title: Production of steviol from steviol glucosides using β-glycosidase from Sulfolobus solfataricus.
abstract: Steviol is a diterpene isolated from the plant Stevia rebaudiana that has a potential role as an antihyperglycemic agent by stimulating insulin secretion from pancreatic beta cells and also has significant potential to diminish the renal clearance of anionic drugs and their metabolites. In this study, the lacS gene, which encodes a thermostable β-glycosidase (SSbgly) enzyme from the extremely thermoacidophillic archaeon Sulfolobus solfataricus, was cloned and expressed in E. coli Rossetta BL21(DE3)pLyS using lactose as an inducer. Through fermentation, SSbgly was expressed as a 61kDa protein with activity of 24.3U/mg and the OD600of 23 was reached after 18h induction with 10mM lactose. Purified protein was obtained by Ni-Sepharose chromatography with a yield of 92.3%. SSbgly hydrolyzed steviol glycosides to produce steviol with a yield of 99.2%. The optimum conditions for steviol production were 50U/ml SSbgly and 90mg/ml Ste at 75°C as determined by the response surface method.
authors.affiliation: Institute of Food Industrialization, Institutes of Green Bio Science & Technology, Seoul National University, Pyeongchang-gun, Gangwon-do, 25354, South Korea.
authors.forename: Thi Thanh Hanh
authors.initials: TT
authors.lastname: Nguyen
authors.affiliation: CJ CheilJedang, Life Ingredient & Material Research Institute, Suwon, 16495, South Korea.
authors.forename: Seong-Bo
authors.initials: SB
authors.lastname: Kim
authors.affiliation: Section of Neurobiology, Department of Biological Sciences, University of Southern California, Los Angeles, CA, 90089, USA.
authors.forename: Nahyun M
authors.initials: NM
authors.lastname: Kim
authors.affiliation: OTTOGI Corporation, Anyang, Kyunggi, 06177, South Korea.
authors.forename: Choongil
authors.initials: C
authors.lastname: Kang
authors.affiliation: OTTOGI Corporation, Anyang, Kyunggi, 06177, South Korea.
authors.forename: Byoungsang
authors.initials: B
authors.lastname: Chung
authors.affiliation: Skin Research Institute, Amorepacific Corporation R&D Center, Yongin, 17074, South Korea.
authors.forename: Jun-Seong
authors.initials: JS
authors.lastname: Park
authors.affiliation: Institute of Food Industrialization, Institutes of Green Bio Science & Technology, Seoul National University, Pyeongchang-gun, Gangwon-do, 25354, South Korea; Graduate School of International Agricultural Technology, Seoul National University, Pyeongchang-gun, Gangwon-do, 25354, South Korea. Electronic address: kimdm@snu.ac.kr.
authors.forename: Doman
authors.initials: D
authors.lastname: Kim
mesh_terms: D019843:Archaeal Proteins / Q000378:metabolism*; D045786:Diterpenes, Kaurane / Q000096:biosynthesis* / Q000737:chemistry / Q000378:metabolism; D004795:Enzyme Stability; D005959:Glucosidases / Q000235:genetics / Q000378:metabolism*; D005960:Glucosides / Q000737:chemistry / Q000378:metabolism; D006358:Hot Temperature; D007004:Hypoglycemic Agents / Q000378:metabolism; D007700:Kinetics; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D048229:Sulfolobus solfataricus / Q000201:enzymology* / Q000235:genetics; D013549:Sweetening Agents / Q000378:metabolism
chemical_list: D019843:Archaeal Proteins; D045786:Diterpenes, Kaurane; D005960:Glucosides; D007004:Hypoglycemic Agents; D011994:Recombinant Proteins; D013549:Sweetening Agents; C012043:stevioside; C012042:steviol; D005959:Glucosidases; C104851:Sulfolobus solfataricus beta-glycosidase
keywords: Optimization; Steviol; Stevioside; Sulfolobus solfataricus; β-glycosidase