新型双相酶水解体系制备宝藿苷Ⅰ
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摘要
目的构建新型双相酶水解体系制备宝藿苷Ⅰ。方法单因素试验优化缓冲液pH值、酶与底物质量比、酶解时间、反应温度,并在淫羊藿苷处理量、酶重复使用、操作方便性方面上与传统酶水解法进行比较。结果在醋酸-醋酸钠缓冲液(pH 4. 0)和乙酸乙酯构建的双相酶水解体系中,当β-葡萄糖苷酶与淫羊藿苷的质量比为1∶1时,于55℃下反应6 h后水解率达到100%,双相水解法可将淫羊藿苷处理量提高到60 mg/mL,是传统酶水解法的2倍,并且制备步骤得以简化。此外,双相酶水解体系中酶液使用2次后,淫羊藿苷转化率仍能达到98. 58%。结论该方法可高效、便捷地水解淫羊藿苷以获取宝藿苷Ⅰ,并实现了酶的重复利用。
引文
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[3] Wu Y Q,Cao L Y,Xia L G,et al. Evaluation of osteogenesis and angiogenesis of icariin in local controlled release and systemic delivery for calvarial defect in ovariectomized rats[J]. Sci Rep,2017,7(1):5077.
[4] Tan H L,Chan K G,Pusparajah P,et al. Anti-cancer properties of the naturally occurring aphrodisiacs:icariin and its derivatives[J]. Front Pharmacol,2016,7:191.
[5] Li C R,Li Q,Mei Q B,et al. Pharmacological effects and pharmacokinetic properties of icariin,the major bioactive component in Herba Epimedii[J]. Life Sci,2015,126:57-68.
[6] Cheng T,Zhang Y,Zhang T,et al. Comparative pharmacokinetics study of icariin and icarisideⅡin rats[J]. Molecules,2015,20(12):21274-21286.
[7] Wang G,Wang C Y,Zhang B J,et al. Rapid separation of flavonoids from hydrolysis products of Epimedium koreanum[J].J Liq Chromatogr Relat Technol,2013,36(9):1163-1176.
[8] Liu D F,Li Y P,Ou T M,et al. Synthesis and antimultidrug resistance evaluation of icariin and its derivatives[J]. Bioorg Med Chem Lett,2009,19(15):4237-4240.
[9] Wu H L,Kim M H,Han J H. Icariin metabolism by human intestinal microflora[J]. Molecules,2016,21(9):E1158.
[10]彭静,马益华,陈彦,等.固定化β-葡萄糖苷酶和蜗牛酶生物转化淫羊藿苷效率的比较研究[J].药学学报,2015,50(12):1652-1659.
[11]彭静,马益华,陈彦,等.微乳化-凝胶法制备微球固定化蜗牛酶并生物转化淫羊藿苷研究[J].中草药,2015,46(22):3326-3332.
[12] Pozharitskaya O N,Kosman V M,Shikov A N,et al. Comparison between HPLC and HPTLC densitometry for the determination of icariin from Epimedium koreanum extracts[J]. J Sep Sci,2007,30(5):708-712.
[13] A2ic A,Be2ic L,Muhovic I,et al. Purification and characterization ofβ-glucosidase from Agaricus bisporus(white button mushroom)[J]. Protein J,2015,34(6):453-461.
[14]贾东升,贾晓斌,赵江丽,等.纤维素酶转化淫羊藿苷制备宝藿苷Ⅰ的研究[J].中草药,2010,41(6):888-892.