Enzyme kinetics and inhibition– more information in a smaller format
A new prototype for fast and cheap measurements of enzyme characteristics has been developed by scientists at SciLifeLab. According to a new study, published in Lab on a Chip, enzyme properties in various conditions can now be studied in multiple parallel reactions using droplet microfluidic technology. This will allow large screenings of enzymes important for pharmaceutical and bioenergy development.
Enzymes catalyze biochemical reactions in our bodies and modulate processes required for life. In a wide range of disciplines, such as pharmaceutical and bioenergy development, it is important to study enzyme reaction rates, kinetics, and inhibition of enzyme processes during various conditions.
The research group Droplet Microfluidics in the Nanobiotechnology laboratory at the Royal Institute of Technology (KTH)/SciLifeLab is applying the droplet microfluidic technology for biological analysis. In droplet microfluidics, tiny water droplets in oil are produced in a microfluidic device at very high rates. Each droplet can be seen as the equivalent of a test tube and the technology allows for a much larger number of reaction vessels than any conventional platform.
One focus area for the droplet microfluidic group is on the development of new bioenergy where enzymes are used to processes biomass into fuel. The group is part of the international research center collaboration Novo Nordisk Foundation Center for Biosustainability, which aims at transforming the current chemical production to a more sustainable biobased industry. Another focus is to enable the study of enzymes in pharmaceutical research. Enzymes are often involved in disease processes and are therefore important targets for pharmaceuticals in the form of enzyme inhibitors.
‘We now have a proof-of-concept how enzyme kinetics and inhibition can be evaluated in an efficient and controlled way using very small volumes,‘ says Staffan Sjöström, PhD student in the Nanobiotechnology laboratory headed by Professor Helene Andersson Svahn. ‘I believe this system can be scaled up considerably enabling the study of a thousand different reaction conditions simultaneously. This technology can then be used to screen the performance of new enzymes to be used in for example the bioenergy industry.’
Using the device, many enzyme reaction conditions can be followed in parallel by labeling the samples with specific concentrations of a fluorescent dye – a “barcode”. Staffan and his colleagues have also custom-designed a “picoinjector” for this system that can add very small and precise volumes of enzyme substrate to the barcoded droplets and still preserve the barcode for multiplex measurement.
The study was funded by The Novo Nordisk Foundation Center for Biosustainability and the ProNova VINN Excellence Centre for Protein Technology.
Multiplex analysis of enzyme kinetics and inhibition by droplet microfluidics using picoinjectors
Sjöström, S.L., Jönsson, H.N. and Andersson Svahn, H.
Lab on a Chip, online 13 February 2013
The Science for Life Laboratory is a joint effort between four Swedish universities, Karolinska Institute, The Royal Institute of Technology (KTH), Stockholm University and Uppsala University. The centre combines advanced technology with a broad knowledge in translational medicine and molecular biosciences. SciLifeLab is a new national strategic investment in life science research that demands large-scale and specialized infrastructure. SciLifeLab has the goal to become one of the leading research centres in the world within the areas of Health and Environment.