Identification and Functional Analysis of a Lysozyme Gene from Coridius chinensis (Hemiptera: Dinidoridae)

SIMPLE SUMMARY: As a medicinal insect, Coridius chinensis contains many active polypeptides. Extracts from C. chinensis are usually complex and it is not clear which polypeptides are effective medicinal ingredients. In addition, we also need to figure out the functions of various immune effectors in the innate immunity of C. chinensis. To explore the function of lysozyme in C. chinensis, a lysozyme gene CcLys2 was screened and identified from the transcriptome data of C. chinensis. The results showed that CcLys2 had a typical domain of the c-type lysozyme, belonging to the H-branch of the c-type lysozyme. The lysozyme Cclys2 is an effective immune effector in the immune response of C. chinensis and can be stimulated by bacterial infection. Like typical c-type lysozyme, Cclys2 has lytic activity against Gram-positive bacteria. The research holds promise for functional annotation of similar proteins from other dinidoridae insects and provides the theoretical feasibility for the development of medicinal components in C. chinensis. Our results also provide data for further investigating the origin and evolution of insect lysozymes. ABSTRACT: Coridius chinensis is a valuable medicinal insect resource in China. Previous studies have indicated that the antibacterial and anticancer effects of the C. chinensis extract mainly come from the active polypeptides. Lysozyme is an effective immune effector in insect innate immunity and usually has excellent bactericidal effects. There are two kinds of lysozymes in insects, c-type and i-type, which play an important role in innate immunity and intestinal digestion. Studying lysozyme in C. chinensis will be helpful to further explore the evolutionary relationship and functional differences among lysozymes of various species and to determine whether they have biological activity and medicinal value. In this study, a lysozyme CcLys2 was identified from C. chinensis. CcLys2 contains 223 amino acid residues, and possesses a typical domain of the c-type lysozyme and a putative catalytic site formed by two conserved residues Glu32 and Asp50. Phylogenetic analysis showed that CcLys2 belongs to the H-branch of the c-type lysozyme. The analysis of spatiotemporal expression patterns indicated that CcLys2 was mainly expressed in the fat body of C. chinensis adults and was highly expressed in the second- and fifth-instar nymphs. In addition, CcLys2 was significantly up-regulated after injecting and feeding bacteria. In the bacterial inhibition assay, it was found that CcLys2 had antibacterial activity against Gram-positive bacteria at a low pH. These results indicate that CcLys2 has muramidase activity, involves in the innate immunity of C. chinensis, and is also closely related to the bacterial immune defense or digestive function of the intestine.