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 .
Exploitable results
Stabilizer screening for penicillin acylase (PA) was carried out in order to increase the stability of the enzyme with time at high temperatures. The polyelectrolyte screen revealed the positively charged DEAE-dextran (0.25%) stabilized the PA by a factor of 10. The examination of covalently coupling PA to dextran did not result in a vast improvement in the stability of PA over that combined with DEAE-dextran. The addition of high concentrations of lactitol (50% w/v) working in combination with the polyelectrolyte led to an increase in 3/2 life from 12 minutes in the presence of DEAE-dextran alone to 20 hours in combination at 60degreesC.
Isoelectrofocusing electrophoresis can be used as a method to measure the ratio enzyme-polyelectrolyte: this information should allow to modulate respective concentrations to reach best conditions.
Investigation of the stoichiometry of binding between DEAE-dextran and PA by gel electrophoresis revealed a molar ratio of enzyme to polyelectrolyte of 10:1. This can be utilized as a measure of the efficiency of polymer binding. Decreased levels of polyelectrolyte can be used to stabilize the enzyme in the final process, thus reducing cost and adverse effects to the biochemical processes.
The screening of stability conferred to trypsin in the presence of polyelectrolytes is being carried out. Buffer and pH screening reveals trypsin is more stable between pH 6 and 7 in Sodium phosphate or Bistrispropane, at 100mM. Protein/electrolyte interactions are not detectable using IEF due the very basic nature of the protein. Native agarose gel electrophoresis appears to have overcome these difficulties.
entities potentially involved. A new smart polymer was produced and characterized: a copolymer of two compounds with solubility properties that can be finely tuned by varying the proportion between the two monomers. In this way it can be obtained a polymer soluble up to a defined temperature at a certain pH, or vice versa, and both temperature and pH can be decided in agreement with enzyme need, and not approximate best enzyme conditions to factors already defined by polyelectrolyte characteristics. This is of great importance when it necessary to work with very expensive enzymes, or to improve processes that are already working at a good level.
Results of this research can be useful to all chemical industries, mainly those involved reactions using biocatalysts.
Trypsin and aminopeptidase immobilized on this commercial preparation show improved stability, can be easily recovered and maintain activity after recovery.
Trypsin is used to obtain polylysines: the highest degrees of polymerisation were obtained at 100 % conversion which is of greatest interest. Furthermore no strong interactions between the catalyst and the products were noticed: the recovery of the catalyst is thus very easy to proceed. This process is very interesting as it gives a lot of information on technical and economic aspects: it is possible to compare several different reaction conditions (not only immobilized-free enzyme) and find the minimum cost for a pure enzyme where it is favourable to use immobilization techniques.
Results of immobilized aminopeptidase are encouraging, as the cost of preparation of this enzyme are very high, making very important the opportunity to reuse it in several synthesis processes. Some problems have been found so far due to the poor purification level of the enzyme preparation, so this process will be very useful for the evaluation of improvement of results (both regards to production and scientifically) versus industrial costs of the whole process.
Results of this research can be useful to all chemical industries, mainly those involved reactions using biocatalysts.
Searching for OpenAIRE data...
There was an error trying to search data from OpenAIRE
No results available