Generation of recombinant destabilase-lysozyme from medicinal leeches in three different expression systems
Recombinant expression study published in 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.
Resumen
Comparative recombinant expression of leech destabilase-lysozyme in E. coli, P. pastoris, and N. tabacum (plant) systems. Plant expression provides correctly folded, glycosylated protein.
Por qué esto importa para la hirudoterapia
Este estudio detalla la generación de la isoforma 2 recombinante de desestabilasa-lisozima, una enzima multifuncional de sanguijuelas medicinales, utilizando tres sistemas de expresión diferentes: E. coli, P. pastoris y la línea celular humana Expi293F. Los investigadores compararon los sistemas y hallaron que las enzimas producidas en formas solubles poseían actividades lisozima, isopeptidasa, fibrinolítica y antimicrobiana significativamente mayores que las replegadas a partir de cuerpos de inclusión. Para ASH, esto es prácticamente relevante, ya que identifica los métodos biotecnológicos óptimos para producir enzimas terapéuticas funcionales derivadas de la sanguijuela. La salvedad es que se trata estrictamente de un estudio de producción y caracterización bioquímica, sin datos in vivo ni clínicos que demuestren la eficacia terapéutica.
Citación
Generation of recombinant destabilase-lysozyme from medicinal leeches in three different expression systems.
Manuvera VA et al. · Protein expression and purification, 2015
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Añadido a la biblioteca ASH: May 27, 2026 · Última actualización del sitio: 18 de junio de 2026