Purification and characterization of an antibacterial protein from dried fruiting bodies of the wild mushroom Clitocybe sinopica.

Abstract

A novel antibacterial protein with a molecular mass of 44 kDa has been isolated from dried fruiting bodies of the wild mushroom Clitocybe sinopica. Sodium dodecyl sulfate/polyacrylamide gel electrophoresis showed that the protein was composed of two subunits each with a molecular mass of 22 kDa. Its N-terminal amino-acid sequence, SVQATVNGDKML, has not been reported for other antimicrobial proteins. The purification protocol included ion exchange chromatography on DEAE-cellulose, CM-cellulose and Q-Sepharose, and gel filtration by fast protein liquid chromatography on Superdex 75. The antibacterial protein was adsorbed on all three ion exchangers. The antimicrobial activity profile of the protein against tested bacterial and fungal strains disclosed that it possessed potent antibacterial activity against Agrobacterium rhizogenes, A. tumefaciens, A. vitis, Xanthomonas oryzae and X. malvacearum with a minimum inhibitory concentration mostly below 0.6 microM. However, it had no antibacterial activity against Pseudomonas batatae, Erwinia herbicola, Escherichia coli, and Staphylococcus aureus, and no antifungal activity against Setosphaeria turcica, Fusarium oxysporum, Verticillium dahliae, Bipolaris maydis, and B. sativum. The antibacterial antivity against A. tumefaciens was stable after exposure to 20-60 degrees C for 30 min and to pH 4-9 for 1 h.

Evaluation of Different Protein Precipitates of a Wild Mushroom, Ganoderma Lucidum for Antibacterial Activities Against Human Pathogenic Bacteria

“…Later studies have shown that certain higher molecular weight peptides and proteins of mushroom species are also taking part in bioactivity. Various antifungal proteins from different fungi such as Clitocybe sinopica, Aspergillus giganteus, Aspergillus niger, Zygosaccharomyces bailii, and Tricholoma giganteum [19,[31][32][33][34] have been reported. An immune modulatory protein LZ-8 from G. lucidum was also proposed to have therapeutic effects on cancer and autoimmune disease [35].…”

“…Similarly, the bacterial growth inhibition at micromolar concentration of different mushroom AMPs has also been investigated earlier. For instance, the proteins purified from Clitocybe sinopica had potential antifungal activity toward Agrobacterium tumefaciens at 0.14-2.7 µM [19]. These proteins had also a broad spectrum of activities against a number of plant pathogenic bacteria including strains of Agrobacterium and Xanthomonas.…”

“…Mushrooms are supposed to be rich sources of various pharmaceutical compounds including AMPs [14][15][16][17][18][19][20]. Reishi mushroom (Ganoderma lucidum) is one of the most famous oriental mushrooms having a long history of medicinal use in China, Japan, and Asian countries.…”

Fungal proteinaceous compounds with multiple biological activities

“…Its hemolytic activity remained stable at pH 4.0-12.0, but activity was indiscernible at pH 2 and pH 13 and below pH 2 (Ngai and Ng 2006). A dimeric 44-kDa antibacterial hemolysin from mushroom Clitocybe sinopica demonstrated antibacterial activity against Xanthomonas malvacearum, Xanthomonas oryzae, Agrobacterium tumefaciens, A. rhizogenes, and A. vitis, but not against Escherichia coli, Erwinia herbicola, Pseudomonas batatae, and S. aureus, and no antifungal activity against Bipolaris maydis, B. sativum, Fusarium oxysporum, Setosphaeria turcica, and Verticillium dahlia (Zheng et al 2010).…”

Synthesis and Antibacterial Activities of Pleuromutilin Derivatives with Thiazole-5-Carboxamide and Thioether Moiety

“…The CH 2 Cl 2 layer was evaporated under reduced pressure and the resulting residue was chromatographed on silica gel (petroleum ester/ethyl acetate = 2:1) to give the target molecule; yields: 71-80%. The antibacterial activities of the target compounds in vitro were tested via an agar-well diffusion method 20,21 .…”