Optimization of Reynoutria sachalinensis leaf extract preparation for determination of shikimate dehydrogenase activity
- 作者: Bondarev S.A.1, Sereda A.A.1, Baleev D.N.1, Nikonorova E.R.1
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隶属关系:
- All-Russian Scientific Research Institute of Medicinal and Aromatic Plants
- 期: 卷 28, 编号 10 (2025)
- 页面: 50-56
- 栏目: Pharmaceutical chemistry
- URL: https://journal-vniispk.ru/1560-9596/article/view/334355
- DOI: https://doi.org/10.29296/25877313-2025-10-07
- ID: 334355
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详细
Introduction. Reynoutria sachalinensis is a promising subject for studying the mechanisms of tannin synthesis in plants due to the high proanthocyanidins content.
This study aimed to optimize the preparation of R. sachalinensis leaf extracts to determine the activity of shikimate dehydrogenase, a key enzyme in the shikimate pathway involved in tannin biosynthesis.
Material and methods. Aqueous extraction followed by protein precipitation using 60 or 80% (v/v) acetone, with or without glycerol, was carried out. The effect of freezing at –80 °C on shikimate dehydrogenase activity was also assessed.
Results. It was found that freezing both crude and acetone-purified extracts did not decrease shikimate dehydrogenase activity, while increasing the acetone concentration to 80% (v/v) led to an increase in enzyme activity compared to 60%(v/v) acetone. Glycerol had no significant effect on enzyme activity.
Conclusion. Thus, the optimized extraction using 80% (v/v) acetone precipitation and low-temperature storage allows for efficient extraction of proteins and enzymes, particularly shikimate dehydrogenase, from R. sachalinensis leaves for subsequent biochemical studies.
作者简介
S. Bondarev
All-Russian Scientific Research Institute of Medicinal and Aromatic Plants
编辑信件的主要联系方式.
Email: xcsergey@mail.ru
ORCID iD: 0009-0009-7039-2939
SPIN 代码: 9186-5325
Junior Researcher, Laboratory of Genomics and Biochemistry
俄罗斯联邦, 7 Green str., Moscow, 117216A. Sereda
All-Russian Scientific Research Institute of Medicinal and Aromatic Plants
Email: nensi.sereda@mail.ru
ORCID iD: 0009-0007-1891-4846
SPIN 代码: 6292-5126
Junior Researcher, Laboratory of Genomics and Biochemistry of Medicinal Plants
俄罗斯联邦, 7 Green str., Moscow, 117216D. Baleev
All-Russian Scientific Research Institute of Medicinal and Aromatic Plants
Email: dbaleev@gmail.com
ORCID iD: 0000-0002-1228-0594
Ph.D. (Agrical.), Leading Researcher, Laboratory of Atomic-Molecular Bioregulation and Selection
俄罗斯联邦, 7 Green str., Moscow, 117216E. Nikonorova
All-Russian Scientific Research Institute of Medicinal and Aromatic Plants
Email: gatiatulinaer@gmail.com
ORCID iD: 0000-0002-6360-2194
SPIN 代码: 5392-5170
Ph.D. (Med.), Leading Researcher, Laboratory of Genomics and Biochemistry of Medicinal Plants
俄罗斯联邦, 7 Green str., Moscow, 117216参考
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