Moore, B.D. and Stevenson, L. and Watt, A. and Flitsch, S. and Turner, N.J. and Cassidy, C. and Graham, D. (2004) Rapid and ultra-sensitive determination of enzyme activities using surface-enhanced resonance Raman scattering. Nature Biotechnology, 22 (9). pp. 1133-1138. ISSN 1087-0156Full text not available in this repository. (Request a copy from the Strathclyde author)
Measurement of enzyme activity and selectivity at in vivo concentrations is highly desirable in a range of fields including diagnostics, functional proteomics and directed evolution. Here we demonstrate how surface-enhanced resonance Raman scattering (SERRS), measured using silver nanoparticles, can be used to detect the activity of hydrolases at ultra-low levels. This approach was made possible by designing 'masked' enzyme substrates that are initially completely undetected by SERRS. Turnover of the substrate by the enzyme leads to the release of a surface targeting dye, and intense SERRS signals proportional to enzyme activity are generated. The method was used to rapidly screen the relative activities and enantioselectivities of fourteen enzymes including examples of lipases, esterases and proteases. In the current format the sensitivity of the technique is sufficient to detect 500 enzyme molecules, which offers the potential to detect multiple enzyme activities simultaneously and at levels found within single cells.
|Keywords:||enantioselective hydrolases, ph indicators, chemistry, catalysts, silver, serrs, benzotriazole, spectroscopy, resolution, libraries, Chemistry, Biomedical Engineering, Applied Microbiology and Biotechnology, Bioengineering, Molecular Medicine, Biotechnology|
|Subjects:||Science > Chemistry|
|Department:||Faculty of Science > Pure and Applied Chemistry
|Depositing user:||Mr Derek Boyle|
|Date Deposited:||19 May 2006|
|Last modified:||29 Apr 2016 07:41|