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Citation: SUN Hong-Mei, LU Ze-Lin, ZHAO Huan-Xi, ZHENG Fei, LI Jing, LIU Ming, YUE Hao, YU Shan-Shan. Liquid Chromatography-Mass Spectrometry-Directed Evolution Enzyme Combination Technique for Investigation of Triterpenoid Saponin Transformation[J]. Chinese Journal of Analytical Chemistry, ;2020, 48(1): 129-136. doi: 10.19756/j.issn.0253-3820.191501 shu

Liquid Chromatography-Mass Spectrometry-Directed Evolution Enzyme Combination Technique for Investigation of Triterpenoid Saponin Transformation

Jilin Ginseng Academy, Changchun University of Chinese Medicine, Changchun 130117, China

Received Date: 15 August 2019
Revised Date: 8 November 2019

Fund Project: This work was supported by the Jilin Province Science and Technology Development Plan Project (No.20170307029YY) and the Jilin Province Department of Education "13th Five-year" Scientific Program (No. JJKH2018125K).

The directed evolution technology was used for recombination of glycosidase ABQ to transform triterpenoid saponins with different structures, including Esculentoside A, Notoginsenoside R₁ and Saikosaponin A, and the biotransformation process of the triterpenoid saponins was investigated by rapid resolution liquid chromatography coupled with quadruple-time-of-flight mass spectrometry (RRLC/Q-TOF-MS). The result showed that ABQ had high activities for Esculentoside A, Notoginsenoside R₁, Saikosaponin A, however, it showed different substrates selectivity for the three saponins. Besides, ABQ could catalyze the hydrolysis of C-3 lateral glucose of Esculentoside A and the C-20 lateral glucose of Notoginsenoside R₁ to produce Esculentoside B and Notoginsenoside R₂, repectively. And the recombinant glycosidase ABQ could catalyze the hydrolysis of C-3 lateral glucose of Saikosaponin A to produce Prosaikogenin F and then further catalyzed the hydrolysis of C-3 fucoside to produce Saikogenin F. In brief, the directed evolution technology could improve enzyme activity and environmental adaptability, and the chromatography-mass spectrometry could be used to evaluate the role of recombinant enzymes by real-time detection of prototypes and products.
- Triterpenoid saponins,
- Biotransformation,
- Liquid Chromatography-mass spectrometry,
- Enzyme directed evolution
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