| 重组嗜热β-葡萄糖苷酶转化稀有人参皂苷Rd和CK |
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| 引用本文： | 许春春,于渤浩,王红蕾,李晶,刘淑莹,于珊珊.重组嗜热β-葡萄糖苷酶转化稀有人参皂苷Rd和CK[J].高等学校化学学报,2016,37(2):281. |
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| 作者姓名： | 许春春 于渤浩 王红蕾 李晶 刘淑莹 于珊珊 |
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| 作者单位： | 1. 长春中医药大学吉林省人参科学研究院, 长春 1300002. 长春工业大学化学与生命科学学院, 长春 1300243. 中国科学院长春应用化学研究所长春质谱中心, 长春 1300224. 上海交通大学生命科学技术学院, 上海 200240 |
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| 基金项目： | 国家自然科学基金(批准号: 31400682)和长春市重大科技攻关计划项目(批准号: 13KG60)资助 |
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| 摘 要： | 利用高效液相色谱(HPLC)法, 对重组嗜热β-葡萄糖苷酶(Fpglu1)转化稀有人参皂苷(Rd和CK)进行研究, 并表征了其催化动力学参数. 利用同源模建和分子动力学模拟等生物信息学技术, 探究了Fpglu1转化人参皂苷的结构基础及其相互作用. 结果表明, Fpglu1能够水解人参总皂苷生成稀有皂苷Rd和CK, 其催化人参皂苷Rb1, Rb2和Rc的Km值分别为0.318, 1.840和5.269 mmol/L; 酶的转换数(kcat)值分别为144.191, 0.572和0.011 s-1. 当转化时间分别为6和102 h时, Rd和CK的产率达到最大, 分别为60%和93%. 通过对该酶的结构预测及皂苷分子的对接研究发现, 底物位于由疏水性氨基酸构成的底物口袋中, 氨基酸残基Glu194和Glu367是参与催化作用的关键, 且实验测得的酶促反应动力学参数(Km)与对接的相互作用能量值存在线性关系.
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| 关 键 词： | β-葡萄糖苷酶 生物转化 稀有人参皂苷 动力学 |
| 收稿时间： | 2015-07-09 |
Transformation of Minor Ginsenoside Rd and CK by Recombinant Thermostable β-Glucosidase† |
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| XU Chunchun,YU Bohao,WANG Honglei,LI Jing,LIU Shuying,YU Shanshan.Transformation of Minor Ginsenoside Rd and CK by Recombinant Thermostable β-Glucosidase†[J].Chemical Research In Chinese Universities,2016,37(2):281. |
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| Authors: | XU Chunchun YU Bohao WANG Honglei LI Jing LIU Shuying YU Shanshan |
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| Institution: | 1. Jilin Ginseng Academy, Changchun University of Chinese Medicine, Changchun 130000, China2. School of Chemistry and Bioscience, Changchun University of Technology, Changchun 130024, China3. Changchun Center of Mass Spectrum, Changchun Institute of Applied Chemistry,Chinese Academy of Sciences, Changchun 130022, China4. School of Life Sciences and Biotechnology, Shanghai Jiaotong University, Shanghai 200240, China |
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| Abstract: | Ginsenosides are the principle components that are responsible for the biological and pharmacological activities of ginseng. In this study, biotransformation of ginsenside Rd and CK by recombinant β-glucosidasse(Fpglu1) was studied by high performance liquid chromatography(HPLC). Structure modeling and molecular docking studies were performed to study the interaction between the protein and the ginsenosides. The results showed that ginsenoside Rd and CK could be transformed from major ginsenosides Rb1, Rb2 and Rc by Fpglu1. The Km values for Rb1, Rb2 and Rc were 0.318, 1.840 and 5.269 mmol/L, and the kcat values were 144.191, 0.572 and 0.011 s-1, respectively. The highest productivity of ginsenoside Rd and CK were 60% and 93% with biotransformation time of 6 and 102 h, respectively. Molecular docking studies show that Glu194 and Glu367 are key active site residues and the predicted inter-energy exhibits linear relations to experimental Km values and kcat values. |
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| Keywords: | β-Glucosidase Biotransformation Ginsenoside Kinetics |
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