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首页> 外文期刊>Applied and Environmental Microbiology >Purification and Characterization of a High-Molecular-Weight Insecticidal Protein Complex Produced by the Entomopathogenic BacteriumPhotorhabdus luminescens
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Purification and Characterization of a High-Molecular-Weight Insecticidal Protein Complex Produced by the Entomopathogenic BacteriumPhotorhabdus luminescens

机译：昆虫病原细菌Photorhabdus luminescens产生的高分子量杀虫蛋白复合物的纯化和表征

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摘要

Photorhabdus luminescens is a gram-negative enteric bacterium that is found in association with entomopathogenic nematodes of the family Heterorhabditidae. The nematodes infect a variety of soil-dwelling insects. Upon entering an insect host, the nematode releases P. luminescens cells from its intestinal tract, and the bacteria quickly establish a lethal septicemia. When grown in peptone broth, in the absence of the nematodes, the bacteria produce a protein toxin complex that is lethal when fed to, or injected into the hemolymph of, Manduca sexta larvae and several other insect species. The toxin purified as a protein complex which has an estimated molecular weight of 1,000,000 and contains no protease, phospholipase, or hemolytic activity and only a trace of lipase activity. The purified toxin possesses insecticidal activity whether injected or given orally. Analyses of the denatured complex by sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed it to be composed of several protein subunits ranging in size from 30 to 200 kDa. The complex was further separated by native gel electrophoresis into three components, two of which retained insecticidal activity. The purified native toxin complex was found to be active in nanogram concentrations against insects representing four orders of the classInsecta.
机译：发光的光杆菌是一种革兰氏阴性肠细菌，发现它与异形螨科的致病性线虫有关。线虫感染多种居住在土壤中的昆虫。进入昆虫宿主后，线虫会从其肠道释放发光假单胞菌细胞，并且细菌会迅速建立致命的败血病。当在不存在线虫的情况下，在蛋白bro肉汤中生长时，细菌会产生一种蛋白质毒素复合物，将其喂食或注射到曼杜卡六倍体幼虫和其他几种昆虫的淋巴液中会致命。纯化为蛋白质复合物的毒素，估计分子量为1,000,000，不包含蛋白酶，磷脂酶或溶血活性，仅包含微量脂肪酶活性。纯化的毒素无论是注射还是口服都具有杀虫活性。通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳对变性的复合物进行分析，结果表明该复合物由几个蛋白亚基组成，大小从30至200 kDa不等。通过天然凝胶电泳将该复合物进一步分离成三个组分，其中两个保留了杀虫活性。发现纯化的天然毒素复合物在纳克浓度下对代表class Insecta四个阶的昆虫具有活性。

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