American Society of Hirudotherapy

Recombinant destabilase-lysozyme: synthesis de novo in E. coli and action mechanism of the enzyme expressed in Spodoptera frugiperda

Basic science published in Biochem Mosc (2004)

Last Updated: June 18, 2026Reviewed by: ASH Editorial Board
Research article — evidence reviewArticle reference
Evidence: In vitro / laboratoryDrug DevelopmentGenomics & ProteomicsZavalova LL et al. · Biochemistry (Mosc), 2004

Abstract

Destabilase-lysozyme (DL) from salivary gland secretion of the medicinal leech (Hirudo medicinalis) is as a member of the invertebrate lysozyme family, which sharply differs from other lysozyme families. In this study, DL lysozyme function was confirmed during expression of a gene encoding DL in Escherichia coli. Several constructs of the expression vectors pKK OmpA and pET-3A with or without bacterial, leech, or yeast signal peptides (SP) were used. The use of a construct without signal peptide genes resulted in normal growth of the transformed cells. Transformation of E. coli cells with the constructs containing SP was accompanied by the disruption of the forming cells. The use of the expression vector pET-32 LTC-System for production of DL as a fusion protein with thioredoxin also resulted in normal cell growth. However, specific activity of DL isolated from such cells was significantly lower than that of enzyme purified from extracts of Spodoptera frugiperda cells, which were infected with the baculovirus vector carrying DL cDNA. It is shown that the action mechanism of invertebrate lysozyme does not differ from that of other families: recombinant DL from S. frugiperda extracts catalyzed cleavage of synthetic substrate, hexamer of N-acetylglucosamine, to di- and tetramers, which is typical for enzymatic function of other lysozyme families.

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 termsAcetylglucosamineAnimalsCatalysisChromatography, AffinityEndopeptidasesEscherichia coli ProteinsHirudo medicinalisMuramidaseRecombinant ProteinsSpodopteraSubstrate Specificity

Summary

Functional recombinant destabilase-lysozyme produced in E. coli and S. frugiperda systems shows lysozyme catalysis cleaving N-acetylglucosamine hexamers.

Why This Matters for Hirudotherapy

This study examined the recombinant synthesis and action mechanism of destabilase-lysozyme (DL), an invertebrate lysozyme originally derived from the salivary gland secretion of the medicinal leech (Hirudo medicinalis). It is highly relevant to characterizing the leech secretome, confirming that DL's enzymatic action mechanism functions similarly to other lysozyme families by cleaving synthetic substrates into di- and tetramers. The research successfully expressed the leech enzyme in Escherichia coli and Spodoptera frugiperda cells to study its specific catalytic activity. The clinical relevance to live hirudotherapy is limited, as the focus is strictly on the molecular biology, heterologous expression, and in vitro biochemical mechanism of an isolated leech protein.

Citation

Recombinant destabilase-lysozyme: synthesis de novo in E. coli and action mechanism of the enzyme expressed in Spodoptera frugiperda.

Zavalova LL et al. · Biochemistry (Mosc), 2004

DOI

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

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