Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Icaritin Preparation from Icariin by a Special Epimedium Flavonoid-Glycosidase from Aspergillus sp.y848 Strain

  • Wang, Zhenghao (College of Biotechnology, Dalian Polytechnic University) ;
  • Liu, Chunying (School of Life Science and Biotechnology, Liaoning Marine Microbial Engineering and Technology Center, Dalian University) ;
  • Yu, Hongshan (College of Biotechnology, Dalian Polytechnic University) ;
  • Wu, Bo (College of Biotechnology, Dalian Polytechnic University) ;
  • Huai, Baoyu (College of Biotechnology, Dalian Polytechnic University) ;
  • Zhuang, Ziyu (Dalian Center for Certification and Food and Drug) ;
  • Sun, Changkai (Research & Educational Center for the Control Engineering of Translational Precision Medicine, Dalian University of Technology) ;
  • Xu, Longquan (College of Biotechnology, Dalian Polytechnic University) ;
  • Jin, Fengxie (College of Biotechnology, Dalian Polytechnic University)
  • Received : 2021.12.20
  • Accepted : 2022.02.09
  • Published : 2022.04.28
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Abstract

In this study, to obtain icaritin with high pharmacological activities from icariin, which has a content ratio of over 58% in the total flavonoids of Epimedium herb, a special Epimedium flavonoid-glycosidase was produced, purified and characterized from Aspergillus sp.y848 strain. The optimal enzyme production was gained in a medium containing 5% (w/v) wheat bran extract and 0.7% (w/v) Epimedium leaf powder as the enzyme inducer, and strain culture at 30℃ for 6-7 days. The molecular weight of the enzyme was approximately 73.2 kDa; the optimal pH and temperature were 5.0 and 40℃. The enzyme Km and Vmax values for icariin were 15.63 mM and 55.56 mM/h. Moreover, the enzyme hydrolyzed the 7-O-glucosides of icariin into icariside II, and finally hydrolyzed 3-Orhamnoside of icariside II into icaritin. The enzyme also hydrolyzed 7-O-glucosides of epimedin B to sagittatoside B, and then further hydrolyzed terminal 3-O-xyloside of sagittatoside B to icarisiede II, before finally hydrolyzing 3-O-rhamnoside of icarisiede II into icaritin. The enzyme only hydrolyzed 7-O-glucoside of epimedin A or epimedin C into sagittatoside A or sagittatoside C. It is possible to prepare icaritin from the high-content icariin in Epimedium herb using this enzyme. When 2.5% icariin was reacted at 40℃ for 18-20 h by the low-cost crude enzyme, 5.04 g icaritin with 98% purity was obtained from 10 g icariin. Also, the icaritin molar yield was 92.5%. Our results showed icaritin was successfully produced via cost-effective and relatively simple methods from icariin by crude enzyme. Our results should be very useful for the development of medicines from Epimedium herb.

Keywords

Acknowledgement

This work was supported by the National Key Research and Development Program of China, No. 2018AAA0100300 and 2012ZX09503001-003, and the National High-end Foreign Expert Project of China, No. GDT20152100019. The NMR experiment was performed and analyzed by senior engineer Yijie Wu from the Changchun Institute of Applied Chemistry Chinese Academy of Sciences, People's Republic of China.

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