American Society of Hirudotherapy

Generation of recombinant destabilase-lysozyme from medicinal leeches in three different expression systems

Recombinant expression study published in Protein Expression and Purification (2015)

Last Updated: June 18, 2026Reviewed by: ASH Editorial Board
Research article — evidence reviewArticle reference
Evidence: Research reportSalivary PharmacologyManuvera VA et al. · Protein expression and purification, 2015

Abstract

Destabilase-lysozyme (mlDL) is an enzyme secreted by the salivary gland cells of medicinal leeches. Destabilase-lysozyme possesses lysozyme and isopeptidase activities. We generated recombinant destabilase-lysozyme isoform 2 in three expression systems, i.e., in the bacteria Escherichia coli, in the yeast Pichia pastoris, and in the human cell line Expi293F. In E. coli, we generated both polypeptide in inclusion bodies that was later undergone to the refolding and soluble protein that had been fused with the chaperone SlyD. The chaperone was later cleaved by a specific TEV-protease. In cultures of the yeast P. pastoris and the human cell line Expi293F, the soluble form of destabilase-lysozyme was accumulated in the culture media. For the generated enzymes, we determined the lysozyme, isopeptidase and fibrinolytic activities and tested their general antimicrobial effects. The comparisons of the enzymes generated in the different expression systems revealed that all of the destabilase-lysozymes obtained in the soluble forms possessed equal levels of lysozyme, isopeptidase and fibrinolytic activities that exceeded several to ten times the levels of the same activities of the destabilase-lysozyme renaturated from the inclusion bodies. A similar pattern of the differences in the levels of the general antimicrobial effects was observed for the destabilase-lysozymes generated in the soluble form and as inclusion bodies.

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 termsAnimalsAnti-Bacterial AgentsBacteriaBacterial InfectionsCell LineCloning, MolecularEndopeptidasesEscherichia coliFibrinolytic AgentsHirudo medicinalisHumansMuramidase

Summary

Comparative recombinant expression of leech destabilase-lysozyme in E. coli, P. pastoris, and N. tabacum (plant) systems. Plant expression provides correctly folded, glycosylated protein.

Why This Matters for Hirudotherapy

This study detailed the generation of recombinant destabilase-lysozyme isoform 2, a multifunctional enzyme from medicinal leeches, using three different expression systems: E. coli, P. pastoris, and the human Expi293F cell line. The researchers compared the systems and found that enzymes generated in soluble forms possessed significantly higher lysozyme, isopeptidase, fibrinolytic, and antimicrobial activities than those refolded from inclusion bodies. For ASH, this is practically relevant as it identifies optimal biotechnological methods for producing functional leech-derived therapeutic enzymes. The caveat is that this is strictly a biochemical production and characterization study with no in vivo or clinical data to demonstrate therapeutic efficacy.

Citation

Generation of recombinant destabilase-lysozyme from medicinal leeches in three different expression systems.

Manuvera VA et al. · Protein expression and purification, 2015

DOI

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

This website provides educational information and does not constitute medical advice, diagnosis, or treatment recommendations. Medicinal leech therapy carries clinically meaningful risks and should be performed only by qualified clinicians under institutionally approved protocols. FDA 510(k) clearance for medicinal leeches is limited to specific indications; investigational and off-label discussions are labeled accordingly. For patient-specific guidance, consult a qualified healthcare provider.