Identification and cloning of the first O-demethylase gene of isoflavones from Bifidobacterium breve INIA P734

Biochanin A and formononetin are methylated isoflavones, abundant in legumes, which can be converted genistein daidzein by means of an O-demethylation reaction. Our objective was to transform these isoflavones into higher biological activity bioavailability compounds, such as genistein. Prior screening, using 10 lactic acid bacteria (LAB) 3 bifidobacteria strains, revealed that only 7 lactobacilli strains were able biochanin genistein, while none the demethylated formononetin. As dmt734 gene from Bifidobacterium breve INIA P734 putatively annotated O-demethylase, it cloned a vector expressing antibiotic resistance (pNZ:TuR.dmt734) well food grade (pLEB590.dmt734), subsequently transformed LAB O-demethylase lacking strains. Those recombinant harbouring pNZ:TuR.dmt734, or pLEB590.dmt734, managed but did not daidzein. These results confirm demethylation specificity biotechnological interest B. activity, would value for development fermented vegetable-based foods enriched