Environmental screening for novel enzyme activities

A rapidly growing research focus involves discovery of novel enzyme activities using high-throughput functional metagenomics. In this approach, we utilize micro-expression of genes from environmental DNA libraries to identify new enzymes based on their function instead of their sequence similarity to proteins of known function. A major focus involves finding new enzymes that act upon glycans and glycoproteins. Working with collaborators at NEB and in academia, we also screen for new enzymes that manipulate DNA, RNA and proteins.

 

The first 3-D structure of a member of the GH156 family of exo-sialidases. We discovered this enzyme family using functional metagenomic screening of a fosmid library containing environmental DNA from a thermal spring. GH156 family enzymes perform catalysis via an inverting mechanism, a novel exo-sialidase mode of action.

 

References:

Zatopek, K.M., Fossa, S., Bilotti, K., Caffrey, P.J., Chuzel, L., Gehring, A.M., Lohman, G.J.S., Taron, C.H. and Gardner, A.F. (2021) Capillary electrophoresis-based functional genomics screening to discover novel archaeal DNA modifying enzymes. Appl. Environ. Microbiol. PMID: 34788065

Chuzel, L., Fossa, S.L., Boisvert, M.L., Cajic, S., Hennig, R., Ganatra, M.B., Reichl, U., Rapp, E. and Taron, C.H. (2021) Combining functional metagenomics and glycoanalytics to identify enzymes that improve structural characterization of sulfated N-glycans. Microb. Cell Fact. PMID: 34419057

Zaramela, L.S., Martino, C., Alisson-Silva, F., Rees, S., Diaz, S.L., Chuzel, L., Ganatra, M.B, Taron, C.H., Secrest, P., Zuñiga, C., Huang, J., Siegel, D., Chang, G., Varki, A. and Zengler, K. (2019) Gut bacteria responding to dietary change encode sialidases that exhibit preference for red meat-associated carbohydrates. Nat. Microbiol. PMID: 31548686

Bule, B., Chuzel, L., Blagova, E., Wu, L., Grey, M.A., Henrissat, B., Rapp, E., Bertozzi, C.R., Taron, C.H. and Davies, G. (2019) Inverting family GH156 sialidases define a new catalytic motif for glycosidase action. Nat. Commun. PMID: 31645552

Chuzel, L., Ganatra, M.B., Rapp, E., Henrissat, B. and Taron, C.H. (2018) Functional metagenomics identifies an exosialidase with an inverting catalytic mechanism that defines a new glycoside hydrolase family (GH156). J. Biol. Chem. PMID: 30249617