DegradePlant · Directed evolution and semi-rational engineering of plant cell wall-degrading enzymes
FP7 — People (Marie Curie Actions)
- Duration
- 2011-06-01 → 2013-05-31
- EU contribution
- €200,050
- Participants
- 1
- Scheme
- MC-IEF
Lines connect the coordinator with its partners.
Results in brief
Directed evolution and semi-rational engineering of plant cell wall-degrading enzymes
The aim of this project is to investigate the role of insertions and deletions (indels) in the acquisition of new enzyme functions. More specifically, our aim is to perform laboratory-evolution experiments incorporating indels to investigate whether backbone modifications can be selected for functional effects and how these changes constrain further evolutionary optimisation. Given the lack of readily facile techniques to generate libraries of indel variants, a major aspect of this project was focused on developing new methodologies and tools for random indel mutagenesis. Besides the use of the initial models of choice (i.e. glycoside hydrolases), we also expanded this project towards the use of other (promiscuous) enzymes, including a phosphotriesterase from P. diminuta (PTE) and an arylsulfatase from P. aeruginosa (PAS).
Data: CORDIS, © European Union
Project objective
This proposal focuses on the development of new techniques and strategies for the directed evolution of enzymes useful for industrial (white) biotechnology, including the development of new genetic methodologies to improve the potential of variant libraries, as well as new screening tools to broaden the range of targets for directed evolution. Specifically, the proposed project will apply these new methodologies to plant cell-wall degrading enzymes, namely cellulases and hemicellulases, which are crucial biocatalysts for the production of second-generation lignocellulose-derived biofuels. The enzymes targeted in this proposal are phylogenetically-related, share the same fold, catalyze the same chemical transformation on a large number of structurally related substrates and thus provide good starting points to switch substrate specificities by directed evolution. In order to achieve these challenging objectives, the proposed project will combine new technical advances: on the one hand methodologies to construct gene libraries and stability compensation strategies, and on the other hand new screening approaches of unprecedented throughput based on compartmentalized screening in microdroplets that are processed in microfluidic devices.
Original text from CORDIS.
Participants
- THE CHANCELLOR MASTERS AND SCHOLARS OF THE UNIVERSITY OF CAMBRIDGE · CAMBRIDGECoordinatorUnited Kingdom
Links
Data: CORDIS, © European Union