Functional analysis of D-alanylation of lipoteichoic acid in the probiotic strain Lactobacillus rhamnosus GG

Appl Environ Microbiol. 2007 Jun;73(11):3595-604. doi: 10.1128/AEM.02083-06. Epub 2007 Apr 13.

Abstract

Lipoteichoic acid (LTA) is a macroamphiphile molecule which performs several functions in gram-positive bacteria, such as maintenance of cell wall homeostasis. D-alanylation of LTA requires the proteins encoded by the dlt operon, and this process is directly related to the charge properties of this polymer strongly contributing to its function. The insertional inactivation of dltD of the probiotic strain Lactobacillus rhamnosus GG (ATCC 53103) resulted in the complete absence of D-alanyl esters in the LTA as confirmed by nuclear magnetic resonance analysis. This was reflected in modifications of the bacterial cell surface properties. The dltD strain showed 2.4-fold-increased cell length, a low survival capacity in response to gastric juice challenge, an increased sensitivity to human beta-defensin-2, an increased rate of autolysis, an increased capacity to initiate growth in the presence of an anionic detergent, and a decreased capacity to initiate growth in the presence of cationic peptides compared to wild-type results. However, in vitro experiments revealed no major differences for adhesion to human intestinal epithelial cells, biofilm formation, and immunomodulation. These properties are considered to be important for probiotics. The role of the dlt operon in lactobacilli is discussed in view of these results.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alanine / metabolism*
  • Anti-Bacterial Agents / pharmacology
  • Bacterial Adhesion / genetics
  • Bacterial Proteins / genetics
  • Bacteriolysis / genetics
  • Biofilms / growth & development
  • Cell Line
  • Cell Wall / chemistry
  • Cell Wall / metabolism
  • Cytokines / immunology
  • DNA, Bacterial / chemistry
  • DNA, Bacterial / genetics
  • Epithelial Cells / immunology
  • Epithelial Cells / microbiology
  • Gastric Juice / microbiology
  • Gene Deletion
  • Humans
  • Lacticaseibacillus rhamnosus / drug effects
  • Lacticaseibacillus rhamnosus / genetics
  • Lacticaseibacillus rhamnosus / immunology
  • Lacticaseibacillus rhamnosus / metabolism*
  • Lipopolysaccharides / chemistry
  • Lipopolysaccharides / metabolism*
  • Magnetic Resonance Spectroscopy
  • Microbial Viability / genetics
  • Microscopy, Electron, Transmission
  • Molecular Sequence Data
  • Mutagenesis, Insertional
  • Nisin / pharmacology
  • Polymyxin B / pharmacology
  • Probiotics / chemistry
  • Probiotics / metabolism*
  • Sequence Analysis, DNA
  • Teichoic Acids / chemistry
  • Teichoic Acids / metabolism*
  • Thiolester Hydrolases / genetics
  • beta-Defensins / immunology

Substances

  • Anti-Bacterial Agents
  • Bacterial Proteins
  • Cytokines
  • DEFB4A protein, human
  • DNA, Bacterial
  • Lipopolysaccharides
  • Teichoic Acids
  • beta-Defensins
  • Nisin
  • lipoteichoic acid
  • DltD protein, bacteria
  • Thiolester Hydrolases
  • Polymyxin B
  • Alanine

Associated data

  • GENBANK/DQ906101