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
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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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Keywords:
- Pagrus major /
- Vibriosis /
- Vibrio mimicus /
- Vm-Pm toxin
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[1] Wu H B,Pan J P.The immune protective effects of three kinds of vaccination against vibriosis in marine-cage cultured red sea bream(Pagrus major)[J].Acta Hydrobiol.Sinica,2002,26 (5):457-464[吴后波,潘金培.三种免疫制剂对真鲷弧菌病的免疫保护性.水生生物学报,2002,26(5):457-464][2] Wu H B,Pan J P.Purification and characterization of the exotoxin Vm-Pm produced by Vibrio mimicus[J].Oceanologia et Limnologia sinica,2002,33(1):83-89[吴后波,潘金培.海水养殖真鲷弧菌病病原菌外毒素的分离、纯化及生物学活性.海洋与湖沼,2002,33(1):83-89][3] Wu H B,Pan J P.Effects of cultural conditions on the production of toxin by Vibrio mimicus[J].Acta Hydrobiol.Sinica,2003,27(1):30-32[吴后波,潘金培.培养条件对养殖真鲷弧菌病病原菌毒素产生的影响.水生生物学报,2003,27(1):30-32][4] Yoshida H,Honda T,Miwatani T.Purification and characterization of a hemolysin of Vibrio mimicus that relates to thermostable direct hemolysin of Vibrio parahaemolyticus[J].FEMS Microbiol.Lett.,1991,84:249-254[5] Miyoshi S, Sasahara K, Akamatsu S, et al.Purification and characterization of a hemolysin produced by Vibrio mimicus[J].Infect.Immun.1997,65 :1830-1835[6] Spira W M,Fedorka-Cray P J.Purification of enterotoxins from Vibrio mimicus that appear to be identical to cholera toxin[J].Infect.Immun.,1984,45 :679-684[7] Chowdhury M A R,Aziz K M S,Kay B A,et al.Toxin produced by Vibrio mimicus strains isolated from human and environmental sources in Bangladesh[J].J.Clin.Microbiol.,1987,25:2200-2203[8] Gyobu Y,Kodama H,Uetake H.Production and partial purification of a fluid-accumulating factor of non-01 Vibrio cholerae[J].Microbiol.Immunol.,1988,32 :565-577 Wu H B,Pan J P.The immune protective effects of three kinds of vaccination against vibriosis in marine-cage cultured red sea bream(Pagrus major)[J].Acta Hydrobiol.Sinica,2002,26 (5):457-464[吴后波,潘金培.三种免疫制剂对真鲷弧菌病的免疫保护性.水生生物学报,2002,26(5):457-464][2] Wu H B,Pan J P.Purification and characterization of the exotoxin Vm-Pm produced by Vibrio mimicus[J].Oceanologia et Limnologia sinica,2002,33(1):83-89[吴后波,潘金培.海水养殖真鲷弧菌病病原菌外毒素的分离、纯化及生物学活性.海洋与湖沼,2002,33(1):83-89][3] Wu H B,Pan J P.Effects of cultural conditions on the production of toxin by Vibrio mimicus[J].Acta Hydrobiol.Sinica,2003,27(1):30-32[吴后波,潘金培.培养条件对养殖真鲷弧菌病病原菌毒素产生的影响.水生生物学报,2003,27(1):30-32][4] Yoshida H,Honda T,Miwatani T.Purification and characterization of a hemolysin of Vibrio mimicus that relates to thermostable direct hemolysin of Vibrio parahaemolyticus[J].FEMS Microbiol.Lett.,1991,84:249-254[5] Miyoshi S, Sasahara K, Akamatsu S, et al.Purification and characterization of a hemolysin produced by Vibrio mimicus[J].Infect.Immun.1997,65 :1830-1835[6] Spira W M,Fedorka-Cray P J.Purification of enterotoxins from Vibrio mimicus that appear to be identical to cholera toxin[J].Infect.Immun.,1984,45 :679-684[7] Chowdhury M A R,Aziz K M S,Kay B A,et al.Toxin produced by Vibrio mimicus strains isolated from human and environmental sources in Bangladesh[J].J.Clin.Microbiol.,1987,25:2200-2203[8] Gyobu Y,Kodama H,Uetake H.Production and partial purification of a fluid-accumulating factor of non-01 Vibrio cholerae[J].Microbiol.Immunol.,1988,32 :565-577
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