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

doi:10.1007/s11705-012-1277-2

Front Chem Sci Eng

2012, Vol. 6

Issue (2) : 203-209 https://doi.org/10.1007/s11705-012-1277-2

RESEARCH ARTICLE

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

T. G. YUDINA¹, Danyang GUO¹(

), N. F. PISKUNKOVA¹, I. B. PAVLOVA¹, L. L. ZAVALOVA², I. P. BASKOVA¹

1. Biological Faculty, M. V. Lomonosov Moscow State University, Moscow 119991, Russia; 2. Institute of Bioorganic Chemistry (RAS) Miklukho-Maklaya 16/10, Moscow 117992, Russia

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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)

Corresponding Author(s): GUO Danyang,Email:guody1982@gmail.com

Issue Date: 05 June 2012

Cite this article:

T. G. YUDINA,Danyang GUO,N. F. PISKUNKOVA, et al. Antifungal and antibacterial functions of medicinal leech recombinant destabilase-lysozyme and its heated-up derivative[J]. Front Chem Sci Eng, 2012, 6(2): 203-209.

URL:

https://academic.hep.com.cn/fcse/EN/10.1007/s11705-012-1277-2
https://academic.hep.com.cn/fcse/EN/Y2012/V6/I2/203

Fig.1 Light microscopy of cells ( × 400) after their incubation with rec.Dest-Lys and T-rec.Dest-Lys (20 μg/mL) at 30°C for 3 h.
(a) Control; (b) lysis of fungal hyphae caused by rec-Dest-Lys; (c) more distinct lysis under the action of T-rec.Dest-Lys (experimental conditions were identical for both rec.Dest-Lys forms)

Fig.2 TEM of lysing hyphaes after their incubation with T-rec.Dest-Lys.
(a) Reduction of cytoplasm volume, formation of circular membrane structures; (b) degradation of cell wall and membrane segments; (c) cross-section of mycelium; disruption of cell walls, formation of circular membrane structures and lysis of their residues are seen

Fig.3 SEM of lyzed by rec.Dest-Lys and T-rec.Dest-Lys (20 μg/mL) at 30°C for 3 h.
(a) Control; (b) lysis of fungal hyphae caused by rec-Dest-Lys; (c) more distinct lysis under the action of T-rec.Dest-Lys (experimental conditions were identical for both rec.Dest-Lys forms)

Tab.1 Percentage of lysing cells of two fungi by rec.Dest-Lys and T-rec.Dest-Lys

Fig.4 TEM of cross-sections of lyzed by T-rec.Dest-Lys.
(a) Control; (b) cell wall loosening and start of stratification of CW, cytoplasm clarification and start of membrane backing up; (c) deterioration of CW outer layers, their lysis (more intense in young budding cells), and loss of a considerable part of inner cellular components; (d) swollen due to osmotic lysis cell (autophagy); (e) stratifications of capsule, its local degradation and formation of apoptosis structures

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