Funder identifier: FP7-KBBE-2008-2B (Other contract id)
Acronym: MAMBA
Period: July 2009 till 2013
Status: Completed
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| Institutes (8) |
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- Bangor University, more, co-ordinator
- Italian National Research Council; Institute for Coastal Marine Environment (IAMC), more, partner
- Observatoire Océanologique de Banyuls-Sur-Mer (OOB), more, partner
- Université de Bretagne Occidentale (UBO), more, partner
- Max Planck Institute for Marine Microbiology (MPIMM), more, partner
- Spanish Council for Scientific Research (CSIC), more, partner
- Heinrich-Heine University; Institute of Molecular Enzyme Technology (IMET), more
- The National Center for Scientific Research (CNRS), more
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| Abstract |
The Project aims at the mining of enzymes and metabolic pathways from extremophilic marine organisms and metagenomes from microbial communities from peculiar marine environments and consequent funnelling the new enzymatic reactions and processes towards new biotechnological applications. The project will build up on the scientific and technological excellence of individual academic and industrial partners, and beyond that, on application of the state-of-the-art technologies for archiving, molecular activity screening, protein structure elucidation, enzyme engineering and directed evolution and establishing new biotechnological processes including biocatalysis for the synthesis of fine chemicals.
Marine sampling hotspots to produce the metagenomic resources for their further exploration, and microorganisms with recently sequenced genomes will cover the whole diversity of marine microbial life at its limits (hypersaline, low and high temperature, high pressure and low water activity conditions, etc.), which is of great interest to explore the new mechanisms determining the limits to life in the biosphere, but is also of significant relevance for discovery of new industrially relevant enzymatic processes.
The central approach of the enzyme discovery will be based on the “Activity First” principle, allowing to directly access metabolic reactions of biotechnological relevance. This approach contrasts with the massive sequencing and consequent “genome/metagenome gazing” approach which can only reveal those genes encoding the enzymes of already known protein families
. A unique, hi-end functional small-molecule (SM) fluorescence screening will be employed for scoring the libraries for the activities along with the traditional chromogenic substrate assays. Individual enzymes interacting with the substrates will be identified, and in case they are new, hyperexpressed, crystallized, and their structures will be elucidated. Consequently, the most promising candidates will be scored against the batteries of chiral and non-activated substrates of relevance for biocatalysis and their ability to perform in water-free systems will be evaluated.
Designed and/or directed evolution methods will be conducted to improve the performance and specificity of the enzymes. A comprehensive bioinformatic survey throughout the whole tree of cellular life will reveal and suggest new candidates homologous to the discovered new proteins, from other organisms to be cloned and assayed. The implementation of the set of new enzymes in the biotechnological processes will be conducted in a strong alliance with competent industrial partners.
Not only we expect a quantum increase in the performance and specificity of the new biotechnological processes compared to the existing ones, but also to establish a new platform encompassing the whole set of technologies including enzyme discovery, bioinformatics, and enzyme engineering to further explore the yet undiscovered marine protein diversity space. |
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