AmiP from hyperthermophilic Thermus parvatiensis prophage is a thermoactive and ultrathermostable peptidoglycan lytic amidase

Bacteriophages encode a wide variety of cell wall disrupting enzymes that aid the viral escape in the final stages of infection. These lytic enzymes have accumulated notable interest due to their potential as novel antibacterials for infection treatment caused by multiple‐drug resistant bacteria. He...

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Published in:	Protein science 2023-03, Vol.32 (3), p.e4585-n/a
Main Authors:	Jasilionis, Andrius, Plotka, Magdalena, Wang, Lei, Dorawa, Sebastian, Lange, Joanna, Watzlawick, Hildegard, van den Bergh, Tom, Vroling, Bas, Altenbuchner, Josef, Kaczorowska, Anna‐Karina, Pohl, Ehmke, Kaczorowski, Tadeusz, Nordberg Karlsson, Eva, Freitag‐Pohl, Stefanie
Format:	Article
Language:	eng
Subjects:	adaptation Amidase Amidase_3 catalytic domain Amidohydrolases - metabolism Bacteria Biochemistry and Molecular Biology Biokemi och molekylärbiologi Biologi Biological Sciences Biotechnology Catalytic activity Catalytic Domain Cell Wall Cell walls Crystal structure Drug resistance Enzymes Full‐length Paper Full‐length Papers Heat resistance High temperature Infections Lytic enzymes Melt temperature Mesophilic bacteria Models, Molecular Multidrug resistance N-Acetylmuramoyl-L-alanine Amidase - chemistry N-Acetylmuramoyl-L-alanine Amidase - metabolism Natural Sciences Naturvetenskap Peptidoglycan - metabolism peptidoglycan lytic amidases Peptidoglycans pH effects Phages Prophages - metabolism Protein folding Protein structure Proteins Secondary structure Sodium Chloride Structural analysis Structure-function relationships Substrate specificity Substrates thermoactivity Thermophilic bacteria thermostability Thermus prophage
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Summary:	Bacteriophages encode a wide variety of cell wall disrupting enzymes that aid the viral escape in the final stages of infection. These lytic enzymes have accumulated notable interest due to their potential as novel antibacterials for infection treatment caused by multiple‐drug resistant bacteria. Here, the detailed functional and structural characterization of Thermus parvatiensis prophage peptidoglycan lytic amidase AmiP, a globular Amidase_3 type lytic enzyme adapted to high temperatures is presented. The sequence and structure comparison with homologous lytic amidases reveals the key adaptation traits that ensure the activity and stability of AmiP at high temperatures. The crystal structure determined at a resolution of 1.8 Å displays a compact α/β‐fold with multiple secondary structure elements omitted or shortened compared with protein structures of similar proteins. The functional characterization of AmiP demonstrates high efficiency of catalytic activity and broad substrate specificity toward thermophilic and mesophilic bacteria strains containing Orn‐type or DAP‐type peptidoglycan. The here presented AmiP constitutes the most thermoactive and ultrathermostable Amidase_3 type lytic enzyme biochemically characterized with a temperature optimum at 85°C. The extraordinary high melting temperature Tm 102.6°C confirms fold stability up to approximately 100°C. Furthermore, AmiP is shown to be more active over the alkaline pH range with pH optimum at pH 8.5 and tolerates NaCl up to 300 mM with the activity optimum at 25 mM NaCl. This set of beneficial characteristics suggests that AmiP can be further exploited in biotechnology. PDB Code(s): 7B3N.
ISSN:	0961-8368 1469-896X 1469-896X