海水养殖真鲷弧菌病病原菌外毒素的理化特性

THE CHARACTERISTICS OF THE EXOTOXIN Vm-Pm PRODUCED BY VIBRIO MIMICUS

  • 摘要: 对最小弧菌Vm-Pm毒素的分子量、分子结构及氨基酸组分进行了测定,同时还研究了温度、酸碱度及胰酶等因子对该毒素生物学活性的影响.研究结果表明,该毒素为单一的多肽,不具备典型的细菌毒素A、B亚基结构,分子量为30.56kD;毒素蛋白含天冬氨酸、苏氨酸、丝氨酸及半胱氨酸等15种氨基酸,其中天冬氨酸的含量最多,半胱氨酸的含量最少,酸性氨基酸含量为26.6%,碱性氨基酸含量为1.%,疏水性氨基酸含量为33.%;毒素活性的最适pH为6-8,对胰酶有抗性;不耐热,100℃下作用1min,其毒性完全消失,而在56℃下,毒素完全失去活性需要40min.

     

    Abstract: Toxin Vm-Pm produced by Vibrio mimicus was purified in procedures as follows:salting out of ammonium sulfate,anion exchange chromatography on DEAE-cellulose and gel filtration chromatography of Sephadex G100.The purified toxin Vm-Pm gave a single band with a molecular weight of 30.456kD following SDS-PAGE with or without β-mercaptoethanol,indicating that the toxin Vm-Pm did not contain inter-chain disulphide bonds in its conformation and was a single-chain polypeptide.Amino acid composition analysis demonstrated that the toxin protein contained fifteen different kinds of amino acids and revealed a comparatively higher percentage of acidic amino acids(26.6%)than alkalic amino acids(14.4%).Hydrophobic amino acids corresponded to 33.4% of total amino acid residues.The optimal pH for the toxin was 6-8 and resistant to trypsase. Heat treatment at 56℃ and 100℃ abolished the hemolytic activity of the toxin.The toxin completely lost its activity when incubated at temperatures of 56℃ and 100℃ for 40min and 1min,respectively. The toxin showed unique characteristics as compared with other known bacterial toxins.

     

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