Biomass hydrolyzing enzymes identified by functional screening of a metagenomic library from algal biofilms.

Biomass hydrolyzing enzymes are increasingly searched for the production of biofuels and renewable chemical compounds using biomass. Microorganisms living on algaes are an interesting reservoir of biomass hydrolyzing enzymes, as they are in constant interaction with algal biomass. Therefore, we are interested in looking for some of those enzymes synthesized by the microflora living on the surface of the brown algae Ascophyllum nodosum. Algae samples were collected in the winter 2012 and a microbial DNA extraction method was developed. The whole extracted microbial genomes of the microorganisms living on the algae were restricted, inserted in a cloning vector and ligated products were used for transformation of cultivable Escherichia coli host cells. This metagenomic library was then screened for diverse enzymatic activities (lipolytic enzymes, cellulases, beta-glucosidases, alpha-amylases, arabinanases, xylanases and proteases) on agar plates with specific substrates. Five putative lipolytic enzymes, one cellulase and one beta-glucosidase were identified. Sequence analysis revealed low (<50%) sequence identities with known enzymes sequences, meaning new enzymes from unknown genomes have been discovered. To our knowledge this is the first functional screening that was realized with a metagenomic library from algal biofilms and this is the first cellulase identified by marine metagenomics. A second library has been constructed from algae sample from summer 2012 and is currently being screened. New enzymatic tests are being developed for the identification of enzymes degrading specific algal polysaccharides like agarases, carrageenases, alginate lyases, laminarinases,… Those very specific enzymes aren’t well known yet, and our metagenomic approach will probably help us to identify new families and structures of those algal biomass hydrolyzing enzymes.