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A neutral protease from Bacillus nematocida, another potential virulence factor in the infection against nematodes

机译：来自线虫芽孢杆菌的中性蛋白酶，是线虫感染的另一种潜在毒力因子

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摘要

A neutral protease (npr) (designated Bae16) toxic to nematodes was purified to homogeneity from the strain Bacillus nematocida. The purified protease showed a molecular mass of approximately 40 kDa and displayed optimal activity at 55 degrees C, pH 6.5. Bioassay experiments demonstrated that this purified protease could destroy the nematode cuticle and its hydrolytic substrates included gelatin and collagen. The gene encoding Bae16 was cloned, and the deduced amino acid sequence showed 94% sequence identity with npr gene from B. amyloliquefaciens, but had low similarity (13-43%) with the previously reported virulence serine proteases from fungi or bacteria, which reflected their differences. Recombinant mature Bae16 (rm-Bae16) was expressed in Escherichia coli BL21 using pET30 vector system, and its nematicidal activity confirmed that Bae16 could be involved in the infection process. Our present study revealed that the npr besides the known alkaline serine protease could serve as a potential virulence factor in the infection against nematodes, furthermore, the two proteases with different characteristics produced by the same strain co-ordinated efforts to kill nematodes. These data helped to understand the interaction between this bacterial pathogen and its host.
机译：对线虫有毒的中性蛋白酶（npr）（指定为Bae16）从线虫芽孢杆菌中纯化到同质。纯化的蛋白酶显示出约40 kDa的分子量，并在55摄氏度，pH 6.5下显示最佳活性。生物测定实验表明，这种纯化的蛋白酶可以破坏线虫的表皮，其水解底物包括明胶和胶原蛋白。克隆了编码Bae16的基因，推导的氨基酸序列与解淀粉芽孢杆菌的npr基因具有94％的序列同一性，但与以前报道的来自真菌或细菌的毒力丝氨酸蛋白酶的相似性较低（13-43％），这反映了他们的差异。使用pET30载体系统在大肠杆菌BL21中表达了重组成熟Bae16（rm-Bae16），其杀线虫活性证实Bae16可能参与了感染过程。我们目前的研究表明，除已知的碱性丝氨酸蛋白酶外，npr还可作为感染线虫的潜在毒力因子，此外，同一菌株产生的两种具有不同特性的蛋白酶协同作用来杀死线虫。这些数据有助于了解这种细菌病原体与其宿主之间的相互作用。

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