Identification of furostanol glycoside 26-O-β-glucosidase involved in steroidal saponin biosynthesis from Dioscorea esculenta

Steroidal saponins are natural surfactants with various biological activities, and the tubers of Dioscorea, known as yam, contain a furostanol glycoside protodioscin and a spirostanol glycoside dioscin, which are valuable saponins required for semi-synthetic production of pharmaceutical steroidal drugs. Steroidal saponins are biosynthesized from cholesterol via several steps of oxygenation and transglycosylation, and a beta-glucosidase is involved in the hydrolytic conversion from furostanol glycosides to spirostanol glycosides. To investigate steroidal saponin biosynthesis in Dioscorea spps, comparative transcriptome analysis of high saponin producers, D. esculenta and D. cayenensis, and a low producer, D. alata, was performed using 454 pyrosequencing. In this study, we isolated and characterized a beta-glucosidase ( DeF26G1) from D. esculenta. The DeF26G1 cDNA encodes a family 1 glucosidase, and the DeF26G1 transcript was present at high levels in D. esculenta but not detected in D. alata. The recombinant DeF26G1 protein hydrolyzed the 26-O-glycosidic bond of protodioscin to form dioscin, indicating that the DeF26G1 gene encodes furostanol glycoside 26-O-beta-glucosidase. These results suggested that DeF26G1 is involved in the conversion of furostanol saponins to spirostanol saponins, which seems to be related to biological defense response in the leaves of Dioscorea plants.