Purification and characterization of an anticoagulant oligopeptide from Whitmania pigra Whitman

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Abstract
Pharmacognosy Magazine,2015,11,43,444-448.
Published:July 2015
Type:Original Article
Authors:
Author(s) affiliations:

Xiaobei Zheng1, Juan Li2, Zhengwang Chen3, Yimei Liu2, Keli Chen2
1 Department of Identification and Assessment of TCM, Hubei University of Traditional Chinese Medicine, Key Laboratory of TCM Resource and TCM Compound Co-constructed by Hubei province and Ministry of Education, New products of TCM Senile Diseases Co-Innovation Center of Hubei; Department of Quality Control, Wuhan Institute of Biological Products Co., Ltd, China
2 Department of Identification and Assessment of TCM, Hubei University of Traditional Chinese Medicine, Key Laboratory of TCM Resource and TCM Compound Co-constructed by Hubei province and Ministry of Education, New products of TCM Senile Diseases Co-Innovation Center of Hubei, China
3 Key Laboratory of Molecular Biophysics of the Ministry of Education, College of Life Science and Technology, Huazhong University of Science and Technology, Wuhan 430065, China

Abstract:

Background: Dried Whitmania pigra is used for the treatment of cardiovascular and cerebrovascular diseases in traditional Chinese medicine and hot water and alcohol extracts also have anticogulant activity. However, a lower molecular weight and more stable anticogulant is needed. Objective: The objective of the following study is to purify and characterize of an anticoagulant oligopeptide from Hirudo (Whitmania pigra Whitman). Materials and Methods: Gel filtration on Sephadex G 50, ion exchange on diethylaminoethyl cellulose, and semi prepared high performance liquid chromatography were used to purify Hirudo. Automated coagulation analyzer was used for evaluating anticoagulant activity. Molecular weight was measured by Matrix assisted laser desorption ionization time of flight mass spectrometry. Amino acid sequence of the oligopeptide was measured by amino acid sequence analyzer. Results: A new anticoagulant, named whitide, isolated from Hirudo was purified, with a molecular weight 1997.1 Da. Amino acid sequence of the oligopeptide was identified as Gly-Pro-ALa-Gly-Hyp-Val-Gly-Ala-Hyp-Gly-Gly-Hyp-Gly-Val-Arg-Gly-Leu-Hyp-Gly-Asp-Arg-Gly. The results revealed that its amino acid sequence had strong homology to various types of collagen. Conclusion: Whitide might be an orally anticoagulant for its hot and trypsin stable.

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