INVESTIGADORES
OYHENART jorge Anibal
artículos
Título:
Bacteria as source of diglycosidase activity: Actinoplanes missouriensis produces 6-O-!-L- rhamnosyl-"-D-glucosidase active on flavonoids
Autor/es:
NEHER, BARBARA; MAZZAFERRO, LAURA; KOTIK, MICHAEL; OYHENART, JORGE; HALADA, PETR; KREN, VLADIMIR; BRECCIA, JAVIER
Revista:
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
Editorial:
SPRINGER
Referencias:
Lugar: Berlin; Año: 2015
ISSN:
0175-7598
Resumen:
Microorganisms such as bacteria are still an underexplored source of diglycosidases. Twenty fivebacterial strainsfrom the genera Actinoplanes, Bacillus, Corynebacterium, Microbacterium and Streptomyces were selected according to their ability to grow ondiglycosylatedflavonoids-based media.The strains Actinoplanes missouriensis and Actinoplanes liguriae exhibited hesperidin deglycosylation activity (6-O-α-L-rhamnosyl-β-D-glucosidase activity, EC 3.2.1.168) in 2-5 orders of magnitude higher than monoglycosidase activities, respectively. The diglycosidase production was confirmed in A. missouriensis by zymographic assays and NMR analysis of the released disaccharide, rutinose. The gen for 6-O-α-L-rhamnosyl-β-D-glucosidase was identified in the genome sequence of the A. missouriensis431T(GenBank accession number BAL86042.1). The putative protein was previously classified into the glycoside hydrolase family 55 (GH55), in contrast to the reported eukaryotic diglycosidases, which belong to GH1, GH3 and GH5. The protein was functionally expressed in Escherichia coli. The purified recombinant protein deglycosylated hesperidin and hesperidin methylchalcone, but not rutin, indicating its specificity for 7-O-rutinosylated flavonoids. In agreement with the classification as GH55, transglycosylation activity was not detected. The finding of the A. missouriensis 6-O-α-L-rhamnosyl-β-D-glucosidase demonstrates that organisms other than plants and filamentous fungi could contribute to increase the diglycosidases toolbox.