title: UV-dependent production of 25-hydroxyvitamin D2 in the recombinant yeast cells expressing human CYP2R1.
abstract: CYP2R1 is known to be a physiologically important vitamin D 25-hydroxylase. We have successfully expressed human CYP2R1 in Saccharomyces cerevisiae to reveal its enzymatic properties. In this study, we examined production of 25-hydroxylated vitamin D using whole recombinant yeast cells that expressed CYP2R1. When vitamin D3 or vitamin D2 was added to the cell suspension of CYP2R1-expressing yeast cells in a buffer containing glucose and β-cyclodextrin, the vitamins were converted into their 25-hydroxylated products. Next, we irradiated the cell suspension with UVB and incubated at 37 °C. Surprisingly, the 25-hydroxy vitamin D2 was produced without additional vitamin D2. Endogenous ergosterol was likely converted into vitamin D2 by UV irradiation and thermal isomerization, and then the resulting vitamin D2 was converted to 25-hydroxyvitamin D2 by CYP2R1. This novel method for producing 25-hydroxyvitamin D2 without a substrate could be useful for practical purposes.
authors.affiliation: Department of Biotechnology, Faculty of Engineering, Toyama Prefectural University, 5180 Kurokawa, Imizu, Toyama 939-0398, Japan.
authors.forename: Kaori
authors.initials: K
authors.lastname: Yasuda
authors.forename: Mariko
authors.initials: M
authors.lastname: Endo
authors.forename: Shinichi
authors.initials: S
authors.lastname: Ikushiro
authors.forename: Masaki
authors.initials: M
authors.lastname: Kamakura
authors.forename: Miho
authors.initials: M
authors.lastname: Ohta
authors.forename: Toshiyuki
authors.initials: T
authors.lastname: Sakaki
mesh_terms: D015652:25-Hydroxyvitamin D 2 / Q000096:biosynthesis* / Q000378:metabolism; D002112:Calcifediol / Q000378:metabolism; D002117:Calcitriol / Q000378:metabolism; D053493:Cholestanetriol 26-Monooxygenase / Q000235:genetics / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid; D000072467:Cytochrome P450 Family 2; D057075:Enzyme Assays; D004875:Ergosterol / Q000378:metabolism; D005947:Glucose / Q000378:metabolism; D006801:Humans; D006900:Hydroxylation; D007536:Isomerism; D030781:Organisms, Genetically Modified / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism* / Q000528:radiation effects; D013997:Time Factors; D014466:Ultraviolet Rays; D047392:beta-Cyclodextrins / Q000378:metabolism
chemical_list: D047392:beta-Cyclodextrins; D015652:25-Hydroxyvitamin D 2; D000072467:Cytochrome P450 Family 2; C506833:CYP2R1 protein, human; D053493:Cholestanetriol 26-Monooxygenase; D002117:Calcitriol; D005947:Glucose; C031215:betadex; D002112:Calcifediol; D004875:Ergosterol