CORDIS provides links to public deliverables and publications of HORIZON projects.
Links to deliverables and publications from FP7 projects, as well as links to some specific result types such as dataset and software, are dynamically retrieved from OpenAIRE .
Deliverables
For soluble polymers membrane processes such as ultra- and nanofiltration in aqueous and organic solvents will be used for purification of polymers during synthesis as well as to recover them after using them in polymer supported synthesis. The use of soluble polymers should reduce mass transport limitation and allow better access to reactive sites. Since this will replace precipitation steps the influence of changing properties when attaching several groups with different polarities during the synthetic procedure can be overcome.
For protocols either for synthesis of polymers or for coupling linkers and carbohydrates to soluble polymers such as polyethylenglycol (PEG) a method for discontinuous filtration in a stirred cell has been developed. For this purpose a commercially available cell has been modified in order to be compatible with organic solvents such as THF or dichloromethane.
A protocol was developed for performing reproducible measurements under these conditions. PEG-derivatives with molecular weights of 5000g/mol could be recovered with high retention rates using commercially available ultra- and nanofiltration membranes.
This result is immediately available. Some minor modification for the protocol may be necessary depending on the polymer and the solvent used. As mainly commercially available materials are used potential user have easy access to the materials needed.
The important thing is the special know how involving which membrane is most suitable for a given separation problem as well as some special treatment of the membrane. Therefore it has to be decided on a case-by-case analysis to which extent the know-how may be licensed.
Some research is still needed for a scale up of the method. This is mainly due to technical problems when using larger filtration devices such as spirally wound modules or hollow fibre modules in organic solvents. Then there is the problem of sealing and potting. It would be desirable for further development to find a partner with suitable expertise.
Penicillin G acylase (PGA) is intended to be used for cleavage of phenylacetamide containing linkers. Sufficient amounts of enzyme have to be produced to investigate the enzyme´s ability to cleave different linkers. The PGA from E. coli ATCC11105 has been shown to be able to hydrolyse phenylacetamide containing linkers, although the cleavage efficiency has to be highly improved. A possible alternative to the E. coli enzyme is the PGA from A. faecalis ATCC19018. The enzymes show remarkable differences in their substrate spectra, their temperature stability, and activity in alkaline solutions. It can be assumed that both enzymes differ in their ability to cleave phenylacetamide containing linkers. The PGAs from E. coli ATCC11105 and A. faecalis were cloned into low copy plasmids. Both enzymes could be produced constitutively in E. coli strains containing these plasmids. Efficient production systems for recombinant PGAs from E. coli ATCC11105 and A. faecalis19018 have been established. The PGA from E. coli can be produced with enzyme activities that are at least equivalent to activities described in literature. The production of the A. faecalis enzyme was found to be even more effective. The recombinant PGAs can now be tested for their ability to cleave different phenylacetamide containing linkers. These expression systems offer the possibility for large scale production of PGA after optimisation of the fermentation conditions and downstream processing. As PGA is not only an useful enzyme for the objectives within this project it might be of potential interest to other users as well. Examples for the application of PGA which can be found in the literature cover the enzymatic synthesis of semisynthetic antibiotics, cleavage of racemates or application for protecting group techniques. First results on substrate mapping can be used to predict the enzymes selectivity.
In order to achieve carrier systems designed to fulfil the need of the biochemical application new polar, water-soluble or water-swellable polymers are developed. Preformed styrene based particles have been modified to include on the particle surfaces vinyl or phenolic groups. The vinyl modified particle has been used as a 'macromonomer' and has been successfully co-polymerised to produce supports with a tentacle structure. The commercially available PEGA-Resin has been used to couple various types of linkers for either chemical or enzymatic cleavage. PEGA-Resin is a unique polyamide/polyethylen glycol copolymer. This truly hydrophobic copolymer system provides a greater and more uniform expansion across a wide range of solvents than the PEG grafted polystyrene counterpart. For further improvement of accessibility to the linker various types of PEGA-Resins have prepared and tested. Where as the commercially available PEGA-Resin is available without restrictions the modified type is currently under further development. Therefore the amount of material is limited as well as the access is restricted to ensure first evaluation by the project partners. As soon as the problems have been solved this material will be made available as well - either on a simple commercial basis or giving licences or for joint projects.
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