Cell wall hydrolase as a surface-associated protein target for the specific detection of Lactobacillus rhamnosus using flow cytometry

ABSTRACT A novel cell wall hydrolase encoded by the murA gene of Listeria monocytogenes is reported here. Mature MurA is a 66-kDa cell surface protein that is recognized by the well-characterized L. monocytogenes-specific monoclonal antibody EM-7G1. MurA displays two characteristic features: (i) an N-terminal domain with homology to muramidases from several gram-positive bacterial species and (ii) four copies of a cell wall-anchoring LysM repeat motif present within its C-terminal domain. Purified recombinant MurA produced in Escherichia coli was confirmed to be an authentic cell wall hydrolase with lytic properties toward cell wall preparations of Micrococcus lysodeikticus. An isogenic mutant with a deletion of murA that lacked the 66-kDa cell wall hydrolase grew as long chains during exponential growth. Complementation of the mutant strain by chromosomal reintegration of the wild-type gene restored expression of this murein hydrolase activity and cell separation levels to those of the wild-type strain. Studies reported herein suggest that the MurA protein is involved in generalized autolysis of L. monocytogenes.

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Purification and characterization of a cell wall hydrolase encoded by the cw1A gene ofBacillus subtilis

FEMS Microbiology Letters ◽

10.1111/j.1574-6968.1991.tb04703.x ◽

1991 ◽

Vol 81 (1) ◽

pp. 9-13 ◽

Cited By ~ 11

Author(s):

Akio Kuroda ◽

Masafumi Imazeki ◽

Junichi Sekiguchi

Keyword(s):

Cell Wall ◽

Purification And Characterization ◽

Ofbacillus Subtilis ◽

Cell Wall Hydrolase ◽

A Cell

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Molecular cloning of a sporulation-specific cell wall hydrolase gene of Bacillus subtilis.

Journal of Bacteriology ◽

10.1128/jb.175.19.6260-6268.1993 ◽

1993 ◽

Vol 175 (19) ◽

pp. 6260-6268 ◽

Cited By ~ 46

Author(s):

A Kuroda ◽

Y Asami ◽

J Sekiguchi

Keyword(s):

Cell Wall ◽

Bacillus Subtilis ◽

Molecular Cloning ◽

Specific Cell ◽

Cell Wall Hydrolase

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Antimicrobial Activity of Exebacase (Lysin CF-301) against the Most Common Causes of Infective Endocarditis

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.01078-19 ◽

2019 ◽

Vol 63 (10) ◽

Cited By ~ 7

Author(s):

Aubrey Watson ◽

Jun Taek Oh ◽

Karen Sauve ◽

Patricia A. Bradford ◽

Cara Cassino ◽

...

Keyword(s):

Antimicrobial Activity ◽

Cell Wall ◽

Infective Endocarditis ◽

Broth Microdilution ◽

Coagulase Negative Staphylococci ◽

Content Type ◽

Microdilution Method ◽

Cell Wall Hydrolase ◽

Broth Microdilution Method ◽

Common Causes

ABSTRACT Exebacase, a recombinantly produced lysin (cell wall hydrolase), and comparator antibiotics were tested by the broth microdilution method against strain sets of Staphylococcus and Streptococcus spp., which are the most common causes of infective endocarditis in humans. Exebacase was active against all Staphylococcus spp. tested, including S. aureus and coagulase-negative staphylococci (MIC50/90, 0.5/1 μg/ml). Activity against Streptococcus spp. was variable, with S. pyogenes, S. agalactiae, and S. dysgalactiae (MIC50/90, 1/2 μg/ml) among the most susceptible.

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