Fisheries and Aquatic Sciences

The Korean Society of Fisheries and Aquatic Science (한국수산과학회)
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Functional characterization and expression analysis of c-type and g-like-type lysozymes in yellowtail clownfish (Amphiprion clarkii)

  • Gaeun Kim (Department of Marine Life Sciences & Fish Vaccine Research Center, Jeju National University) ;
  • Hanchang Sohn (Department of Marine Life Sciences & Fish Vaccine Research Center, Jeju National University) ;
  • WKM Omeka (Department of Marine Life Sciences & Fish Vaccine Research Center, Jeju National University) ;
  • Chaehyeon Lim (Department of Marine Life Sciences & Fish Vaccine Research Center, Jeju National University) ;
  • Don Anushka Sandaruwan Elvitigala (Department of Basic Sciences and Social Science for Nursing, Faculty of Nursing, University of Colombo) ;
  • Jehee Lee (Department of Marine Life Sciences & Fish Vaccine Research Center, Jeju National University)
  • Received : 2022.06.09
  • Accepted : 2023.01.10
  • Published : 2023.03.31
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Abstract

Lysozymes are well-known antibacterial enzymes that mainly target the peptidoglycan layer of the bacterial cell wall. Animal lysozymes are mainly categorized as g-type, c-type, and i-type based on protein sequence and structural differences. In this study, c-type (AcLysC) and g-like-type (AcLysG-like) lysozymes from Amphiprion clarkii were characterized in silico via expressional and functional approaches. According to in silico analysis, open reading frames of AcLysC and AcLysG-like were 429 bp and 570 bp, respectively, encoding the corresponding polypeptide chains with 142 and 189 amino acids. Elevated expression levels of AcLysC and AcLysG-like were observed in the liver and the heart tissues, respectively, as evidenced by quantitative real-time polymerase chain reaction assays. AcLysC and AcLysG-like transcript levels were upregulated in gills, head kidney, and blood cells following experimental immune stimulation. Recombinant AcLysC exhibited potent lytic activity against Vibrio anguillarum, whereas recombinant AcLysG-like showed remarkable antibacterial activity against Vibrio harveyi and Streptococcus parauberis, which was further evidenced by scanning electron microscopic imaging of destructed bacterial cell walls. The findings of this study collectively suggest the potential roles of AcLysC and AcLysG-like in host immune defense.

Keywords

Acknowledgement

This research was supported by the Korea Institute of Marine Science & Technology Promotion (KIMST), funded by the Ministry of Oceans and Fisheries (20220570).

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