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

DOI: 10.1002/pro.4585

Authors: Andrius Jasilionis (Lund University) , Magdalena Plotka (University of Gdansk) , Lei Wang (University of Stuttgart) , Sebastian Dorawa (University of Gdansk) , Joanna Lange (Bio-Prodict BV) , Hildegard Watzlawick (University of Stuttgart) , Tom Van Den Bergh (Bio-Prodict BV) , Bas Vroling (Bio-Prodict BV) , Josef Altenbuchner (University of Stuttgart) , Anna-Karina Kaczorowska (University of Gdansk) , Ehmke Pohl (Durham University) , Tadeusz Kaczorowski (University of Gdansk) , Eva Nordberg Karlsson (Lund University) , Stefanie Freitag-Pohl (Durham University)
Co-authored by industrial partner: No

Type: Journal Paper
Journal: Protein Science

State: Published (Approved)
Published: January 2023
Diamond Proposal Number(s): 18598

Abstract: 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 to protein structures of similar proteins. The functional characterisation of AmiP demonstrates high efficiency of catalytic activity and broad substrate specificity towards 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 characterised 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.

Journal Keywords: peptidoglycan lytic amidases; Amidase_3 catalytic domain; Thermus prophage; adaptation; thermoactivity; thermostability

Diamond Keywords: Bacteriophages; Viruses

Subject Areas: Biology and Bio-materials, Chemistry, Medicine


Instruments: I03-Macromolecular Crystallography

Added On: 02/02/2023 09:50

Discipline Tags:

Antibiotic Resistance Biotechnology Health & Wellbeing Biochemistry Chemistry Structural biology Engineering & Technology Drug Discovery Life Sciences & Biotech

Technical Tags:

Diffraction Macromolecular Crystallography (MX)