title: The recombinant serine protease XAoz1 of Arthrobotrys oligospora exhibits potent nematicidal activity against Caenorhabditis elegans and Haemonchus contortus.
abstract: The nematophagous fungus Arthrobotrys oligospora is a potential biological agent against parasitic gastrointestinal nematodes. Its subtilisin-like serine proteases play an important role in nematode cuticle breach. In this study, the cDNA of the mature serine protease XAoz1 from A. oligospora XJ-XAo1 was expressed in Pichia pastoris to assess the in vitro nematicidal activity of recombinant XAoz1 (reXAoz1) on Caenorhabditis elegans and Haemonchus contortus. The cDNA sequence of the protease XAoz1 was amplified by reverse transcription polymerase chain reaction (RT-PCR) and inserted into the vector pPIC9K for expression in P.pastoris GS115. Our results show that the reXAoz1 had a molecular mass of 50 kDa after 3 days of 1.5%-methanol induction at 28 °C. The highest specific protease activity was achieved at 12 168 U mg(-1) protein. The reXAoz1 had the highest hydrolytic activity at pH 6.5-9.5 with an optimal pH at 8.5. Moreover, the purified reXAoz1 displayed a highly toxic and biological activity to immobilize C. elegans and H. contortus by degrading their cuticles and inducing death.
authors.affiliation: Key Lab of Preventive Veterinary, College of Animal Science and Technology, Shihezi University, Shihezi, Xinjiang, China.
authors.forename: Wang
authors.initials: W
authors.lastname: Junwei
authors.forename: Meng
authors.initials: M
authors.lastname: Qingling
authors.forename: Qiao
authors.initials: Q
authors.lastname: Jun
authors.forename: Wang
authors.initials: W
authors.lastname: Weisheng
authors.forename: Chen
authors.initials: C
authors.lastname: Shuangqing
authors.forename: Luo
authors.initials: L
authors.lastname: Jianxun
authors.forename: Zhao
authors.initials: Z
authors.lastname: Chunguang
authors.forename: Chen
authors.initials: C
authors.lastname: Chuangfu
mesh_terms: D000818:Animals; D000969:Antinematodal Agents / Q000494:pharmacology*; D001203:Ascomycota / Q000201:enzymology* / Q000235:genetics; D017173:Caenorhabditis elegans / Q000187:drug effects*; D003001:Cloning, Molecular; D004789:Enzyme Activation; D015870:Gene Expression; D006189:Haemonchus / Q000187:drug effects*; D006863:Hydrogen-Ion Concentration; D011994:Recombinant Proteins / Q000494:pharmacology*; D057057:Serine Proteases / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism / Q000494:pharmacology*
chemical_list: D000969:Antinematodal Agents; D011994:Recombinant Proteins; D057057:Serine Proteases