Construction of ‘‘Toxin Complex’’ in a Mutant Serotype C Strain of Clostridium botulinum Harboring a Defective Neurotoxin Gene
Construction of ‘‘Toxin Complex’’ in a Mutant Serotype C Strain of Clostridium botulinum Harboring a Defective Neurotoxin Gene
复制标题
在带有缺陷神经毒素基因的肉毒梭菌 C 血清型突变菌株中构建“毒素复合物”
DOI：
10.1007/s00284-016-1150-5
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发表时间：
2016
影响因子：
2.6
通讯作者：
and Toshihiro Watanabe
中科院分区：
文献类型：
--
作者：
Tomonori Suzuki;Thomas Nagano;Koichi Niwa;Masataka Uchino;Motohiro Tomizawa;Yoshimasa Sagane;and Toshihiro Watanabe
一种产毒血清型斯德哥尔摩肉毒杆菌（C-St）的非产毒突变体C-N71不产生肉毒神经毒素（BoNT）。然而，原始菌株C-St产生肉毒杆菌毒素复合物，其中BoNT与无毒的非血凝素（NTNHA）和三种血凝素蛋白（HA-70， HA-33和HA-17）相关。因此，在本研究中，我们旨在阐明敲除bontgene对“毒素复合物”形成的影响。核苷酸序列分析表明，该基因中引入了一个过早终止密码子，而其他基因不受该突变的影响。此外，我们成功地纯化了C-N71产生的“毒素复合物”。“毒素复合物”被鉴定为NTNHA/HA-70/HA-17/HA-33复合物与完整的NTNHA或c端截断的NTNHA的混合物，没有BoNT。这些结果表明，敲除bontgene并不影响“毒素复合物”的形成。由于肉毒杆菌毒素复合物已被证明在人类和动物体内的口服毒素运输中起重要作用，因此C-N71的非神经毒性“毒素复合物”可能对口服药物递送系统的开发有价值。
A non-toxigenic mutant of the toxigenic serotype CClostridium botulinumstrain Stockholm (C-St), C-N71, does not produce the botulinum neurotoxin (BoNT). However, the original strain C-St produces botulinum toxin complex, in which BoNT is associated with non-toxic non-hemagglutinin (NTNHA) and three hemagglutinin proteins (HA-70, HA-33, and HA-17). Therefore, in this study, we aimed to elucidate the effects ofbontgene knockout on the formation of the “toxin complex.” Nucleotide sequence analysis revealed that a premature stop codon was introduced in thebontgene, whereas other genes were not affected by this mutation. Moreover, we successfully purified the “toxin complex” produced by C-N71. The “toxin complex” was identified as a mixture of NTNHA/HA-70/HA-17/HA-33 complexes with intact NTNHA or C-terminally truncated NTNHA, without BoNT. These results indicated that knockout of thebontgene does not affect the formation of the “toxin complex.” Since the botulinum toxin complex has been shown to play an important role in oral toxin transport in the human and animal body, a non-neurotoxic “toxin complex” of C-N71 may be valuable for the development of an oral drug delivery system.