Center for Analytical Chemistry, Division of Metrology for Quality of Life, Korea.
Bioresour Technol. 2012 Nov;123:284-9. doi: 10.1016/j.biortech.2012.07.042. Epub 2012 Jul 23.
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.
一种高效的富集方法，采用固定化金属亲和层析（IMAC），从生物反应器培养的贯叶连翘不定根中选择性提取生物活性神经鞘氨醇-1-磷酸（SB1P）。磷酸选择性 IMAC 富集与 LC-MS/MS 相结合，能够灵敏分析根生物质中低丰度的 SB1P，否则由于复杂生物基质的严重干扰，这将是不可能的。研究了不定根的生长速率和 SB1P 的产生随时间的变化。在培养 3 周后，发现植物鞘氨醇-1-磷酸的含量达到最大值，为每克干重 635.6pmol，随后在 3 至 5 周的培养过程中减少。另一方面，在培养 1 周后，神经鞘氨醇-1-磷酸和鲨肌醇-1-磷酸的含量分别为 18.91 和 73.15pmol/克干重，此后其含量下降。
