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Engineering a bacterial toxin alpha-hemolysin with Vitamin B12: Single Molceule Catalysis in the alpha-hemolysin nanoreactor

Objective

This proposal describes a novel strategy for the embedding of vitamin B12 into a nano-sized protein reactor, in order to study the mechanism of reductive dechlorination reactions. This work will be the first study to observe catalysis in the alpha-hemolysin nanoreactor and will pave the way to the design of new biologically friendly catalysts for the degradation of harmful poly-chlorinated pollutants. In the experimental apparatus, preformed pores, (Staphyloccocal alpha-hemolysin) will be introduced into planar bilayers by adding small amounts of the pore protein to one chamber. Reactions are monitored by recording the modulation of the ionic current that flows through an individual pore at a fixed applied potential in millisecond time scale. The observation of catalysis in the engineered pores is our aim to provide new opportunities to examine mechanistic steps that might be missed in ensemble measurements by known techniques. We propose to insert super reduced vitamin B12 (cyanocobalamin) into alpha-hemolysin pore under anaerobic conditions. This will yield a functional nanoreactor, which can be considered as a biosensor, to detect very small amounts of contamination of chlorinated waste molecules in the environment, as well as serve as a platform for single molecule studies. These experiments will shed light into the proposed reaction mechanism on reductive dechlorination of poly-chlorinated ethenes at a molecular level of detail. Understanding the steps of the carcinogenic vinyl chloride; VC formation will provide the knowledge to construct environmental friendly, “cell free” catalysts to prevent the accumulation of these carcinogens in the environment.

Call for proposal

FP7-PEOPLE-2009-IIF
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Coordinator

THE CHANCELLOR, MASTERS AND SCHOLARS OF THE UNIVERSITY OF OXFORD
EU contribution
€ 173 240,80
Address
WELLINGTON SQUARE UNIVERSITY OFFICES
OX1 2JD Oxford
United Kingdom

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Region
South East (England) Berkshire, Buckinghamshire and Oxfordshire Oxfordshire
Activity type
Higher or Secondary Education Establishments
Administrative Contact
Linda Polik (Ms.)
Links
Total cost
No data