Objective
Research objectives and content
Recent studies have demonstrated that cellulose binding domains (CBDs) play an important role in the activity of plant cell wall hydrolase against insoluble substrates. Pseudomonas enzymes consist of two distinct CBDs; a type II CBD domain and a second smaller domain. Although recent studies have shown that removal of both CBDs decreases the activity of Pseudomonas xylanase A (XYLA) against cellulose/hemicellulose complexes, it is unclear why Pseudomonas enzymes contain two distinct CBDs. The objective of this proposal is to determine the properties of the 40 residue CBD of XYLA and its role in the action of XYLA against the plant cell wall. Specifically, site-directed mutagenesis will be used to change the conserved aromatic residues to alanine to establish whether they play a pivotal role in binding cellulose, or are integral to the correct folding of the CBD. To establish the importance of the CBD in the activity of XYLA, xynA will be engineered such that it encodes two derivatives of XYLA in which either the large type II CBD (XYLA') or the small CBD (XYLA") has been removed. Analysis of the capacity of XYLA, XYLA' and XYLA" to hydrolyse cellulose/xylan complexes will establish why Pseudomonas glycosyl hydrolase, in general, have two distinct CBDs.
Training content (objective, benefit and expected impact)
This project will provide me with an excellent training in the use of recombinant DNA technology (site-directed mutagenesis, PCR, DNA sequencinge etc) to study the structure and function of proteins. I already have experience in the use of both biochemical and chemical techniques to study industrially important enzymes. At the completion of my training I will be able to use recombinant DNA technology to produce large quantities of these proteins, and engineer their structure to underpin more sophisticated studies on these important biocatalysts.
Links with industry / industrial relevance (22)
Professor Gilbert has strong industrial contacts with Zeneca BioProducts (enzymes in the Paper Pulp Industry), Thomas Swan (Paper pulp Industry), Genacor (Detergent Industry), Dalgety and FinnFeeds International (Animal Feeds and the Food industry).
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 biological sciences genetics DNA
- natural sciences chemical sciences catalysis biocatalysis
- engineering and technology industrial biotechnology biomaterials
- natural sciences biological sciences biochemistry biomolecules proteins enzymes
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Programme(s)
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Multi-annual funding programmes that define the EU’s priorities for research and innovation.
Topic(s)
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Calls for proposals are divided into topics. A topic defines a specific subject or area for which applicants can submit proposals. The description of a topic comprises its specific scope and the expected impact of the funded project.
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Procedure for inviting applicants to submit project proposals, with the aim of receiving EU funding.
Funding Scheme
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Funding scheme (or “Type of Action”) inside a programme with common features. It specifies: the scope of what is funded; the reimbursement rate; specific evaluation criteria to qualify for funding; and the use of simplified forms of costs like lump sums.
Coordinator
NE1 7RU Newcastle - Upon Tyne
United Kingdom
The total costs incurred by this organisation to participate in the project, including direct and indirect costs. This amount is a subset of the overall project budget.