Periodic Reporting for period 2 - FORMILK (Innovative technology for the detection of enzyme activity in milk)
Reporting period: 2018-01-01 to 2019-12-31
Project is important for the society due to several reasons. First it brings new technologies to the milk industry for improving quality of milk and milk products by analyzing protease activity such as plasmin. Second it improves technology of preparation of lactose-free milk products. Third it accelerates the collaboration between academic institutions and Industry. Fourth it is focused on training of early stage and experienced researchers in various novel analytical methods through secondments to highly ranking laboratories in Europe, Canada and USA as well as through participation on well focused workshops and international schools."
Substantial progress was achieved in development acoustics biosensor based on β-casein layers for detection of the activity of proteases plasmin and trypsin. For the first time we used the electromagnetic piezoelectric acoustic sensor (EMPAS) for detection plasmin and trypsin cleavage with exceptionally high sensitivity of protease detection at pico molar level, which is best sensitivity reported so far. The acoustic sensor has been successfully validated in real milk samples. The protocol for detection plasmin in milk using thickness shear mode acoustic method (TSM) has been developed and validated in milk samples. The assay can be used for detection enzyme activity in dairy laboratories. The protocol has been compared with ELISA method and revealed similar sensitivity. However, in contrast with ELISA that determines only plasmin concentration, TSM provide information about plasmin activity. AFM topography of the array of short peptide substrate as well as casein layers allowed detection of the substrate topography following exposure to proteases such as plasmin and chymotrypsin. AFM method has been shown as a very effective tool for identification of bacteria in milk and milk products. The databases of potential end-users have been prepared. The miniature potentiostat, software and electrode holder has been constructed and validated. It has been demonstrated to the potential end-users
Five international workshops and three international summer schools have been organized by FORMILK consortium. In all events several PhD students and young researchers from 10 countries participated, which allows them to obtain new knowledge in application of novel physical methods in analysis of protease activity in a volume and at surfaces.