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Biochemical characterization of LysVpKK5 endolysin from a marine vibriophage.
- Source :
-
Protein expression and purification [Protein Expr Purif] 2021 Dec; Vol. 188, pp. 105971. Date of Electronic Publication: 2021 Sep 09. - Publication Year :
- 2021
-
Abstract
- Endolysins have been proposed as a potential antibacterial alternative for aquaculture, especially against Vibrio; the bacterial-agents that most frequently cause disease. Although multiple marine vibriophages have been characterized to date, research on vibriophage endolysins is recent. In this study, biochemical characterization of LysVpKK5 endolysin encoded by Vibrio parahaemolyticus-infecting VpKK5 phage was performed. In silico analysis revealed that LysVpKK5 possesses a conserved amidase&#95;2 domain with a zinc-binding motif of high structural similarity to T7 lysozyme (RMSD = 0.107 Å). Contrary to expectations, the activity was inhibited with Zn <superscript>2+</superscript> and was improved with other divalent cations, especially Ca <superscript>2+</superscript> . It showed optimal muralytic activity at pH 10, and curiously, no lytic activity at pH ≤ 7 was recorded. As for the thermal stability test, the optimal activity was recorded at 30 °C; the higher residual activity was recorded at 4 °C, and was lost at ≥ 50 °C. On the other hand, increasing NaCl concentrations reduced the activity gradually; the optimal activity was recorded at 50 mM NaCl. On the other hand, the enzymatic activity at 0.5 M NaCl was approx 30% and of approx 50% in seawater. LysVpKK5 endolysin exhibited a higher activity on V. parahaemolyticus ATCC-17802 strain, in comparison with AHPND + strains.<br /> (Copyright © 2021 Elsevier Inc. All rights reserved.)
- Subjects :
- Amino Acid Sequence
Aquatic Organisms
Bacteriophages classification
Bacteriophages genetics
Bacteriophages metabolism
Binding Sites
Calcium chemistry
Calcium pharmacology
Cations, Divalent
Endopeptidases chemistry
Endopeptidases genetics
Hydrogen-Ion Concentration
Kinetics
Models, Molecular
N-Acetylmuramoyl-L-alanine Amidase chemistry
N-Acetylmuramoyl-L-alanine Amidase genetics
Phylogeny
Protein Binding drug effects
Protein Conformation, alpha-Helical
Protein Conformation, beta-Strand
Protein Interaction Domains and Motifs
Recombinant Proteins chemistry
Recombinant Proteins genetics
Recombinant Proteins metabolism
Sequence Homology, Amino Acid
Sodium Chloride chemistry
Sodium Chloride pharmacology
Substrate Specificity
Viral Proteins chemistry
Viral Proteins genetics
Zinc chemistry
Zinc pharmacology
Bacteriophages chemistry
Endopeptidases metabolism
N-Acetylmuramoyl-L-alanine Amidase metabolism
Peptidoglycan metabolism
Vibrio parahaemolyticus virology
Viral Proteins metabolism
Subjects
Details
- Language :
- English
- ISSN :
- 1096-0279
- Volume :
- 188
- Database :
- MEDLINE
- Journal :
- Protein expression and purification
- Publication Type :
- Academic Journal
- Accession number :
- 34508857
- Full Text :
- https://doi.org/10.1016/j.pep.2021.105971