Title: The tail-associated depolymerase of Erwinia amylovoraphage L1 mediates host cell adsorption and enzymatic capsule removal, which can enhance infection by other phage
Authors : Born, Yannick
Fieseler, Lars
Klumpp, Jochen
Eugster, Marcel R.
Zurfluh, Katrin
Duffy, Brion
Loessner, Martin J.
Published in : Environmental Microbiology
Volume(Issue) : 16
Issue : 7
Pages : 2168
Pages to: 2180
Publisher / Ed. Institution : Wiley
Issue Date: 2014
License (according to publishing contract) : Licence according to publishing contract
Type of review: Peer review (Publication)
Language : English
Subjects : Bacterial adhesion; Erwinia amylovora; Escherichia coli; Host Specificity; Hydrogen-Ion concentration; Hydrolysis; Kinetics; Podoviridae; Bacterial polysaccharides; Tertiary protein structure; Recombinant proteins; Rosaceae; Viral proteins; Virion; Biological control agents
Subject (DDC) : 570: Biology
Abstract: The depolymerase enzyme (DpoL1) encoded by the T7-like phage L1 efficiently degrades amylovoran, an important virulence factor and major component of the extracellular polysaccharide (EPS) of its host, the plant pathogen Erwinia amylovora. Mass spectrometry analysis of hydrolysed EPS revealed that DpoL1 cleaves the galactose-containing backbone of amylovoran. The enzyme is most active at pH 6 and 50°C, and features a modular architecture. Removal of 180 N-terminal amino acids was shown not to affect enzyme activity. The C-terminus harbours the hydrolase activity, while the N-terminal domain links the enzyme to the phage particle. Electron microscopy demonstrated that DpoL1-specific antibodies cross-link phage particles at their tails, either lateral or frontal, and immunogold staining confirmed that DpoL1 is located at the tail spikes. Exposure of high-level EPS-producing Er. amylovora strain CFBP1430 to recombinant DpoL1 dramatically increased sensitivity to the Dpo-negative phage Y2, which was not the case for EPS-negative mutants or low-level EPS-producing Er. amylovora. Our findings indicate that enhanced phage susceptibility is based on enzymatic removal of the EPS capsule, normally a physical barrier to Y2 infection, and that use of DpoL1 together with the broad host range, virulent phage Y2 represents an attractive combination for biocontrol of fire blight.
Departement: Life Sciences and Facility Management
Organisational Unit: Institute of Food and Beverage Innovation (ILGI)
Publication type: Article in scientific Journal
DOI : 10.1111/1462-2920.12212
ISSN: 14622912
URI: https://digitalcollection.zhaw.ch/handle/11475/12267
Appears in Collections:Publikationen Life Sciences und Facility Management

Files in This Item:
There are no files associated with this item.

Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.