Functional characterization and expression analysis of c-type and g-like-type lysozymes in yellowtail clownfish (Amphiprion clarkii)

Abstract

Lysozymes are well-known antibacterial enzymes that mainly target the peptidoglycan layer of the bacterial cell wall. Animal ly-sozymes 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.