Effect of Tween 80 on the Morphology and Physiology of Lactobacillus salivarius Strain IV CL-37 Grown in a Chemostat under Glucose Limitation

Jacques, Nicholas A.; Hardy, Lyn; Knox, K. W.; Wicken, A. J.

doi:10.1099/00221287-119-1-195

Cited by 21 publications

(20 citation statements)

References 8 publications

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“…Triplicate continuous cultures of Streptococcus mutans LT11 (Tao et al, 1993) were grown under anaerobic conditions at a dilution rate (D) of 0?100±0?001 h 21 , at either pH 7?0±0?1 or pH 5?0±0?1, with glucose limitation as previously described (Jacques et al, 1979). DMM medium, devoid of mucin, was used, but modified to include adenine, guanine and uracil at 20 mg ml 21 , and both KH 2 PO 4 and K 2 HPO 4 at 15 mM (Sissons et al, 1991).…”

Section: Methodsmentioning

confidence: 99%

Stress-responsive proteins are upregulated in Streptococcus mutans during acid tolerance

2004

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Streptococcus mutans is an important pathogen in the initiation of dental caries as the bacterium remains metabolically active when the environment becomes acidic. The mechanisms underlying this ability to survive and proliferate at low pH remain an area of intense investigation. Differential two-dimensional electrophoretic proteome analysis of S. mutans grown at steady state in continuous culture at pH 7?0 or pH 5?0 enabled the resolution of 199 cellular and extracellular protein spots with altered levels of expression. Matrix-assisted laser desorption ionization time-of-flight mass spectrometry identified 167 of these protein spots. Sixty-one were associated with stress-responsive pathways involved in DNA replication, transcription, translation, protein folding and proteolysis. The 61 protein spots represented isoforms or cleavage products of 30 different proteins, of which 25 were either upregulated or uniquely expressed during acid-tolerant growth at pH 5?0. Among the unique and upregulated proteins were five that have not been previously identified as being associated with acid tolerance in S. mutans and/or which have not been studied in any detail in oral streptococci. These were the single-stranded DNA-binding protein, Ssb, the transcription elongation factor, GreA, the RNA exonuclease, polyribonucleotide nucleotidyltransferase (PnpA), and two proteinases, the ATP-binding subunit, ClpL, of the Clp family of proteinases and a proteinase encoded by the pep gene family with properties similar to the dipeptidase, PepD, of Lactobacillus helveticus. The identification of these and other differentially expressed proteins associated with an acid-tolerant-growth phenotype provides new information on targets for mutagenic studies that will allow the future assessment of their physiological significance in the survival and proliferation of S. mutans in low pH environments. INTRODUCTIONStreptococcus mutans is now well recognized as being associated with the initiation of dental caries, since its acid fermentation by-products can result in the demineralization of tooth enamel (Hamada & Slade, 1980;Harper & Loesche, 1984;Loesche, 1986;van Houte, 1994;van Ruyven et al., 2000). A key to the survival of S. mutans at low pH is its ability to maintain a transmembrane pH gradient (DpH), with the interior of the cell more alkaline. This is achieved by upregulation of a proton-translocating F 1 F 0 -ATPase that extrudes H + as the external environment becomes more acidic. This results in an increased use of ATP for H + extrusion and a consequent reduction in cell yield (Belli & Marquis, 1991;Hamilton & Buckley, 1991;Dashper & Reynolds, 1992;Quivey et al., 2001). A series of recent physiological, mutagenic and proteome studies (Quivey et al., 1995;Gutierrez et al., 1996 Gutierrez et al., , 1999Jayaraman et al., 1997; Hamilton & Svensäter, 1998;Hahn et al., 1999;Hanna et al., 2001; Kremer et al., 2001;Lemos et al., 2001;Li et al., 2002;Wilkins et al., 2002;Len et al., 2004), however, indicates that S. mutans regulates its pheno...

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Section: Methodsmentioning

confidence: 99%

Stress-responsive proteins are upregulated in Streptococcus mutans during acid tolerance

2004

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“…Cells from triplicate continuous cultures of S. mutans LT11 (Tao et al, 1993) grown at 37 uC to a steady state at pH 7?0±0?1 and at a dilution rate (D) of 0?100±0?001 h 21 under anaerobic conditions (95 % N 2 / 5 % CO 2 ) (Jacques et al, 1979) were used as an inoculum for biofilm-grown cells as well as for supplying control planktonic cultures for subsequent comparative proteome analyses. The use of a chemostat greatly reduced the biological variation that is inherent when growing bacteria in batch culture (Len et al, 2004a, b).…”

Section: Methodsmentioning

confidence: 99%

Up-regulation of competence- but not stress-responsive proteins accompanies an altered metabolic phenotype in Streptococcus mutans biofilms

et al. 2005

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Mature biofilm and planktonic cells of Streptococcus mutans cultured in a neutral pH environment were subjected to comparative proteome analysis. Of the 242 protein spots identified, 48 were significantly altered in their level of expression (P<0?050) or were unique to planktonic or biofilm-grown cells. Among these were four hypothetical proteins as well as proteins known to be associated with the maintenance of competence or found to possess a cin-box-like element upstream of their coding gene. Most notable among the non-responsive genes were those encoding the molecular chaperones DnaK, GroEL and GroES, which are considered to be up-regulated by sessile growth. Analysis of the rest of the proteome indicated that a number of cellular functions associated with carbon uptake and cell division were down-regulated. The data obtained were consistent with the hypothesis that a reduction in the general growth rate of mature biofilms of S. mutans in a neutral pH environment is associated with the maintenance of transformation without the concomitant stress response observed during the transient state of competence in bacterial batch cultures.

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“…Because Lactobacillus rhamnosus hydrolyzes extracellular polysaccharide by glycohydrolase (Pham et al, 2000), enzymatic removal of those substances might be possible. The production of extracellular teichoic acids by Lactobacillus salivarius and Staphylococcus aureus is controlled by the addition of Tween 80 in the media used, or by the use of aero-anaerobic conditions (Jacques et al, 1980;Sadovskaya et al, 2005). Thus, to improve the survival rates during storage of these species when freeze-dried, future studies should examine which culture conditions suppress the production of polysaccharides and teichoic acids.…”

Section: Improving the Survival Of Freeze-dried Bacteria During Storagementioning

confidence: 99%

Survival of freeze-dried bacteria

Miyamoto‐Shinohara

Sukenobe

Imaizumi

et al. 2008

J. Gen. Appl. Microbiol.

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Effect of Tween 80 on the Morphology and Physiology of Lactobacillus salivarius Strain IV CL-37 Grown in a Chemostat under Glucose Limitation

Cited by 21 publications

References 8 publications

Stress-responsive proteins are upregulated in Streptococcus mutans during acid tolerance

Stress-responsive proteins are upregulated in Streptococcus mutans during acid tolerance

Up-regulation of competence- but not stress-responsive proteins accompanies an altered metabolic phenotype in Streptococcus mutans biofilms

Survival of freeze-dried bacteria

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