Biotransformation of 6-Deoxo Type Brassinosteroids in Yeast (Saccharomyces cerevisiae) WAT21 Cells

다국어 초록 (Multilingual Abstract)

We recently demonstrated that non-plant cells, yeast WAT21, produce a steroidal plant hormone, castasterone. To understand how castasterone is generated in WAT21 cells, deuteriumlabeled (26, 28-2H6) 6-deoxoteasterone, 6-deoxotyphasterol and 6-deoxocastasterone were fed to WAT21 cells, and their metabolites were analyzed by gas chromatography-mass spectrometry. When [2H6]-6-deoxoteasterone was fed, [2H6]-6-deoxo-3-dehydroteasterone and [2H6]-6-deoxotyphasterol were identified as metabolites.
When [2H6]-6-deoxotyphasterol was used, [2H6]-6-deoxoteasterone and [2H6]-6-deoxo-3-dehydroteasterone and [2H6]-6-deoxocastasterone were detected. When [2H6]-6-deoxocastasterone was added, [2H6]-castasterone was identified. Taken together, a biosynthetic sequence,6-deoxoteasterone↔6-deoxo-3-dehydroteasterone↔6-deoxotyphasterol →6-deoxocastasterone→castasterone seems to function to produce brassinosteroids in WAT21 cells. Coupled with the presence of a biosynthetic sequence, teasterone→3-dehydroteasterone↔ typhasterol →castasterone, this suggests that brassinosteroids in WAT21 cells were biosynthesized via the same kind of pathways established in plants. This study provides a clue for possible mass-production of brassinosteroids in non-plant cells, yeast WAT21 for commercial use in agriculture.

참고문헌 (Reference)

1 Bajguz A, "The chemical characteristic and distribution of brassinosteroids in plants" 62 : 1027-1046, 2003

2 Kelly SL, "Purification and reconstitution of activity of Saccharomycescerevisiae P450 61, a sterol Δ22-desaturase" 377 : 217-220, 1995

3 Bishop GJ, "Plants steroid hormones, brassinosteroids : Current highlights of molecular aspects on their synthesis/metabolism, transport, perception and response" 42 : 114-120, 2001

4 Turi TG, "Multiple regulatory elements control expression of the gene encoding the Saccharomyces cerevisiae cytochrome P450, lanosterol 14 alpha-demethylase(ERG11)" 267 : 2046-2056, 1992

5 Sakurai A, "In Brassinosteroids: Steroidal Plant Hormones, (1st ed)" Springer-Verlag 1999

6 Buchnan BB, "In Biochemistry&Molecular Biology of Plants, (1st ed.)" American Society of Plant Physiologists 2000

7 Tae-Wuk Kim, "Identification of Castasterone from a Yeast (Saccharomyces cerevisiae) Strain, WAT21" 대한화학회 31 (31): 1430-1432, 2010

8 Se-HwanJoo, "Identification and Transformation of Campestanol in Cultured Cells of Phaseolus vulgaris" 대한화학회 23 (23): 27-1038, 2002

9 Duan H, "Heterologous expression and strategies for encapsulation of membrane-localized plant P450s" 5 : 507-523, 2006

10 Mallory JC, "Dap1p, a heme-binding protein that regulates the cytochrome P450 protein Erg11p/Cyp51p in Saccharomyces cerevisiae" 25 : 1669-1679, 2005

11 Lees ND, "Cloning of the late genes in the ergosterol biosynthetic pathway of Saccharomyces cerevisiae" 30 : 221-226, 1995

12 Urban P, "Characterization of recombinant plant cinnamate 4-hydroxylase produced in yeast. Kinetic and spectral properties of the major plant P450 of the phenylpropanoid pathway" 222 : 843-850, 2005

13 Sang Cheul Lee, "Biosynthesis of Brassinosteroids in a Yeast (Saccharomyces cerevisiae) Cell, WAT21: Conversion of Teasterone to Castasterone" 대한화학회 32 (32): 332-334, 2011

14 Kim T-W, "Arabidopsis CYP85A2, a cytochrome P450, mediates the Baeyer-Villiger oxidation of castasterone to brassinolide in brassinosteroid biosynthesis" AMER SOC PLANT BIOLOGISTS 17 : 2397-2412, 200508

15 최성화, "A Double Mutant for the CYP85A1 and CYP85A2 Genes of Arabidopsis Exhibits a Brassinosteroid Dwarf Phenotype" 한국식물학회 48 (48): 237-244, 2005