Purification and characterization of an antibacterial protein from dried fruiting bodies of the wild mushroom Clitocybe sinopica.
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.
Zheng
Suyue
S
State Key Laboratory for Agrobiotechnology and Department of Microbiology, China Agricultural University, Beijing, China.
Liu
Qinghong
Q
Zhang
Guoqing
G
Wang
Hexiang
H
Ng
Tzi Bun
TB
D000363:Agaricales / Q000737:chemistry*; D016960:Agrobacterium tumefaciens / Q000187:drug effects / Q000254:growth & development; D000595:Amino Acid Sequence; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D005656:Fungal Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology*; D006863:Hydrogen-Ion Concentration; D050296:Microbial Viability / Q000187:drug effects; D008969:Molecular Sequence Data; D055550:Protein Stability
D005656:Fungal Proteins