Molecular characterization and expression pattern of c-type and g-type lysozyme isoforms in starry flounder Platichthys stellate infected with Streptococcus parauberis

Kim, Yi Kyung; Nam, Yoon Kwon

doi:10.1007/s12562-015-0852-0

Cited by 13 publications

(3 citation statements)

References 57 publications

(77 reference statements)

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“…Several studies indicate the supplementation of chitin and its derivatives could elevate lysozyme activity in fish (Chen & Chen, 2019; Shanthi Mari et al., 2014; Taufek et al., 2018). In fish, lysozyme is expressed in the hematopoietic organs such as spleen, liver, kidney, gills and mucosal tissues (Kim & Nam, 2015; Saurabh & Sahoo, 2008). The lysozyme activity is dependent on the intensity of stress, infection conditions, and/or nutrition (Saurabh & Sahoo, 2008; Yildiz, 2006).…”

Section: Discussionmentioning

confidence: 99%

Nile tilapia fed insect meal: Growth and innate immune response in different times under lipopolysaccharide challenge

et al. 2020

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Section: Discussionmentioning

confidence: 99%

Nile tilapia fed insect meal: Growth and innate immune response in different times under lipopolysaccharide challenge

et al. 2020

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“…During its bactericidal activity, it biodegrades the bacterial cell wall by catalyzing the hydrolysis of the β-(1,4)-glycosidic bonds between N-acetylmuramic acid and N-acetylglucosamine in the peptidoglycan layer, which is the main component of the cell wall of both gram-positive and gram-negative bacteria [ 4 ]. Lysozyme has six major types, e.g., phage-lysozyme, bacterial-lysozyme, c(chicken)-type, g(goose)-type, I(invertebrate)-type, and plant lysozyme [ 5 ]. Bacterial lysozyme appears to be very similar to egg-white lysozyme [ 4 , 6 , 7 ].…”

Section: Introductionmentioning

confidence: 99%

Purification, Characterization and Bactericidal Action of Lysozyme, Isolated from Bacillus subtillis BSN314: A Disintegrating Effect of Lysozyme on Gram-Positive and Gram-Negative Bacteria

et al. 2023

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In the present study, lysozyme was purified by the following multi-step methodology: salt (ammonium sulfate) precipitation, dialysis, and ultrafiltration. The lysozyme potential was measured by enzymatic activity after each purification step. However, after ultrafiltration, the resulting material was considered extra purified. It was concentrated in an ultrafiltration centrifuge tube, and the resulting protein/lysozyme was used to determine its bactericidal potential against five bacterial strains, including three gram-positive (Bacillus subtilis 168, Micrococcus luteus, and Bacillus cereus) and two gram-negative (Salmonella typhimurium and Pseudomonas aeruginosa) strains. The results of ZOI and MIC/MBC showed that lysozyme had a higher antimicrobial activity against gram-positive than gram-negative bacterial strains. The results of the antibacterial activity of lysozyme were compared with those of ciprofloxacin (antibiotic). For this purpose, two indices were applied in the present study: antimicrobial index (AMI) and percent activity index (PAI). It was found that the purified lysozyme had a higher antibacterial activity against Bacillus cereus (AMI/PAI; 1.01/101) and Bacillus subtilis 168 (AMI/PAI; 1.03/103), compared to the antibiotic (ciprofloxacin) used in this study. Atomic force microscopy (AFM) was used to determine the bactericidal action of the lysozyme on the bacterial cell. The purified protein was further processed by gel column chromatography and the eluate was collected, its enzymatic activity was 21.93 U/mL, while the eluate was processed by native-PAGE. By this analysis, the un-denatured protein with enzymatic activity of 40.9 U/mL was obtained. This step shows that the protein (lysozyme) has an even higher enzymatic potential. To determine the specific peptides (in lysozyme) that may cause the bactericidal potential and cell lytic/enzymatic activity, the isolated protein (lysozyme) was further processed by the SDS-PAGE technique. SDS-PAGE analysis revealed different bands with sizes of 34 kDa, 24 kDa, and 10 kDa, respectively. To determine the chemical composition of the peptides, the bands (from SDS-PAGE) were cut, enzymatically digested, desalted, and analyzed by LC-MS (liquid chromatography-mass spectrometry). LC-MS analysis showed that the purified lysozyme had the following composition: the number of proteins in the sample was 56, the number of peptides was 124, and the number of PSMs (peptide spectrum matches) was 309. Among them, two peptides related to lysozyme and bactericidal activities were identified as: A0A1Q9G213 (N-acetylmuramoyl-L-alanine amidase) and A0A1Q9FRD3 (D-alanyl-D-alanine carboxypeptidase). The corresponding protein sequence and nucleic acid sequence were determined by comparison with the database.

