The emergence of high-throughput sequencing (or Next Generation Sequencing NGS) in the mid-2000s, has generated an incredible amount of protein sequences deposited in the databases and the number of sequences will be increasing in the future. Nucleic and protein databases are therefore gold mine to discover novel enzymes. Genoscope is interested in large-scale functional genomics studies and had developed a high throughput platform for cloning genes and biochemical screenings of enzymes from prokaryote genomes and metagenomes. This platform is mainly use to explore the biocatalytic capabilities of enzyme families for synthetic chemistry and metabolic engineering. Here, we will illustrate the efficiency of this strategy by an overview of the main achievements obtained since CBSO 2016 on the exploration of aldolase family. Notably, we discovered in collaboration with Pr Marielle Lemaire group a family of pyruvate aldolases (PF03328) which catalyzes the aldol reaction involving unprotected hydroxypyruvate and an aldehyde offering access to valuable polyhydroxy-α-keto acids1 (de Berardinis et al., Green Chem, 2017). We discovered also that some dihydroxyacetone phosphate (DHAP)-dependent rhamnulose aldolases display an unprecedented versatility for ketones as electrophile substrates. DHAP was added as a nucleophile to several -hydroxylated ketones used as electrophiles. This aldol addition was stereoselective and produced branched-chain monosaccharide adducts with a tertiary alcohol moiety. These results contradict the general view that aldehydes are the only electrophile substrates for DHAP-dependent aldolases and provide a new CC bond-forming enzyme for stereoselective synthesis of tertiary alcohols2 (Laurent et al., Angew. Chem. Int , 2018). In addition, the need for of non-commercially available phosphorylated sugars led us to identify recently sugar kinases with broad substrate specificities3 (Vergne-Vaxelaire et al., Applied Microbiology and Biotechnology, 2018). [1] V. de Berardinis, C. Guérard-Hélaine, E. Darii, K. Bastard, V. Hélaine, T. Gefflaut, A. Mariage, J-L. Petit, N. Poupard, I. Sanchez-Moreno, M. Stam, M. Salanoubat and M. Lemaire. Green Chem, 2017, 19, 519 [2] V. Laurent, E. Darii, A. Aujon, M. Debacker, J-L. Petit, V. Hélaine, T. Liptaj, M. Breza, A. Mariage, L. Nauton, M. Traïkia, M. Salanoubat, M. Lemaire, C. Guérard-Hélaine*, and de Berardinis, V*. Angew. Chem. Int. Ed. 2018, 57, 1 – 6 [3] C. Vergne-Vaxelaire A. Mariage, J-L. Petit, A. Fossey-Jouenne, C. Guérard-Hélaine, E. Darii, A. Debard, S. Nepert, V. Pellouin, M. Lemaire, A. Zaparucha, M. Salanoubat, and V. de Berardinis*. Applied Microbiology and Biotechnology, 2018 (in press)