基于氯化胆碱的低共熔溶剂中采用底物补料分批策略生物催化生产化合物K。
Biocatalytic production of compound K in a deep eutectic solvent based on choline chloride using a substrate fed-batch strategy.
作者信息
Han Xin, Li Weina, Duan Zhiguang, Ma Xiaoxuan, Fan Daidi
机构信息
Shaanxi Key Laboratory of Degradable Biomedical Materials, School of Chemical Engineering, Northwest University, 229 North Taibai Road, Xi'an, Shaanxi 710069, China; Shaanxi R&D Center of Biomaterials and Fermentation Engineering, School of Chemical Engineering, Northwest University, 229 North Taibai Road, Xi'an, Shaanxi 710069, China; Biotech. & Biomed. Research Institute, Northwest University, 229 North Taibai Road, Xi'an, Shaanxi 710069, China.
Shaanxi Key Laboratory of Degradable Biomedical Materials, School of Chemical Engineering, Northwest University, 229 North Taibai Road, Xi'an, Shaanxi 710069, China; Shaanxi R&D Center of Biomaterials and Fermentation Engineering, School of Chemical Engineering, Northwest University, 229 North Taibai Road, Xi'an, Shaanxi 710069, China; Biotech. & Biomed. Research Institute, Northwest University, 229 North Taibai Road, Xi'an, Shaanxi 710069, China; Xi'an Giant Biogene Co., Ltd, Xi'an, Shaanxi 710065, China.
出版信息
Bioresour Technol. 2020 Jun;305:123039. doi: 10.1016/j.biortech.2020.123039. Epub 2020 Feb 19.
This study involved the development of a β-glucosidase-catalyzed hydrolysis method based on a deep eutectic solvent (DES), choline chloride-ethylene glycol 2:1, and continuous feed technique to overcome the difficulty of high-concentration ginsenoside hydrolysis. A productivity of 142 mg·L·h was achieved with the following conditions: 30 vol% DES, pH 5.0, 55 °C, and substrate concentration of 12 mM. In the presence of DES, the affinity and catalytic efficiency of β-glucosidase to Rd increased by 49 and 64%, respectively, which promoted the continuation of hydrolysis. Moreover, conformation of β-glucosidase was mostly retained, as confirmed by spectral information. Through a combination of a substrate fed-batch technique to reduce the inhibitory effects of substrates and products, the CK conversion rate increased by 44% compared to traditional single-batch in pure buffer. This report describes a practical method for the continuous conversion of natural compounds through biological processes and solvent engineering.
本研究开发了一种基于低共熔溶剂（DES）氯化胆碱 - 乙二醇（2:1）的β - 葡萄糖苷酶催化水解方法以及连续进料技术，以克服高浓度人参皂苷水解的困难。在以下条件下实现了142 mg·L·h的生产率：30%（体积）的DES、pH 5.0、55°C以及12 mM的底物浓度。在DES存在下，β - 葡萄糖苷酶对Rd的亲和力和催化效率分别提高了49%和64%，这促进了水解的持续进行。此外，光谱信息证实β - 葡萄糖苷酶的构象大部分得以保留。通过结合底物补料分批技术以降低底物和产物的抑制作用，与纯缓冲液中的传统单批次方法相比，CK转化率提高了44%。本报告描述了一种通过生物过程和溶剂工程实现天然化合物连续转化的实用方法。
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