Functional Roles of Aggregation-Promoting-Like Factor in Stress Tolerance and Adherence of Lactobacillus acidophilus NCFM
Open Access
- 1 August 2010
- journal article
- Published by American Society for Microbiology in Applied and Environmental Microbiology
- Vol. 76 (15), 5005-5012
- https://doi.org/10.1128/aem.00030-10
Abstract
Aggregation-promoting factors (Apf) are secreted proteins that have been associated with a diverse number of functional roles in lactobacilli, including self-aggregation, the bridging of conjugal pairs, coaggregation with other commensal or pathogenic bacteria, and maintenance of cell shape. In silico genome analysis of Lactobacillus acidophilus NCFM identified LBA0493 as a 696-bp apf gene that encodes a putative 21-kDa Apf protein. Transcriptional studies of NCFM during growth in milk showed apf to be one of the most highly upregulated genes in the genome. In the present study, reverse transcriptase-quantitative PCR (RT-QPCR) analysis revealed that the apf gene was highly induced during the stationary phase compared to that during the logarithmic phase. To investigate the functional role of Apf in NCFM, an Δ apf deletion mutant was constructed. The resulting Δ apf mutant, NCK2033, did not show a significant difference in cell morphology or growth compared to that of the NCFMΔ upp reference strain, NCK1909. The autoaggregation phenotype of NCK2033 in planktonic culture was unaffected. Additional phenotypic assays revealed that NCK2033 was more susceptible to treatments with oxgall bile and sodium dodecyl sulfate (SDS). Survival rates of NCK2033 decreased when stationary-phase cells were exposed to simulated small-intestinal and gastric juices. Furthermore, NCK2033 in the stationary phase showed a reduction of in vitro adherence to Caco-2 intestinal epithelial cells, mucin glycoproteins, and fibronectin. The data suggest that the Apf-like proteins may contribute to the survival of L. acidophilus during transit through the digestive tract and, potentially, participate in the interactions with the host intestinal mucosa.Keywords
This publication has 47 references indexed in Scilit:
- Role of Transporter Proteins in Bile Tolerance ofLactobacillus acidophilusApplied and Environmental Microbiology, 2009
- Development and Application of a upp -Based Counterselective Gene Replacement System for the Study of the S-Layer Protein SlpX of Lactobacillus acidophilus NCFMApplied and Environmental Microbiology, 2009
- Clustal W and Clustal X version 2.0Bioinformatics, 2007
- Aggregating Phenotype in Lactobacillus crispatus Determines Intestinal Colonization and TLR2 and TLR4 Modulation in Murine Colonic MucosaClinical and Vaccine Immunology, 2007
- Characterization of a Novel Bile-Inducible Operon Encoding a Two-Component Regulatory System inLactobacillus acidophilusJournal of Bacteriology, 2007
- Identification of a Putative Operon Involved in Fructooligosaccharide Utilization by Lactobacillus paracaseiApplied and Environmental Microbiology, 2006
- Improved Prediction of Signal Peptides: SignalP 3.0Journal of Molecular Biology, 2004
- Essential Role for the gtfA Gene Encoding a Putative Glycosyltransferase in the Adherence of Porphyromonas gingivalisInfection and Immunity, 2004
- Characterization of the aggregation promoting factor from Lactobacillus gasseri , avaginal isolateJournal of Applied Microbiology, 1997
- Engineering hybrid genes without the use of restriction enzymes: gene splicing by overlap extensionGene, 1989