Flavonoid glycoside degradation can proceed through two alternative enzymatic pathways: one that is mediated by monoglycosidases, and the other catalyzed by a diglycosidase. -Diglycosidase performs the flavonoid deglycosylation in a single reaction. The characterized -diglycosidase activities recognize the following disaccharidic sugar moieties: -primeverose, acuminose, vicianose, and -rutinose. The present paper reviews the biochemical characteristics and potential industrial applications of microbial -diglycosidases that break down plant diglycoconjugated flavonoids.