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Monitoring of total antioxidant activity in different beverages using biosensors technology

Objectif

Recently, it was shown that the antioxidants have large and essential influence on human health. It is assumed that the antioxidants are protective against cardiovascular disease and certain form of cancer, and perhaps against other diseases also. These protective effects have been attributed, in large part, to the antioxidants derived from food products including antioxidant nutrients vitamin C and b-carotene, and polyphenol flavonoids. The polyphenolic flavonoids, which showed antioxidant activity, differ from each other in number and arrangement of the hydroxyl groups, as well as from the nature and extent of alkylation and/or glycosylation of these groups. Thus, their content can be estimated by polyphenol oxidases, such as laccases and tyrosinases. Most of their dietary flavonoids derived from beverages (tea, coffee, juices, wine and others), fruits and vegetables. Taking into account that beverages are one of the most consumable food products we developed the method of the determination the total antioxidant activity in beverages based on polyphenol oxidases and biosensor technology through the determination of total polyphenolic flavonoids content. Therefore, the proposed assay can allow the identification of beverages and food products that offer potentiale antioxidant protection. Moreover, this method may help to identify the antioxidants and may be used through investigation of their bioavailability, absorption and metabolism as well as their pharmacokinetic properties.
The main ain of the proposed research project is to develop novel and efficient on line biosensor based on phenol oxidases for the determination of the total antioxidant activity of beverages. We propose a new approach to rational design of the biosensor combined fundamental and all pied investigations. The fundumental study (biochemical, physico-chemical and catalytic properties, and mechanism of action) of enzymes, involved in the biosensor as the biological component will be used in developing of biosensor. The most important characteristics of the proposed biosensor will be simplicity in use, rapidity of assay and low cost of analysis. Thus, the development of such biosensor besides its economic impact would also assist to further progress in the field of food quality assessment. The research subject and the objectives completely conform to the scope of the INCO-Copernicus call. In order to accomplished the proposed research the scientific teams with different experience are involved in the project. The efficiency and quality of the biosensor developed within the project will be industrially tested on the beverages produced by the Ukrainian small enterprise, participating in the project. This would be the first step for implementation of the proposed system on the industrial scale and would benefit both EU and NIS countries.

Our specific tasks within the project are the following:
a) Characterization and investigation of the mechanism of action and stabilization of laccases and tyrosinase. The biochemical and physico-chemical properties, substrate specificity, stability, mechanism of action (including mechanism of bioelectrocatalysis) of laccases and tyrosinases will be studied. Different approaches for enzyme long-term function will be developed;
b) Development of novel methods for immobilization of polyphenol oxidases as a key constituting part of the biosensor. Construction of a prototype flow-injection biosensor with electrochemical detection for the determination of total antioxidant activity of beverages. The operational stability, the assay time and sensitivity of proposed biosensor will be studied;
c) Comparative study of alternative methods for the determination of total antioxidant activity in beverages. Different approaches to eliminate the influence of different compounds in beverages on precision and accuracy of the measurement will be developed;
d) Industrial application of the proposed biosensor.

Wide range of the microbiological, biochemical and physico-chemical techniques will be used within this project. Among the most important are: fermentation, liquid and gas-phase chromatography, electrophoresis, isoelectrofocusing, polarographic, amperometric and electrochemical methods, NMR, NMRD, EPR, UV-visible and CD-spectroscopies.

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Coordinateur

University of Bologna
Contribution de l’UE
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Adresse
Viale Berti Pichat, 10
40127 Bologna
Italie

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