基于氯化胆碱的深共晶溶剂中通过酶反应介质转化人参皂苷。
Transformation of ginsenoside via deep eutectic solvents based on choline chloride as an enzymatic reaction medium.
机构信息
Shaanxi Key Laboratory of Degradable Biomedical Materials, School of Chemical Engineering, Northwest University, Taibai North Road 229, Xi'an, 710069, Shaanxi, China.
Shaanxi R&D Center of Biomaterials and Fermentation Engineering, School of Chemical Engineering, Northwest University, Taibai North Road 229, Xi'an, 710069, Shaanxi, China.
出版信息
Bioprocess Biosyst Eng. 2020 Jul;43(7):1195-1208. doi: 10.1007/s00449-020-02314-8. Epub 2020 Mar 5.
Ginsenoside compound K (CK) with a wide range of pharmacological activities has been widely used in the healthcare product industry. However, the application of CK is limited by low productivity and difficult separation. The purpose of this study is to convert ginsenoside Rb1 into CK by improving conversion efficiency in novel "green" reaction medium-deep eutectic solvent (DES). Talaromyces purpureogenus was selected from ginseng rhizosphere soil to produce β-glucosidase with high activity and purity to transform ginsenosides, and Mn was found to be an enzyme promoter. Among the DES based on choline chloride as hydrogen-bond receptor, choline chloride:ethylene glycol (ChCl:EG = 2:1) was the most promising solvent in maintaining enzyme activity and stability. In the presence of 30% v/v ChCl:EG = 2:1, the half-life of β-glucosidase was increased by 96%, the solubility of F2 was increased by 120%, and CK yield was increased by 54% compared with those in the buffer. Fourier transform infrared, circular dichroism, and fluorescence spectroscopy confirmed that DES did not destroy the structure and conformation of β-glucosidase. In addition, 80.6% CK conversion was obtained at 60 °C, pH 4.5, 48 h and 8 mM Rb1, which provided a feasible method for efficiently producing CK.
人参皂苷化合物 K（CK）具有广泛的药理活性，已广泛应用于保健品行业。然而，由于产量低、分离困难，CK 的应用受到限制。本研究旨在通过在新型“绿色”反应介质-深共熔溶剂（DES）中提高转化效率，将人参皂苷 Rb1 转化为 CK。从人参根际土壤中筛选出紫色红曲霉菌，以生产具有高活性和高纯度的β-葡萄糖苷酶来转化人参皂苷，发现 Mn 是一种酶促剂。在以氯化胆碱为氢键受体的 DES 中，氯化胆碱：乙二醇（ChCl：EG=2：1）是最有前途的溶剂，既能保持酶的活性又能保持酶的稳定性。在 30%v/v ChCl：EG=2：1 的存在下，β-葡萄糖苷酶的半衰期延长了 96%，F2 的溶解度提高了 120%，CK 的产率比缓冲液中的提高了 54%。傅里叶变换红外、圆二色和荧光光谱证实 DES 没有破坏β-葡萄糖苷酶的结构和构象。此外，在 60°C、pH4.5、48 小时和 8mM Rb1 条件下，获得了 80.6%的 CK 转化率，为高效生产 CK 提供了一种可行的方法。
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