S Fjelstrup, Denmark
  • Rikke Frøhlich Hougaard
  • Paola Fiorani, University of Rome, Tor Vergata, Italy
  • Alessandro Desideri, Department of Biology, University of Rome “Tor Vergata”, Via della Ricerca Scientifica, 00133, Interuniversity Consortium, National Institute Biostructure and Biosystem (INBB), Italy, Italy
  • Birgitta R. Knudsen
  • Yi-Ping Ho, Denmark
We present the utilization of a rolling circle amplification (RCA) based assay to investigate the extraction efficiency of active enzymes from a class of “hard-to-break” cells, yeast Saccaramyces cerevisiae. Current analyses of microorganisms, such as pathogenic bacteria, parasites or particular life stages of microorganisms (e.g. spores from bacteria or fungi) is hampered by the lack of efficient lysis protocols that preserve the activity and integrity of the cellular
content. Presented herein is a flexible scheme to screen lysis protocols for active enzyme extraction. We also report a gentle yet effective approach for extraction of active enzymes by entrapping cells in microdroplets. Combined effort of optimized extraction protocols and effective analytical approaches is
expected to generate impact in future disease diagnosis and environmental safety.
Original languageEnglish
Publication year5 Nov 2014
Number of pages4
Publication statusPublished - 5 Nov 2014
EventIEEE Sensors 2014 - Valencia Conference Centre, Valencia, Spain
Duration: 3 Nov 20145 Nov 2014


ConferenceIEEE Sensors 2014
LocationValencia Conference Centre

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