Objective
CONSTRUCTION OF STABLE, SPECIFIC FAT-DEGRADING ENZYMES WHICH MAY BE OF GREAT INTEREST AS ADDITIVES FOR DETERGENT OR AFTER IMMOBILIZATION ON SOLID SUPPORT COULD FORM AN EXCELLENT TOOL FOR THE SYNTHESIS OF LIPIDS OR PHOSPHOLIPIDS IN ENZYME REACTORS.
Most living organisms contain enzymes which selectively cleave certain bonds in fat thereby rendering them water soluble. Such enzymes may have industrial applications in the biotechnological production of water insoluble fine chemicals and as additives in washing detergents. This requires stable enzymes of a well defined (tailored) specificty.
A deeper insight into the specificity of (phospho)lipases, which degrade such related compounds as phospholipids and triglycerides, could lead to the design of enzymes with altered specificity for lipid molecules. As a starting point, the 3-dimensional model of the stable and high specific porcine pancreatic phospholipase A2 was used. Attempts were made to introduce by protein engineering changes that affect specificity, but not stability. For a rational design of these changes, more 3-dimensional structures are required. Thus a bacterial aspecific (phospho)lipase was isolated, characterized at the biochemical level and attempts were made to obtain its 3-dimensional structure.
The straphylococcal lipase was produced, purified and characterized. The enzyme can be described as a phospholipase A1 with significant lipase activity. Mutants were produced allowing for a more simplified purification and the introduction of a unique thiol group into the active site. Both mutations may increase the chance to solve its 3-dimensional structure. Several mutants of pancreatic phospholipase A2 were produced and characterized. 2 mutations are of special interest: mutant 1 has increase activity on neutral, but reduced activity on negatively charged phospholipids; mutant 2 has a changed chiral specificity in addition to elevated activity on uncharged phospholipids.
CONSTRUCTION BY GENETIC ENGINEERING OF AN ENZYME SHARING THE STABILITY PROPERTIES OF PHOSPHOLIPASE A2 WITH THE BROAD SUBSTRATE SPECIFICITY OF LIPASES.
BY GENETIC MANIPULATION A SERINE RESIDUE WILL BE INTRODUCED INTO THE ACTIVE SITE OF PANCREATIC PHOSPHOLIPASE A2 HAVING ITS OH FUNCTION AS CLOSE AS POSSIBLE TO THE ORIGINAL POSITION OF THE IMMOBILIZED H2O MOLECULE IN THE NATIVE MOLECULE.
TO BRIDGE THE GAP BETWEEN THE PANCREATIC PHOSPHOLIPASE A2 AND LIPASE, THE PHOSPHOLIPASE A2 AND LIPASE, THE PHOSPHOLIPASE A1 FROM ESCHERICHIA COLI WILL ALSO BE STUDIED IN DETAIL.
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.
- medical and health sciences medical biotechnology genetic engineering
- natural sciences biological sciences biochemistry biomolecules lipids
- natural sciences biological sciences genetics mutation
- natural sciences biological sciences biochemistry biomolecules proteins enzymes
You need to log in or register to use this function
Programme(s)
Multi-annual funding programmes that define the EU’s priorities for research and innovation.
Multi-annual funding programmes that define the EU’s priorities for research and innovation.
Topic(s)
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.
Data not available
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.
Call for proposal
Procedure for inviting applicants to submit project proposals, with the aim of receiving EU funding.
Data not available
Procedure for inviting applicants to submit project proposals, with the aim of receiving EU funding.
Funding Scheme
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.
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
UTRECHT
Netherlands
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.