title: Cloning and characterization of saponin hydrolases from Aspergillus oryzae and Eupenicillium brefeldianum.
abstract: We purified saponin hydrolases from Aspergillus oryzae PF1224 and Eupenicillium brefeldianum PF1226. It was confirmed that the enzymes from A. oryzae PF1224 (Sda1) and E. brefeldianum PF1226 (Sde1) are glycoproteins with molecular masses of 82 and 90 kDa respectively. The deduced amino acid sequences of each enzyme from the cloned genes (sda1 or sde1) showed approximately 50% homology with that of the saponin hydrolase Sdn1 from Neocosmospora vasinfecta var. vasinfecta PF1225 (DDBJ accession no. AB110615). When sda1 and sde1 were expressed in the host Trichoderma viride under the control of the cellobiohydrolase I gene promoter, recombinant proteins were secreted with molecular masses of 77 and 67 kDa respectively. These recombinant enzymes hydrolyzed soyasaponin I to soyasapogenol B and triose, and its substrate specificities for glycosides were similar to that of Sdn1, but the specific activities of these enzymes were lower than that of Sdn1.
authors.affiliation: Microbiological Resources and Technology Laboratories, Meiji Seika Kaisha, Ltd, Kanagawa, Japan. manabu_watanabe@meiji.co.jp
authors.forename: Manabu
authors.initials: M
authors.lastname: Watanabe
authors.forename: Naomi
authors.initials: N
authors.lastname: Sumida
authors.forename: Koji
authors.initials: K
authors.lastname: Yanai
authors.forename: Takeshi
authors.initials: T
authors.lastname: Murakami
mesh_terms: D000595:Amino Acid Sequence; D001236:Aspergillus oryzae / Q000201:enzymology*; D003001:Cloning, Molecular; D006023:Glycoproteins; D006867:Hydrolases / Q000737:chemistry / Q000378:metabolism*; D008970:Molecular Weight; D009828:Oleanolic Acid / Q000031:analogs & derivatives / Q000378:metabolism; D012503:Saponins / Q000378:metabolism*; D017385:Sequence Homology; D013379:Substrate Specificity; D014306:Trioses / Q000378:metabolism
chemical_list: D006023:Glycoproteins; D012503:Saponins; D014306:Trioses; C407096:soyasapogenol B; C056684:soyasaponin I; D009828:Oleanolic Acid; D006867:Hydrolases