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出 处:《水生生物学报》2004年第4期409-412,共4页Acta Hydrobiologica Sinica
基 金:中国科学院知识创新工程资助项目 (KSCX2 1 0 4);"十五"国家高技术研究发展计划 ( 863计划 )资助项目 ( 2 0 0 3AA62 2 0 5 0 );广东省"科技创新百项工程"资助项目 ( 99B0 62 0 1G)
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 4 5 6kD ;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 2 6 6 % ,碱性氨基酸含量为 14 4 % ,疏水性氨基酸含量为 33 4 % ;毒素活性的最适pH为 6— 8,对胰酶有抗性 ;不耐热 ,10 0℃下作用 1min ,其毒性完全消失 ,而在 5 6℃下 ,毒素完全失去活性需要 4 0min。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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