Efficient production of the anti-aging drug Cycloastragenol: insight from two Glycosidases by enzyme mining
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BIOTECHNOLOGICAL PRODUCTS AND PROCESS ENGINEERING
Efficient production of the anti-aging drug Cycloastragenol: insight from two Glycosidases by enzyme mining Leiyu Cheng 1,2 & Han Zhang 1 & Haiyang Cui 3 & Wenya Wang 2,4 & Qipeng Yuan 1,2 Received: 2 September 2020 / Revised: 9 October 2020 / Accepted: 18 October 2020 # Springer-Verlag GmbH Germany, part of Springer Nature 2020
Abstract The telomerase activator cycloastragenol (CA) is regarded as a potential anti-aging drug with promising applications in the food and medical industry. However, one remaining challenge is the low efficiency of CA production. Herein, we developed an enzyme-based approach by applying two enzymes (β-xylosidase: Xyl-T; β-glucosidase: Bgcm) for efficient CA production. Both key glycosidases, mined by activity tracking or homology sequence screening, were successfully over-expressed and showed prominent enzymatic activity profiles, including widely pH stability (Xyl-T: pH 3.0–8.0; Bgcm: pH 4.0–10.0), high catalytic efficiency (kcat/Km: 0.096 mM-1s−1 (Xyl-T) and 3.08 mM-1s−1 (Bgcm)), and mesophilic optimum catalytic temperature (50 °C). Besides, the putative catalytic residues (Xyl-T: Asp311/Glu 521; Bgcm: Asp311/Glu 521) and the potential substratebinding mechanism of Xyl-T and Bgcm were predicted by comprehensive computational analysis, providing valuable insight into the hydrolysis of substrates at the molecular level. Notably, a rationally designed two-step reaction process was introduced to improve the CA yield and increased up to 96.5% in the gram-scale production, providing a potential alternative for the industrial CA bio-production. In essence, the explored enzymes, the developed enzyme-based approach, and the obtained knowledge from catalytic mechanisms empower researchers to further engineer the CA production and might be applied for other chemicals synthesis. Key points • A β-xylosidase and a β-glucosidase were mined to hydrolyze ASI into CA. • The two recombinant glycosidases showed prominent catalytic profiles. • Two-step enzymatic catalysis for CA production from ASI was developed.
Keywords Cycloastragenol . β-Xylosidase . β-Glucosidase . Two-step enzymatic catalysis
Supplementary Information The online version contains supplementary material available at https://doi.org/10.1007/s00253-02010966-5. * Wenya Wang [email protected]
2
State Key Laboratory of Chemical Resource Engineering, College of Life Science and Technology, Beijing University of Chemical Technology, No.15 North Third Ring East Road, Chao yang District, Beijing 100029, People’s Republic of China
3
Institute of Biotechnology, RWTH Aachen University, Worringerweg 3, 52074 Aachen, Germany
4
Amoy-BUCT Industrial Bio-technovation Institute, Beijing University of Chemical Technology, No.126 Xinyuan Road, Haicang District, Amoy 361028, People’s Republic of China
* Qipeng Yuan [email protected] 1
Beijing Advanced Innovation Center for Soft Matter Science and Engineering, Beijing University of Chemical Technology, No.15 North Third Ring East Road
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