Knockout of the alanine racemase gene in Lactobacillus plantarum results in septation defects and cell wall perforation

E. Palumbo, C.F. Favier, M. Deghorain, P.S. Cocconcelli, C. Grangette, A.M.E. Mercenier, E.E. Vaughan, P. Hols

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Abstract

A stable mutant of Lactobacillus plantarum deficient in alanine racemase (Alr) was constructed by two successive homologous recombination steps. When the mutant was supplemented with D-alanine, growth and viability were unaffected. Surprisingly, deprivation Of D-alanine during exponential growth did not result in a rapid and extensive lysis as observed in Alr-deficient strains of Escherichia coli or Bacillus subtilis. Rather, the starved mutant cells underwent a growth arrest and were gradually affected in viability with a decrease in colony forming units over 99% in less than 24 h. Additionally, fluorescent techniques demonstrated a loss of cell envelope integrity in the starved cells. Prolonged D-alanine starvation resulted in cells with an aberrant morphology. Scanning and transmission electron microscopy analyses revealed an increase in cell length, deficiencies in septum formation, thinning of the cell envelope and perforation of the cell wall in the septum region. We discuss the involvement of peptidoglycan hydrolases in these phenotypic defects in the context of the crucial role played by D-alanine in peptidoglycan biosynthesis and teichoic acids substitution. (C) 2004 Federation of European Microbiological Societies. Published by Elsevier B.V. All rights reserved.
Original languageEnglish
Pages (from-to)131-138
JournalFEMS Microbiology Letters
Volume233
Issue number1
DOIs
Publication statusPublished - 2004

Fingerprint

Alanine Racemase
Lactobacillus plantarum
Cell Wall
Alanine
Genes
Growth
N-Acetylmuramoyl-L-alanine Amidase
Teichoic Acids
Scanning Transmission Electron Microscopy
Peptidoglycan
Homologous Recombination
Starvation
Bacillus subtilis
Stem Cells
Escherichia coli

Keywords

  • lactic-acid bacteria
  • lipoteichoic acid
  • bacillus-subtilis
  • escherichia-coli
  • teichoic-acid
  • lysis
  • methicillin
  • autolysis
  • mutants
  • impact

Cite this

Palumbo, E., Favier, C. F., Deghorain, M., Cocconcelli, P. S., Grangette, C., Mercenier, A. M. E., ... Hols, P. (2004). Knockout of the alanine racemase gene in Lactobacillus plantarum results in septation defects and cell wall perforation. FEMS Microbiology Letters, 233(1), 131-138. https://doi.org/10.1016/j.femsle.2004.02.001
Palumbo, E. ; Favier, C.F. ; Deghorain, M. ; Cocconcelli, P.S. ; Grangette, C. ; Mercenier, A.M.E. ; Vaughan, E.E. ; Hols, P. / Knockout of the alanine racemase gene in Lactobacillus plantarum results in septation defects and cell wall perforation. In: FEMS Microbiology Letters. 2004 ; Vol. 233, No. 1. pp. 131-138.
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abstract = "A stable mutant of Lactobacillus plantarum deficient in alanine racemase (Alr) was constructed by two successive homologous recombination steps. When the mutant was supplemented with D-alanine, growth and viability were unaffected. Surprisingly, deprivation Of D-alanine during exponential growth did not result in a rapid and extensive lysis as observed in Alr-deficient strains of Escherichia coli or Bacillus subtilis. Rather, the starved mutant cells underwent a growth arrest and were gradually affected in viability with a decrease in colony forming units over 99{\%} in less than 24 h. Additionally, fluorescent techniques demonstrated a loss of cell envelope integrity in the starved cells. Prolonged D-alanine starvation resulted in cells with an aberrant morphology. Scanning and transmission electron microscopy analyses revealed an increase in cell length, deficiencies in septum formation, thinning of the cell envelope and perforation of the cell wall in the septum region. We discuss the involvement of peptidoglycan hydrolases in these phenotypic defects in the context of the crucial role played by D-alanine in peptidoglycan biosynthesis and teichoic acids substitution. (C) 2004 Federation of European Microbiological Societies. Published by Elsevier B.V. All rights reserved.",
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author = "E. Palumbo and C.F. Favier and M. Deghorain and P.S. Cocconcelli and C. Grangette and A.M.E. Mercenier and E.E. Vaughan and P. Hols",
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Palumbo, E, Favier, CF, Deghorain, M, Cocconcelli, PS, Grangette, C, Mercenier, AME, Vaughan, EE & Hols, P 2004, 'Knockout of the alanine racemase gene in Lactobacillus plantarum results in septation defects and cell wall perforation', FEMS Microbiology Letters, vol. 233, no. 1, pp. 131-138. https://doi.org/10.1016/j.femsle.2004.02.001

