Antifungal and antibacterial functions of medicinal leech recombinant destabilase-lysozyme and its heated-up derivative

Abstract

Antifungal activity of recombinant medicinal leech destabilase-lysozyme (rec.Dest-Lys) was investigated by using fungi: Botrytis cinerea, and Verticillium lateriticum, including yeasts Candida guillermoudii and Shizosaccharomyces pombe. Its antibacterial activity was investigated on gram-negative bacteria Pseudomonas fluorescens. These activities were assessed by radial agar diffusion assay, and scanning and transmission electron microscopy. Therefore, destabilase-lysozyme not only is endo-isopeptidase and lysozyme, but also has antifungal and antibacterial activities. Muramidase activity of rec.Dest-Lys disappeared after heat-treating at 90°C for 50 min without the loss of its antimicrobial activity. Furthermore we showed for the first time that the heated-up derivative of rec.Dest-Lys exhibited more potent activities against the above enumerated fungi and gram-negative bacteria than original protein.

Keywords

recombinant destabilase-lysozyme (rec.Dest-Lys) / antimicrobial activity / antifungal activity / medicinal leech / heated-up derivative of rec.Dest-Lys (T-rec.Dest-Lys)

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Acknowledgments

This work was supported by the Russian Foundation for Basic Research (Grant 11-04-12048 ofi-m). The authors acknowledge V. N. Lazarev and S. A. Levitsky (Research Institute of Physicochemical Medicine, Moscow, Russia) for preparation of recombinant destabilase- lysozyme.

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2014 Higher Education Press and Springer-Verlag Berlin Heidelberg