Localization and functional role of the pseudomonas bacteriophage 2 depolymerase

Castillo, Francisco; Bartell, Pasquale F.

doi:10.1128/jvi.18.2.701-708.1976

Cited by 13 publications

(5 citation statements)

References 31 publications

(29 reference statements)

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“…Only the latter triggers irreversible conformational changes and ultimately leads to DNA ejection (Casjens and Molineux, ). In the case of phages featuring depolymerases, the EPS material itself likely represents the primary receptor (Castillo and Bartell, ; Hughes et al ., ). This hypothesis agrees well with our findings that adsorption of L1 is directly dependent on the presence of EPS on the cell surface and that phage binding did not occur with EPS‐deficient mutants or DpoL1 pretreated cells, or when DpoL1 was blocked by antibodies.…”

Section: Discussionmentioning

confidence: 99%

The tail‐associated depolymerase of Erwinia amylovora phage L1 mediates host cell adsorption and enzymatic capsule removal, which can enhance infection by other phage

Born

Fieseler

Klumpp

et al. 2013

Environmental Microbiology

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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.

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Section: Discussionmentioning

confidence: 99%

The tail‐associated depolymerase of Erwinia amylovora phage L1 mediates host cell adsorption and enzymatic capsule removal, which can enhance infection by other phage

Born

Fieseler

Klumpp

et al. 2013

Environmental Microbiology

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“…This phage displays depolymerase activity owing to the constituent enzyme. After treatment of bacterial cells with purified enzyme isolated from virion, bacteria P. aerurinosa lose phage sensitivity (Castillo and Bartell, 1976).…”

Section: Bacteriophage Receptors On Cell Surfacementioning

confidence: 99%

Bacteriophage Receptors, Mechanisms of Phage Adsorption and Penetration into Host Cell

et al. 2010

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Bacteriophages are an attractive tool for application in the therapy of bacterial infections, for biological control of bacterial contamination of foodstuffs in the alimentary industry, in plant protection, for control of water-borne pathogens, and control of environmental microflora. This review is mainly focused on structures governing phage recognition of host cell and mechanisms of phage adsorption and penetration into microbial cell.

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“…The contour plots generated in this study on the basis of data obtained were not perfectly elliptical, but horizontal which indicated less interaction occurring between the independent variables, ammonium chloride, and magnesium sulfate. The inorganic salts, ammonium chloride, and magnesium sulfate may be required as cofactors for efficient enzyme production as reported earlier for various phage depolymerases .…”

Section: Discussionmentioning

confidence: 99%

Aeromonas punctataderived depolymerase that disrupts the integrity ofKlebsiella pneumoniaecapsule: optimization of depolymerase production

Bansal

Soni

Harjai

et al. 2013

J. Basic Microbiol.

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Formation of dense, highly hydrated biofilm structures pose a risk for public and environmental health. Extracellular polymeric substances encompassing biofilms offer 1000-fold greater resistance as compared to the planktonic cells. Using enzymes as anti-biofouling agents, will improve penetration of antimicrobials and increase susceptibility of biofilms to components of immune system. The challenge of using enzymes derived from unrelated bacteria for the degradation of capsular matrix of Klebsiella pneumoniae has not been dealt in the past. Thus, statistical optimization was done to enhance depolymerase production by Aeromonas punctata, directed against the exopolysaccharide matrix of Klebsiella pneumoniae B5055, capable of substituting the available phage borne depolymerase enzyme. Optimization via central composite design (CCD) resulted in 16-fold enhancement in depolymerase yield (166.65 µmoles ml(-1) min(-1) ) over unoptimized medium. Out of the 19 variables, media composition giving maximum expression levels of the enzyme consisted of 1 mg ml(-1) galactose and ammonium chloride, 1.5 mg ml(-1) each of capsular polysaccharide (CPS) and magnesium sulfate. Tryptic peptide analysis of the purified 29 kDa band by Matrix assisted laser desorption ionization-time of flight (MALDI-TOF) showed a high homology with a protein of unknown function from Aeromonas cavaie Ae398. Further improvements in the enzyme can lead to its successful development as prophylactic and/or a therapeutic agent.

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Localization and functional role of the pseudomonas bacteriophage 2 depolymerase

Cited by 13 publications

References 31 publications

The tail‐associated depolymerase of Erwinia amylovora phage L1 mediates host cell adsorption and enzymatic capsule removal, which can enhance infection by other phage

The tail‐associated depolymerase of Erwinia amylovora phage L1 mediates host cell adsorption and enzymatic capsule removal, which can enhance infection by other phage

Bacteriophage Receptors, Mechanisms of Phage Adsorption and Penetration into Host Cell

Aeromonas punctataderived depolymerase that disrupts the integrity ofKlebsiella pneumoniaecapsule: optimization of depolymerase production

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