Knockout of the alanine racemase gene in Lactobacillus plantarum results in septation defects and cell wall perforation. / Palumbo, E.; Favier, C.F.; Deghorain, M.; Cocconcelli, P.S.; Grangette, C.; Mercenier, A.M.E.; Vaughan, E.E.; Hols, P.

In: FEMS Microbiology Letters, Vol. 233, No. 1, 2004, p. 131-138.

Research output: Contribution to journalArticleAcademicpeer-review

TY - JOUR

T1 - Knockout of the alanine racemase gene in Lactobacillus plantarum results in septation defects and cell wall perforation

AU - Palumbo, E.

AU - Favier, C.F.

AU - Deghorain, M.

AU - Cocconcelli, P.S.

AU - Grangette, C.

AU - Mercenier, A.M.E.

AU - Vaughan, E.E.

AU - Hols, P.

PY - 2004

Y1 - 2004

N2 - A stable mutant of Lactobacillus plantarum deficient in alanine racemase (Alr) was constructed by two successive homologous recombination steps. When the mutant was supplemented with D-alanine, growth and viability were unaffected. Surprisingly, deprivation Of D-alanine during exponential growth did not result in a rapid and extensive lysis as observed in Alr-deficient strains of Escherichia coli or Bacillus subtilis. Rather, the starved mutant cells underwent a growth arrest and were gradually affected in viability with a decrease in colony forming units over 99% in less than 24 h. Additionally, fluorescent techniques demonstrated a loss of cell envelope integrity in the starved cells. Prolonged D-alanine starvation resulted in cells with an aberrant morphology. Scanning and transmission electron microscopy analyses revealed an increase in cell length, deficiencies in septum formation, thinning of the cell envelope and perforation of the cell wall in the septum region. We discuss the involvement of peptidoglycan hydrolases in these phenotypic defects in the context of the crucial role played by D-alanine in peptidoglycan biosynthesis and teichoic acids substitution. (C) 2004 Federation of European Microbiological Societies. Published by Elsevier B.V. All rights reserved.

AB - A stable mutant of Lactobacillus plantarum deficient in alanine racemase (Alr) was constructed by two successive homologous recombination steps. When the mutant was supplemented with D-alanine, growth and viability were unaffected. Surprisingly, deprivation Of D-alanine during exponential growth did not result in a rapid and extensive lysis as observed in Alr-deficient strains of Escherichia coli or Bacillus subtilis. Rather, the starved mutant cells underwent a growth arrest and were gradually affected in viability with a decrease in colony forming units over 99% in less than 24 h. Additionally, fluorescent techniques demonstrated a loss of cell envelope integrity in the starved cells. Prolonged D-alanine starvation resulted in cells with an aberrant morphology. Scanning and transmission electron microscopy analyses revealed an increase in cell length, deficiencies in septum formation, thinning of the cell envelope and perforation of the cell wall in the septum region. We discuss the involvement of peptidoglycan hydrolases in these phenotypic defects in the context of the crucial role played by D-alanine in peptidoglycan biosynthesis and teichoic acids substitution. (C) 2004 Federation of European Microbiological Societies. Published by Elsevier B.V. All rights reserved.

KW - lactic-acid bacteria

KW - lipoteichoic acid

KW - bacillus-subtilis

KW - escherichia-coli

KW - teichoic-acid

KW - lysis

KW - methicillin

KW - autolysis

KW - mutants

KW - impact

U2 - 10.1016/j.femsle.2004.02.001

DO - 10.1016/j.femsle.2004.02.001

M3 - Article

VL - 233

SP - 131

EP - 138

JO - FEMS Microbiology Letters

JF - FEMS Microbiology Letters

SN - 0378-1097

IS - 1

ER -