title: A strategy for enrichment of the bioactive sphingoid base-1-phosphates produced by Hypericum perforatum L. in a balloon type airlift reactor.
abstract: An efficient enrichment method using immobilized metal affinity chromatography (IMAC) was developed for selective extraction of bioactive sphingoid base-1-phosphates (SB1Ps) from adventitious roots of Hypericum perforatum cultured in bioreactor. The phosphate-selective IMAC enrichment coupled with LC-MS/MS enabled sensitive analysis of low-abundance SB1Ps present in the root biomass, which would not be feasible otherwise due to severe interferences from complex biological matrices. The time-dependent growth rate and production of SB1Ps from adventitious roots were investigated. The level of phytosphingosine-1-phosphate, which was found to be the major SB1Ps, reached a maximum amount of 635.6pmolpergram of dry weight after 3weeks of culture and decreased between 3 and 5weeks of culture subsequently. On the other hand, sphingosine-1-phosphate and sphinganine-1-phosphate were present at levels of 18.91 and 73.15pmolpergram of dry weight, respectively, after a week of culture and their level decreased thereafter.
authors.affiliation: Center for Analytical Chemistry, Division of Metrology for Quality of Life, Korea.
authors.forename: You-Xun
authors.initials: YX
authors.lastname: Jin
authors.forename: Xi-Hua
authors.initials: XH
authors.lastname: Cui
authors.forename: Kee-Yoeup
authors.initials: KY
authors.lastname: Paek
authors.forename: Yong-Hyeon
authors.initials: YH
authors.lastname: Yim
mesh_terms: D019149:Bioreactors; D001709:Biotechnology / Q000295:instrumentation* / Q000379:methods*; D002725:Chloroform / Q000737:chemistry; D002846:Chromatography, Affinity; D002851:Chromatography, High Pressure Liquid; D020902:Hypericum / Q000254:growth & development / Q000378:metabolism*; D008246:Lysophospholipids / Q000096:biosynthesis*; D013058:Mass Spectrometry; D008670:Metals / Q000737:chemistry; D000432:Methanol / Q000737:chemistry; D018517:Plant Roots / Q000254:growth & development / Q000378:metabolism; D012997:Solvents / Q000737:chemistry; D013110:Sphingosine / Q000031:analogs & derivatives* / Q000096:biosynthesis; D013997:Time Factors
chemical_list: D008246:Lysophospholipids; D008670:Metals; D012997:Solvents; C117422:phytosphingosine-1-phosphate; C060504:dihydrosphingosine 1-phosphate; C060506:sphingosine 1-phosphate; D002725:Chloroform; D013110:Sphingosine; D000432:Methanol