Work Package 1
Development of Libraries of ω-Transaminases and HTS Method.
This Work Package, lead by UNIMAN, will provide new and improved transaminase enzymes for use in the other WP’s. High through-put screening methods to aid in directed evolution of these enzymes will be developed and documented.
New enzymes will be identified from a culture collection of wild type organisms, and also by using bioinformatics to identify putative genes from available databases. These genes will be cloned into suitable microorganisms for enzyme expression, mutation and screening for activity. This will build on expertise in these techniques already used at CoEBio3 at Manchester to develop new enzyme families.
HTS methods will be developed based upon technologies already in use by CoEBio3. Collaboration with UNIGRO will allow designer mutants to be created based on molecular engineering. Directed evolution will also allow specific improvement of enzymes to meet substrate and engineering demands (substrate specificity, stereochemistry, activity, solvent and thermal stability). Collaboration with UNIGRAZ will develop co-factor issues to be addressed in WP2. Enzyme and reactor engineering will then be addressed by DTU, CLEA and EVONIK to develop other cascades.