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

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Abstract

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°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.

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Acknowledgements

We are grateful to W. Zhou for her invaluable help in facilitating the work, H. Luo, J. Xu, B.Y. Tian, H. Sun, L.Q. Dong, J.W. Liu and L.H. Lian for their helpful advice in the study. This work was supported by the projects from National Natural Science Foundation Program of People’s Republic of China (30500338), Department of Science and Technology of Yunnan Province, People’s Republic of China (nos. 2005NG05, 2004C0004Q and 2004C0001Z).

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Authors and Affiliations

  1. Laboratory for Conservation and Utilization of Bio-resources, Yunnan University, Kunming, 650091, People’s Republic of China

    Qiuhong Niu, Xiaowei Huang, Lin Zhang, Yunxia Li, Juan Li, Jinkui Yang & Keqin Zhang

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  1. Qiuhong Niu
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  2. Xiaowei Huang
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  3. Lin Zhang
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  4. Yunxia Li
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  5. Juan Li
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  7. Keqin Zhang
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Correspondence to Keqin Zhang.

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Qiuhong Niu, Xiaowei Huang have contributed equally to this work.

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Niu, Q., Huang, X., Zhang, L. et al. A neutral protease from Bacillus nematocida, another potential virulence factor in the infection against nematodes. Arch Microbiol 185, 439–448 (2006). https://doi.org/10.1007/s00203-006-0112-x

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  • DOI: https://doi.org/10.1007/s00203-006-0112-x

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