Molecular characterization, expression and antimicrobial activities of two c-type lysozymes from manila clam Venerupis philippinarum

Dinglong Yang; Qing Wang; Ruiwen Cao; Lizhu Chen; Yongliang Liu; Ming Cong; Huifeng Wu; Fei Li; Chenglong Ji; Jianmin Zhao

doi:10.1016/j.dci.2017.03.018

Molecular characterization, expression and antimicrobial activities of two c-type lysozymes from manila clam Venerupis philippinarum

Dev Comp Immunol. 2017 Aug:73:109-118. doi: 10.1016/j.dci.2017.03.018. Epub 2017 Mar 24.

Authors

Dinglong Yang¹, Qing Wang², Ruiwen Cao³, Lizhu Chen³, Yongliang Liu⁴, Ming Cong², Huifeng Wu², Fei Li², Chenglong Ji², Jianmin Zhao⁵

Affiliations

¹ Key Laboratory of Coastal Zone Environmental Processes, Yantai Institute of Coastal Zone Research (YIC), Chinese Academy of Sciences (CAS), Yantai, Shandong 264003, PR China; Muping Coastal Environment Research Station, Yantai Institute of Coastal Zone Research, Chinese Academy of Sciences, Yantai 264003, PR China; University of Chinese Academy of Sciences, Beijing 100049, PR China.
² Key Laboratory of Coastal Zone Environmental Processes, Yantai Institute of Coastal Zone Research (YIC), Chinese Academy of Sciences (CAS), Yantai, Shandong 264003, PR China.
³ Key Laboratory of Coastal Zone Environmental Processes, Yantai Institute of Coastal Zone Research (YIC), Chinese Academy of Sciences (CAS), Yantai, Shandong 264003, PR China; University of Chinese Academy of Sciences, Beijing 100049, PR China.
⁴ Muping Coastal Environment Research Station, Yantai Institute of Coastal Zone Research, Chinese Academy of Sciences, Yantai 264003, PR China.
⁵ Key Laboratory of Coastal Zone Environmental Processes, Yantai Institute of Coastal Zone Research (YIC), Chinese Academy of Sciences (CAS), Yantai, Shandong 264003, PR China; Muping Coastal Environment Research Station, Yantai Institute of Coastal Zone Research, Chinese Academy of Sciences, Yantai 264003, PR China. Electronic address: jmzhao@yic.ac.cn.

PMID: 28344168
DOI: 10.1016/j.dci.2017.03.018

Abstract

Lysozymes play an important role in the innate immune responses with which mollusks respond to bacterial invasion through its lytic activity. In the present study, two c-type lysozymes (designed as VpCLYZ-1 and VpCLYZ-2, respectively) were identified and characterized from the manila clam Venerupis philippinarum. The full-length cDNA of VpCLYZ-1 and VpCLYZ-2 was of 629 and 736 bp, encoding a polypeptide of 156 and153 amino acid residues, respectively. The deduced amino acid sequences of VpCLYZs showed high similarity to other known invertebrate c-type lysozymes. Multiple alignments and phylogenetic relationship strongly suggested that VpCLYZ-1 and VpCLYZ-2 belonged to the c-type lysozyme family. Both VpCLYZ-1 and VpCLYZ-2 transcripts were constitutively expressed in a wide range of tissues with different levels. The VpCLYZ-1 transcript was dominantly expressed in hepatopancreas and hemocytes, while VpCLYZ-2 transcript was mainly expressed in the tissues of hepatopancreas and gills. Both the mRNA expression of VpCLYZ-1 and VpCLYZ-2 was significantly up-regulated at 12 h post Vibrio anguillarum challenge. The recombinant VpCLYZ-1 and VpCLYZ-2 (designed as rVpCLYZ-1 and rVpCLYZ-2) exhibited lytic activity against all tested bacteria, and rVpCLYZ-1 showed higher activities than rVpCLYZ-2 in killing Micrococcus luteus and V. anguillarum. Overall, our results suggested that VpCLYZ-1 and VpCLYZ-2 belonged to the c-type lysozyme family, and played important roles in the immune responses of manila clam, especially in the elimination of pathogens.

Keywords: Antimicrobial activity; C-type lysozyme; Muramidase activity; Venerupis philippinarum.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Bivalvia / enzymology
Bivalvia / immunology*
Gene Expression Profiling
Immunity, Innate / immunology*
Muramidase / biosynthesis
Muramidase / immunology*
Transcriptome

Substances

Muramidase