http://www.cnr.it/ontology/cnr/individuo/prodotto/ID11774

Glyphosate resistance by engineering the flavoenzyme glycine oxidase (Articolo in rivista)

Type

Articolo in rivista (Classe)
Prodotto della ricerca (Classe)

Label

Glyphosate resistance by engineering the flavoenzyme glycine oxidase (Articolo in rivista) (literal)

Anno

2009-01-01T00:00:00+01:00 (literal)

Http://www.cnr.it/ontology/cnr/pubblicazioni.owl#doi

10.1074/jbc.M109.051631 (literal)

Alternative label

Pedotti M; Rosini E; Molla G; Moschetti T; Savino C; Vallone B; Pollegioni L. (2009)
Glyphosate resistance by engineering the flavoenzyme glycine oxidase
in The Journal of biological chemistry (Print); American Society Of Biochemistry And Molecular Biology Inc. (ASBMB), Rockville (Stati Uniti d'America)
(literal)

Http://www.cnr.it/ontology/cnr/pubblicazioni.owl#autori

Pedotti M; Rosini E; Molla G; Moschetti T; Savino C; Vallone B; Pollegioni L. (literal)

Pagina inizio

36415 (literal)

Pagina fine

36423 (literal)

Http://www.cnr.it/ontology/cnr/pubblicazioni.owl#numeroVolume

284 (literal)

Rivista

?The ?Journal of biological chemistry (Rivista)

Http://www.cnr.it/ontology/cnr/pubblicazioni.owl#pagineTotali

9 (literal)

Http://www.cnr.it/ontology/cnr/pubblicazioni.owl#numeroFascicolo

52 (literal)

Note

ISI Web of Science (WOS) (literal)

Http://www.cnr.it/ontology/cnr/pubblicazioni.owl#affiliazioni

1. Univ Insubria, DBSM, I-21100 Varese, Italy 2. Univ Insubria, Politecn Milan, Ctr Interuniv Ric Biotecnol Prote Prot Factory, I-21100 Varese, Italy 3. Univ Roma La Sapienza, Dept Biochem Sci, I-00185 Rome, Italy 4. CNR, Inst Mol Biol & Pathol, I-00185 Rome, Italy (literal)

Titolo

Glyphosate resistance by engineering the flavoenzyme glycine oxidase (literal)

Abstract

Glycine oxidase from Bacillus subtilis is a homotetrameric flavoprotein of great potential biotechnological use because it catalyzes the oxidative deamination of various amines and D-isomer of amino acids to yield the corresponding alpha-keto acids, ammonia/amine, and hydrogen peroxide. Glyphosate (N-phosphonomethylglycine), a broad spectrum herbicide, is an interesting synthetic amino acid: this compound inhibits 5-enolpyruvylshikimate-3-phosphate synthase in the shikimate pathway, which is essential for the biosynthesis of aromatic amino acids in plants and certain bacteria. In recent years, transgenic crops resistant to glyphosate were mainly generated by overproducing the plant enzyme or by introducing a 5-enolpyruvylshikimate-3- phosphate synthase insensitive to this herbicide. In this work, we propose that the enzymatic oxidation of glyphosate could be an effective alternative to this important biotechnological process. To reach this goal, we used a rational design approach (together with site saturation mutagenesis) to generate a glycine oxidase variant more active on glyphosate than on the physiological substrate glycine. The glycine oxidase containing three point mutations (G51S/A54R/H244A) reaches an up to a 210-fold increase in catalytic efficiency and a 15,000-fold increase in the specificity constant (the k(cat)/K(m) ratio between glyphosate and glycine) as compared with wild-type glycine oxidase. The inspection of its three-dimensional structure shows that the alpha 2-alpha 3 loop (comprising residues 50-60 and containing two of the mutated residues) assumes a novel conformation and that the newly introduced residue Arg(54) could be the key residue in stabilizing glyphosate binding and destabilizing glycine positioning in the binding site, thus increasing efficiency on the herbicide. (literal)

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