[14-ERA IB] MetaCat: A metagenomic collection of novel and highly efficient biocatalysts for industrial biotechnology

Chemical and bio-industries have a steadily growing demand for enzyme biocatalysts, which can catalyze a huge variety of different chemical reactions with high activity, substrate specificity and enantioselectivity. However, enzymes have been evolved by nature to work in living cells and under mild reaction conditions; consequently, most enzymes cannot be applied in industrial processes directly. Although it is known that few enzymes exist, which are well suited for biotechnological applications, the molecular basis is unknown. The MetaCat project will deliver innovative tools
and knowledge for the identification of such robust "all-round frequent hit" enzymes (AFHs).
Metagenomic resources will be exploited by novel function- and sequence-based screenings to identify nitrilases, transaminases, ketoreductases, glycosyl hydrolases and lipases/esterases. A combination of metagenomics and metacatalysis will be used together with structure-based and
high-throughput technologies, generic model substrates mimicking challenging chemical synthesis steps, next generation sequencing technologies as well as in silico data mining. New genetic tools using synthetic biology approaches will be developed to construct a cell-free function-based
screening platform for faster and improved screening. An innovative single-cell laser trapping technology will be established to give access to a new previously unknown enzyme diversity. The identified enzymes will form a marketable versatile biocatalyst collection together with a comprehensive database and can be used as starting points for modeling and in vitro evolution experiments. They will be applied to improve chemical production processes regarding e.g. timelines, purity of the products, environmental sustainability and will lead to value-added products.
Thus, MetaCat will contribute to shorten timelines for development of biotechnological processes and thus to make industrial biocatalysis more attractive and profitable.

Main Project Objectives
-Functional and highly diverse collections of most wanted AFHs serving as novel and improved biocatalysts for direct application in industrial processes.
-An innovation pool for the detection of enzymes: innovative cell-free and single-cell tools for functional metagenome and genome mining.
-Biochemical and bioinformatic tools to identify AFH enzymes.
-Understanding of structures and mechanisms underlying enzyme promiscuity.
-Optimized and efficient chemical production processes
-High value-added products, i. e. platform and fine chemicals and pure enantiomers.

The objectives will be achieved by the following Work Packages
WP 1. Functional metagenomics and screenings (Golyshin)
WP 2. NGS metagenome analyses, sequence-based enzyme identification (Ferrer)
WP 3. Data integration, advanced bioinformatics and software development (Vallenet/Zaparucha)
WP 4. Novel and advanced screening hosts and cell-free metagenomics (Streit/Chow)
WP 5. Enzyme expression, biochemical and structural characterization (Jaeger)
WP 6. Biocatalytic process implementation (Koch)
WP 7. Project management and coordination (Streit/Chow)

The Bangor University-lead WP 1 is broken down in following tasks:
T1. Sample collection
Our resources comprise a wide range of terrestrial, marine, plant- and animal-derived samples containing all forms of microbial life and extremely high diversity. The consortium has access to a wide range of biodiversity that is unique and can in part not be sampled by other groups or companies.
T2. Enrichments with industry-relevant substrates
Enrichments will be set up with samples from some of the sites. The samples will be enriched with model substrates provided by industry partners. Further, aerobic but also micro-aerobic and anaerobic conditions for the enrichments will be established. Especially the anaerobic technologies will allow unraveling novel communities and microbes that have not been addressed so far.

Potential of the expected results
The potential for industrial applications of the MetaCat enzyme collection is extremely high as they enable efficient production of high-value chemical compounds. The MetaCat participating companies will promote the newly established screening methods and will distribute knowledge and understanding about structural features of enzymes. They put strong emphasis on direct implementation of the identified biocatalysts in industrial production processes and evaluate the whole approach at an early stage, e.g. concerning challenging synthesis steps. Further, the novel technologies developed (esp. the cell-free systems) will ultimately result in a strong leadership.

Environmental benefit
Enzyme application significantly lowers the use of often highly toxic conventional catalysts and reduces environmental burden. Biocatalytic processes decrease energy consumption and costs as they can usually be carried out at lower temperatures and with fewer reaction steps, i.e. milder process conditions. Thus, the use of organic solvents can be reduced; unwanted by- and waste-products can be circumvented or converted. Renewable resources can be used more efficiently leading to a higher sustainability. A bio-efficiency study can be commissioned to evaluate benefits of novel enzymatic reactions and improved or novel industrial processes.

Integration of the whole value-added chain
The MetaCat consortium members cover the whole value chain. Novozymes is world leader in finding bio-industrial solutions and evocatal is expert in the production of enzymes and fine chemicals. Bayer is one of the most important chemical and pharmaceutical companies and will distribute the enzymatically produced value-added products worldwide. All companies are highly interested to implement and further commercialize the developed screenings and the resulting enzymes.