American Society of Hirudotherapy

Cloning, characterization, and heterologous expression of a candidate hirudin gene from the salivary gland transcriptome of Hirudo nipponia

Recombinant expression study published in Scientific Reports (2023)

Last Updated: June 18, 2026Reviewed by: ASH Editorial Board
Research article — evidence reviewArticle reference
Evidence: In vitro / laboratoryGenomics & ProteomicsSalivary PharmacologyDrug DevelopmentShi P et al. · Scientific reports, 2023

Abstract

Hirudin is a pharmacologically active substance in leeches with potent blood anticoagulation properties. Although recombinant hirudin production isolated from Hirudo medicinalis Linnaeus and Hirudinaria manillensis Lesson is known, to our knowledge, this study is the first to report recombinant hirudin expression and production from Hirudo nipponia Whitman. Thus, the present study aimed to clone and characterize the full-length cDNA of a candidate hirudin gene (c16237_g1), which is localized on the salivary gland transcriptome of H. nipponia, and further evaluate its recombinant production using a eukaryotic expression system. The 489-bp cDNA possessed several properties of the hirudin "core" motifs associated with binding to the thrombin catalytic pocket. A fusion expression vector (pPIC9K-hirudin) was constructed and successfully transformed into Pichia pastoris strain GS115 via electroporation. Sodium dodecyl sulphate-polyacrylamide gel electrophoresis and western blot analysis confirmed hirudin expression. The recombinant protein was expressed with a yield of 6.68 mg/L culture. Mass spectrometry analysis further confirmed target protein expression. The concentration and antithrombin activity of purified hirudin were 1.67 mg/mL and 14,000 ATU/mL, respectively. These findings provide a basis for further elucidating the molecular anticoagulation mechanism of hirudin, and address China's growing market demand for engineered H. nipponia-derived hirudin and hirudin-based drugs.

Abstract sourced from PubMed (NCBI) for the cited record. See the original publication for the authoritative version.

Publication typeJournal ArticleResearch Support, Non-U.S. Gov't
Indexed MeSH termsAnimalsHirudinsAmino Acid SequenceDNA, ComplementaryTranscriptomeLeechesAnticoagulantsRecombinant ProteinsCloning, Molecular

Summary

Identifies and recombinantly expresses a hirudin variant from Hirudo nipponia (Asian medicinal leech) — characterizes thrombin inhibition kinetics and structural features.

Why This Matters for Hirudotherapy

Researchers cloned and characterized a candidate hirudin gene from the salivary gland transcriptome of the medicinal leech Hirudo nipponia, achieving successful heterologous expression of the recombinant protein. The purified recombinant hirudin demonstrated potent antithrombin activity, establishing a viable biotechnological method to produce this key anticoagulant. For ASH's domain, this study directly addresses the growing pharmaceutical demand for engineered, leech-derived anticoagulants and elucidates the molecular mechanisms of hirudin. The main caveat is that the findings represent strictly preclinical, in-vitro biomanufacturing data; the abstract provides no clinical trials or in-vivo efficacy data for this specific H. nipponia derivative.

Citation

Cloning, characterization, and heterologous expression of a candidate hirudin gene from the salivary gland transcriptome of Hirudo nipponia.

Shi P et al. · Scientific reports, 2023

DOI

Added to ASH library: May 27, 2026 · Site last updated: June 18, 2026

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