Bacteria represent an underexplored source of diglycosidases. Twenty-five bacterial strains from the genera Actinoplanes, Bacillus, Corynebacterium, Microbacterium, and Streptomyces were selected for their ability to grow in diglycosylated flavonoids-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), which was 3 to 4 orders of magnitude higher than the corresponding monoglycosidase activities. The diglycosidase production was confirmed in A. missouriensis by zymographic assays and NMR analysis of the released disaccharide, rutinose. The gene encoding the 6-O-α-L-rhamnosyl-β-D-glucosidase was identified in the genome sequence of A. missouriensis 431(T) (GenBank accession number BAL86042.1) and functionally expressed in Escherichia coli. The recombinant protein hydrolyzed hesperidin and hesperidin methylchalcone, but not rutin, which indicates its specificity for 7-O-rutinosylated flavonoids. The protein was classified into the glycoside hydrolase family 55 (GH55) in contrast to the known eukaryotic diglycosidases, which belong to GH1 and GH5. These findings demonstrate that organisms other than plants and filamentous fungi can contribute to an expansion of the diglycosidase toolbox.
- MeSH
- bakteriální proteiny genetika metabolismus MeSH
- beta-glukosidasa genetika metabolismus MeSH
- chalkonoidy chemie metabolismus MeSH
- disacharidy chemie metabolismus MeSH
- Escherichia coli genetika metabolismus MeSH
- exprese genu MeSH
- flavonoidy chemie metabolismus MeSH
- fylogeneze MeSH
- glykosidy chemie metabolismus MeSH
- hesperidin analogy a deriváty chemie metabolismus MeSH
- hydrolýza MeSH
- klonování DNA MeSH
- Micromonosporaceae klasifikace genetika metabolismus MeSH
- rekombinantní proteiny genetika metabolismus MeSH
- rhamnosa chemie metabolismus MeSH
- substrátová specifita MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
