Parasites, Hosts and Diseases

The Korea Society for Parasitology and Tropical Medicine (대한기생충학·열대의학회)
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Biological Characteristics of Recombinant Arthrobotrys oligospora Chitinase AO-801

  • Gong, Shasha (College of Animal Science & Technology, Shihezi University) ;
  • Meng, Qingling (College of Animal Science & Technology, Shihezi University) ;
  • Qiao, Jun (College of Animal Science & Technology, Shihezi University) ;
  • Huang, Yunfu (College of Animal Science & Technology, Shihezi University) ;
  • Zhong, Wenqiang (College of Animal Science & Technology, Shihezi University) ;
  • Zhang, Guowu (College of Animal Science & Technology, Shihezi University) ;
  • Zhang, Kai (College of Animal Science & Technology, Shihezi University) ;
  • Li, Ningxing (College of Animal Science & Technology, Shihezi University) ;
  • Shang, Yunxia (College of Animal Science & Technology, Shihezi University) ;
  • Li, Zhiyuan (College of Animal Science & Technology, Shihezi University) ;
  • Cai, Xuepeng (State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences)
  • Received : 2022.08.06
  • Accepted : 2022.09.15
  • Published : 2022.10.31
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Abstract

Chitinase AO-801 is a hydrolase secreted by Arthrobotrys oligospora during nematode feeding, while its role remained elusive. This study analyzed the molecular characteristics of recombinant chitinase of Arthrobotrys oligospora (reAO-801). AO-801 belongs to the typical glycoside hydrolase 18 family with conserved chitinase sequence and tertiary structure of (α/β)8 triose-phosphate isomerase (TIM) barrel. The molecular weight of reAO-801 was 42 kDa. reAO-801 effectively degraded colloidal and powdered chitin, egg lysate, and stage I larval lysate of Caenorhabditis elegans. The activity of reAO-801 reached its peak at 40℃ and pH values between 4-7. Enzyme activity was inhibited by Zn2+, Ca2+, and Fe3+, whereas Mg2+ and K+ potentiated its activity. In addition, urea, sodium dodecyl sulfate, and 2-mercaptoethanol significantly inhibited enzyme activity. reAO-801 showed complete nematicidal activity against C. elegans stage I larvae. reAO-801 broke down the C. elegans egg shells, causing them to die or die prematurely by hatching the eggs. It also invoked degradation of Haemonchus contortus eggs, resulting in apparent changes in the morphological structure. This study demonstrated the cytotoxic effect of reAO-801, which laid the foundation for further dissecting the mechanism of nematode infestation by A. oligospora.

Keywords

Acknowledgement

This work was supported by the National Natural Science Foundation of China (32060801). We thank the staff who provided the technical assistance for this study.

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