Objective
The primary objective of this proposal is to demonstrate the inactivation of the infective agents implicated in the causation of transmissible spongiform encephalopathies (TSEs) by the action of highly thermostable proteolytic enzymes. This will be measured using the most stringent assay available, namely the intra-cerebral inoculation of VM mice with protease-treated high-titre infectivity stocks obtained from a mouse-passaged strain of the most thermostable TSE variant reported BSE(301V).
Subsidiary objectives are:
- to show that effective inactivation can be achieved biologically at temperatures below the boiling point of water at atmospheric pressure - to show which classes and sources of thermostable protease are most appropriate for application with these agents
- the establishment and maintenance of a new breeding colony of VM mice as a resource for use in TSE research
- cloning and overexpression of genes encoding candidate proteases in heterologous mesophilic hosts enabling rapid large scale production and purification.
- to identify and purify candidate proteases for other industrial applications
Currently inactivation of TSE agents is an intractable problem, whether in medical or veterinary research, food handling and processing, or the pharmaceutical and cosmetic industries. A number of alternative solutions has been proposed and tested for this purpose. These range from chemical treatment, the use of elevated temperatures and pressures with wet heat, still higher extremes of dry heat, through to total incineration. Some chemical treatments have been shown to be ineffective (e.g. sodium hydroxide), others to be more effective but detrimental to equipment, machinery and the environment (e.g. sodium hypochlorite). Reproducible and validatable use of high temperatures and pressures requires specialised equipment, but may also be unreliable for inactivation. Incineration appears to be effective but also means that no recovery or reuse of raw materials or handling equipment can be achieved.
Reliable inactivation of TSE agents using highly thermostable proteases will be a major advance in several areas. For processes involving potentially infected animal materials (food, pharmaceutical and cosmetics industries) it will allow the safe recovery of non-proteinaceous components rather than their destruction. It will also permit the introduction of effective non-destructive disinfection cycles for most standard equipment used in handling and processing. In medical and veterinary applications, highly thermostable proteases will provide a research tool for the characterisation of causative agents of TSEs and allow a greater range of techniques to be used in their study, since dedicated equipment will not need to be destroyed after use.
The objectives of the project accord with several of the priority objectives identified in the programme work plan. The projects principal aim is to develop a biotechnological inactivation procedure for use in industry (theme 5:1). It will also have contributions to make in the characterisation of TSE agents (theme 1:1, theme 2:1, 2:3, 2.4).
Fields of science (EuroSciVoc)
CORDIS classifies projects with EuroSciVoc, a multilingual taxonomy of fields of science, through a semi-automatic process based on NLP techniques. See: The European Science Vocabulary.
CORDIS classifies projects with EuroSciVoc, a multilingual taxonomy of fields of science, through a semi-automatic process based on NLP techniques. See: The European Science Vocabulary.
- natural sciences chemical sciences inorganic chemistry alkali metals
- engineering and technology environmental engineering waste management waste treatment processes
- natural sciences biological sciences biochemistry biomolecules proteins enzymes
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Coordinator
SP4 0JG Salisbury - Wiltshire
United Kingdom
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