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Mycopathologia

, Volume 163, Issue 3, pp 169–176 | Cite as

Purification and characterization of a neutral serine protease with nematicidal activity from Hirsutella rhossiliensis

  • Bin Wang
  • Wenping Wu
  • Xingzhong LiuEmail author
Article

Abstract

Serine protease plays an important role in fungal infection to invertebrate hosts. An extracellular protease (Hnsp) was detected in liquid culture of Hirsutella rhossiliensis OWVT-1 with nematodes (Panagrellus redivivus) as the unique nitrogen source and purified to homogeneity by ammonium sulphate precipitation, anion exchange chromatography and gel filtration. Its molecular mass was about 32 kDa, and the optimal reaction pH value and temperature were pH 7 and 40°C, respectively. The Hnsp activity was stable at pH 6–8 and decreased radically at 50°C for 10 min. Hnsp was highly sensitive to inhibitor of PMSF and well decomposed the substrate N-succinyl-Ala-Ala-Pro-Phe-p-nitroanilide, suggesting that it belonged to the chymotrypsin/subtilisin of serine proteases. The N-terminal amino acid sequence of Hnsp was SVTDQQGADCGLARISHRE, which showed high homology with other serine proteases from nematophagous fungi. Ability to kill nematode and degrade its cuticle in vitro indicated that Hnsp could be involved in the infection of nematode.

Keywords

characterization Hirsutella rhossiliensis nematophagous fungus purification serine protease 

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Copyright information

© Springer 2007

Authors and Affiliations

  1. 1.Key Laboratory of Systematic Mycology and LichenologyInstitute of Microbiology, Chinese Academy of SciencesBeijingChina
  2. 2.Graduate School of Chinese Academy of SciencesBeijingChina
  3. 3.Novozymes ChinaHaidian District, BeijingChina

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