人参皂苷酶Ⅲ型转化人参皂苷Rc制备稀有皂苷C-Mc

摘要

利用基因构建克隆的E.coli C41菌产人参皂苷酶Ⅲ型,研究了酶转化人参皂苷Rc制备高活性稀有皂苷C-Mc的方法.结果发现,人参皂苷酶Ⅲ型先水解人参皂苷Rc的3-O-末端葡萄糖基生成C-Mc1,再水解C-Mc1的3-O-葡萄糖基生成C-Mc.人参皂苷酶Ⅲ型水解4.8 g的人参皂苷Rc,得到2.8 g产物,经HPLC检测,其纯度为90%.经核磁共振检测,产物结构为20-O-α-L-阿拉伯呋喃糖基-(1→6)-β-D-葡萄糖基-20(S)-二醇苷元,即C-Mc.%To study the enzymatic conversion from ginsenoside Rc to the rare ginsenoside C-Mc with high active, the ginsenosidase Ⅲ was expressed from recombinant Escherichia coli C41 and its enzymatic transformation processes were explored.The 3-O-glucopyranoside end of ginsenoside Rc was hydrolyzed to C-Mc1 by ginsenosidase Ⅲ, then C-Mc1 was transformed to C-Mc.4.8 g ginsenoside Rc was hydrolyzed to 2.8 g products by ginsenosidase Ⅲ, of which purity could reach to 90% determined by HPLC.The result of NMR showed that its structure was 20-O-[α-L-arabinofuranosyl-(1→6)-β-D-glucopyranosyl]-20(S)-protopanaxadiol, that was C-Mc.