The eurobiosyn project is structured into the following workpackages.
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The prime objective of this Work Package is the development of a switch for directed re-programming of the bacterial proteome. This will be achieved through (1) the development of a general procedure for in silico design and engineering target sites for viral proteases within permissive loops of given bacterial target proteins, (2) Insertion of multiple SceI-sites through the chromosome of a recipient target strain and (3) engineering of a phage lambda-based genetic switch able to simultaneously enhance expression of desired enzyme, inactivate target proteins/activities, and trigger chromosome destruction.
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The goal of this Work Package is the overall SBT-design and application, comprising (1) the identification and qualification of host genes that potentially interfere with monosaccharide production and (2) SBT production and application.
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Once the first SBT is isolated from the remaining metabolic network, it will still obey the enzymatic constraints of cell homeostasis. These constraints will become especially evident for product accumulation which most likely would be very difficult with a SBT having only built-in the “natural” set of enzymes. To improve the SBT’s efficiency, enzymes responsible for constraints in productivity have to be identified and to be reengineered in order to display properties, which lead to higher SBT productivity and also allow product accumulation.
The identification of productivity constraints in the isolated SBT will be achieved through a detailed system analysis of a SBT using highly dynamically resolved experimental data, stochastic grey-box modelling and optimal experimental design tools as well as mathematical optimisation of SBT productivity.
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Bottlenecks in the SBT originating from enzymes with insufficient stability, activity, or inhibition by substrate or product will be removed by protein engineering of these enzymes or by replacing them by homologues with desired properties.
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Project Management has the objective to coordinate the overall technological progress, administration and finances of the project, to communicate with the EU commission and to coordinate and supervise the multidisciplinary team of four European Partners to realise deliverables and milestones according to the Work Plan.
The second part of objectives is to establish a framework for the efficient dissemination of the results of EUROBIOSYN, to generate wide visibility for the Project and its results and to establish the conditions and network for a larger European SYNTHETIC BIOLOGY initiative that should result in the application for a NoE or an IP.
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