title: Identification and characterization of a novel phage-type like lysozyme from Manila clam, Ruditapes philippinarum.
abstract: A novel lysozyme gene (RpLysPh) with high similarity to the bacteriophage lysozymes was identified in Manila clam, Ruditapes philippinarum. The full length cDNA of RpLysPh is 828bp and contains a 462bp open reading frame (ORF) that codes for a 154 amino acid protein. Multiple sequence alignment analysis revealed that the three residues essential for catalytic activity in phage-type lysozyme (Glu(20), Asp(29), and Thr(35)) are conserved in RpLysPh. The comparison of the 3D models of RpLysPh and Coxiella burnetii lysozyme also suggested that the active sites involved in the binding of substrate have similar conformations. Phylogenetic analysis suggested that RpLysPh shares a similar origin with the bacterial phage-type lysozyme group. The highest level of expression of RpLysPh was observed in hemocytes, followed by mantle. Induction of RpLysPh expression was observed in gills in response to lipopolysaccharide (LPS), peptidoglycan (PGN), polyinosinic-polycytidylic acid (Poly(I:C)), and whole glucan particles (WGP) challenge. The recombinant protein of RpLysPh showed antibacterial activity against both Gram-positive and Gram-negative bacteria.
authors.affiliation: Dalian Ocean University, Dalian 116023, PR China; Engineering Research Center of Shellfish Culture and Breeding in Liaoning Province, Dalian 116023, PR China.
authors.forename: Jianfeng
authors.initials: J
authors.lastname: Ding
authors.affiliation: Dalian Ocean University, Dalian 116023, PR China; Engineering Research Center of Shellfish Culture and Breeding in Liaoning Province, Dalian 116023, PR China.
authors.forename: Rui
authors.initials: R
authors.lastname: Wang
authors.affiliation: Dalian Ocean University, Dalian 116023, PR China; Engineering Research Center of Shellfish Culture and Breeding in Liaoning Province, Dalian 116023, PR China.
authors.forename: Feng
authors.initials: F
authors.lastname: Yang
authors.affiliation: Dalian Ocean University, Dalian 116023, PR China; Engineering Research Center of Shellfish Culture and Breeding in Liaoning Province, Dalian 116023, PR China.
authors.forename: Liqiang
authors.initials: L
authors.lastname: Zhao
authors.affiliation: Dalian Ocean University, Dalian 116023, PR China; Engineering Research Center of Shellfish Culture and Breeding in Liaoning Province, Dalian 116023, PR China.
authors.forename: Yanjie
authors.initials: Y
authors.lastname: Qin
authors.affiliation: Institute of Oceanology, Chinese Academy of Sciences, Qingdao 266071, PR China.
authors.forename: Guofan
authors.initials: G
authors.lastname: Zhang
authors.affiliation: Dalian Ocean University, Dalian 116023, PR China; Engineering Research Center of Shellfish Culture and Breeding in Liaoning Province, Dalian 116023, PR China. Electronic address: yanxiwu@dlou.edu.cn.
authors.forename: Xiwu
authors.initials: X
authors.lastname: Yan
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D049872:Bivalvia / Q000201:enzymology*; D006434:Hemocytes / Q000201:enzymology; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009113:Muramidase / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D010802:Phylogeny; D011994:Recombinant Proteins / Q000302:isolation & purification / Q000378:metabolism; D016415:Sequence Alignment; D059467:Transcriptome
chemical_list: D011994:Recombinant Proteins; D009113:Muramidase
keywords: Antibacterial activity; Basal and temporal transcriptional analysis; Phage-type like lysozyme; Ruditapes philippinarum