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Ultra High-throughput platform for the selection of thermostable proteins by thermophilic in vitro transcription-translation and microfluidics (HOTDROPS)
Date du début: 1 juin 2013, Date de fin: 31 mai 2017 PROJET  TERMINÉ 

"This initiative connects four companies and four academic groups in pursuit of a central biotechnological challenge: to identify and improve enzymes for application in industrial biocatalysis. The technology interchange is centered around key technologies to generate thermostabilized proteins. We will develop, share and spread the following approaches (i) folding interference assays with a thermostable fluorescent protein reporter, (ii) high throughput assays in microfluidic droplets that allow screening of libraries on the order of 10e8, (iii) in vitro expression in microdroplets using expression extracts from themophiles, and (iv) approaches to generate enzyme libraries from metagenomic sources and by randomization of existing proteins. Suitable assays will be established for a list of target enzymes of interest to the industrial partners (most prominently hydrolases – such as esterases, cellulases and proteases), ensuring wide applicability and industrial relevance. Improved mutants will be characterized biochemically and scale-up and optimization for protein production by fermentation techniques will be attempted. An experienced partner who will enable others by workshop training, interchange of technical and academic personnel and joint projects represents each of these approaches. The objective of this application is to make the considerable technical skill involved in molecular biology, enzymology and screening technologies available to a wider circle of users, ranging from academic labs to very recent spin-off companies and the world’s largest provider of industrial enzymes."

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