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“…Among these lysozymes, C‐type lysozyme has the largest proportion in nature, and exists in vertebrates and insects. Till present, many kinds of C‐type lysozyme have been identified in different animals, such as Venerupis philippinarum (Yang et al, ), Trachinotus ovatus (Zhou et al, ), Harmonia axyridis (Beckert et al, ), Apostichopus japonicas (Tian, Liang, Chang, & Song, ), Carassius auratus (Wang et al, ) and Platichthys stellate (Kim & Nam, ), and its function has been studied. The C‐type lysozymes typically possess muramidase activities and chitinase activities, which could cleave the β‐(1,4)‐glycosidic bond of PGN and break down glycosidic bonds or N‐acetylglucosamine linkages of chitin (Fiołka, Ptaszyńska, & Czarniawski, ; Vocadlo, Davies, Laine, & Withers, ).…”

Section: Introductionmentioning

confidence: 99%

Newly identified C‐type lysozyme in Chinese soft‐shelled turtle (Pelodiscus sinensis) exhibits potent antimicrobial activity

Yuan

et al. 2019

Aquac Res

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Lysozymes play vital roles in humoural immune response against bacterial invasion by its lytic activity. In the present study, a new C‐type lysozyme was identified and characterized from Chinese soft‐shelled turtle Pelodiscus sinensis. The full‐length cDNA of PslysC was of 923 bp, encoding a polypeptide of 148 amino acid residues. The multiple alignments and phylogenetic relationship analysis revealed the highly enzyme‐related conserved residues. The real‐time PCR analysis suggested that PslysC was constitutively expressed in a wide range of tissues with highest level in blood cells and liver. The expression of PslysC could be significantly up‐regulated under Aeromonas jandaei infection and ammonia exposure, while no significant changes were found under Poly I:C infection. The rPslysC protein was expressed in E. coli and purified by Ni‐NTA. The optimal pH and temperature for rPslysC protein lytic activities were determined at pH 7 and 30℃. rPslysC can inhibit the growth of eight kinds of Gram‐negative bacteria, and three kinds of Gram‐positive bacteria. The binding activity of rPslysC to different microbial polysaccharides and microorganism was analysed. The results showed that rPslysC could bind to selected bacteria, and exhibit a strong binding activity to lipopolysaccharide and peptidoglycan, but a weak binding activity to β‐glucan. This suggests that the binding activity might be the major mechanism of action to realize the antibacterial activity. The present study will provide helpful evidence to further understand the innate immunity of P. sinensis, and the interaction mechanisms of C‐type lysozymes with bacterial membranes.

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Molecular characterization and expression pattern of c-type and g-type lysozyme isoforms in starry flounder Platichthys stellate infected with Streptococcus parauberis

Cited by 13 publications

References 57 publications

Nile tilapia fed insect meal: Growth and innate immune response in different times under lipopolysaccharide challenge

Nile tilapia fed insect meal: Growth and innate immune response in different times under lipopolysaccharide challenge

Purification, Characterization and Bactericidal Action of Lysozyme, Isolated from Bacillus subtillis BSN314: A Disintegrating Effect of Lysozyme on Gram-Positive and Gram-Negative Bacteria

Newly identified C‐type lysozyme in Chinese soft‐shelled turtle (Pelodiscus sinensis) exhibits potent antimicrobial activity

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