Proton-Translocating Adenosine Triphosphatase Activity in Lactic Acid Bacterial

Nannen, Nancy L.; Hutkins, Robert W.

doi:10.3168/jds.s0022-0302(91)78220-9

Cited by 67 publications

(49 citation statements)

References 12 publications

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“…bulgaricus, the onset of the constant ApH at pH,, 6 coincides with the optimal pH,, for growth of 5-8 (Beal et al, 1989), indicating a tighter control of pH, at acidic pH,,. This is further supported by the significantly lower pH optimum for H+-ATPase (5-0) of an acid-tolerant bacterium, Lactobacillus casei (Bender & Marquis, 1987), compared to the less acidtolerant Streptococcus therrnophilus and Lactococcus lactis with a pH optimum of 7-5 (Nannen & Hutkins, 1991b). The presence of short fatty acid anions such as lactate and acetate is also involved in the regulation of pHi in acid-tolerant bacteria (Russell, 1991a;Cook & Russell, 1994); however, this factor has not been investigated in the present work.…”

Section: Calibration Curve For Measuring Phi In Single Cellsmentioning

confidence: 82%

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Use of fluorescence ratio imaging for intracellular pH determination of individual bacterial cells in mixed cultures

Siegumfeldt¹,

Rechinger²,

Jakobsen³

1999

Microbiology

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The development of a rapid method for measuring intracellular pH (pH,) in single bacterial cells is described. Lactobacillus delbrueckii subsp. bulgaricus and Listeria innocua were used as test organisms. The method is based upon fluorescence microscopy and ratio imaging of cells stained with carboxyfluorescein succinimidyl ester. After staining, the bacteria were immobilized on a membrane filter and transferred to a closed perfusion chamber, allowing control of the cell environment during analysis. The set-up was optimized with regard to the use of neutral-density filters and background subtraction, for determining the excitation ratio 490 nm/435 nm (R49,,,435) independent of the excitation light intensity, and to reduce photobleaching. This allowed for time-lapse studies with multiple exposures.To study the pH, of Lb. delbrueckii subsp. bulgaricus and L. innocua in response to different extracellular pH (pH,,) values, an in vivo calibration curve was constructed in the pH, range 5-0-8-5. Distinct differences between the two cultures were observed. L. innocua maintained a near-neutral pH, almost independently of pH, (5-&8-0), whereas the pH, of Lb. delbmeckii subsp.bulgaricus decreased with decreasing pH, ,. In pure and mixed cultures a t pH, 5-0, the pH, values of Lb. delbmeckii subsp. bulgaricus and L. innocua were 6120.2 and 7.5202, respectively. This difference in pH, may explain how Lb. delbrueckii subsp. bulgaricus obtains a competitive advantage over L. innocua at low pH, , .Keywords : intracellular pH, ratio imaging, single cells, Lactobacillus delbrueckii subsp. bulgaricus, Listeria innocua INTRODUCTIONMost bacteria maintain an intracellular pH (pH,) close to neutral within fairly narrow limits (Padan et al., 19Sl), because this enables metabolic reactions to occur even under unfavoura ble extracellular pH (pH,,) conditions. Bacteria can be divided into three groups with regard to pH homeostasis : neutrophiles, acidophiles and alkalophiles. These groups differ in requirement for pH,,, but all groups maintain a pH, between 6.5 and 9-5 (White, 1995). Acidophiles maintain a large gradient between pH, and pH,, (ApH), but are restricted to growth in very acidic environments due to an inverted membrane potential (White, 1995 group of acid-tolerant fermentative bacteria grows at pH values ranging from neutral to pH 3-5 (Kashket, 1987;McDonald et al., 1990; Russell, 1991a). This group consists of certain ruminal bacteria (Russell, 1991b) and various species of lactic acid bacteria (Kashket, 1987). A common feature is the ability to decrease their pH, with pH,, during growth (Russell & Hino, 1985;Nannen & Hutkins, 1991a;Cook & Russell, 1994), and therefore this group of bacteria does not comply with the conventional classification of pH homeostasis. The pronounced organic acid production of these bacteria creates an environment unfavourable for most other organisms (Russell, 1992), which is the basis of many methods of food preservation by fermentation.Food fermentations are often carried out by a concerte...

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Section: Calibration Curve For Measuring Phi In Single Cellsmentioning

confidence: 82%

“…H+-ATPase has been shown to be the key enzyme in regulating pH, in Enterococcus faecalis (Kobayashi et al, 1986), and it is also assumed to regulate pH, in lactic acid bacteria (Nannen & Hutkins, 1991b). In L6.…”

Section: Calibration Curve For Measuring Phi In Single Cellsmentioning

confidence: 99%

Use of fluorescence ratio imaging for intracellular pH determination of individual bacterial cells in mixed cultures

Siegumfeldt¹,

Rechinger²,

Jakobsen³

1999

Microbiology

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“…A fraction of this energy is probably rerouted towards ATPase activity in order to increase the DpH under acidic conditions. This enzyme, found to be induced at low pH (Kobayashi et al, 1986;Nannen & Hutkins, 1991b;O'Sullivan & Condon, 1999), is able to extrude protons coupled to ATP consumption and thereby contributes to the maintenance of a favourable cellular pH.…”

Section: Discussion Acid Stress Damagesmentioning

confidence: 99%

Transcriptional, translational and metabolic regulation of glycolysis in Lactococcus lactis subsp. cremoris MG 1363 grown in continuous acidic cultures

2003

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The physiological behaviour of Lactococcus lactis subsp. cremoris MG 1363 was characterized in continuous culture under various acidic conditions (pH 4·7–6·6). Biomass yield was diminished in cultures with low pH and the energy dedicated to maintenance increased due to organic acid inhibition and cytoplasmic acidification. Under such acidic conditions, the specific rate of glucose consumption by the bacterium increased, thereby enhancing energy supply. This acceleration of glycolysis was regulated by both an increase in the concentrations of glycolytic enzymes (hierarchical regulation) and the specific modulation of enzyme activities (metabolic regulation). However, when the inhibitory effect of intracellular pH on enzyme activity was taken into account in the model of regulation, metabolite regulation was shown to be the dominant factor controlling pathway flux. The changes in glycolytic enzyme concentrations were not correlated directly to modifications in transcript concentrations. Analyses of the relative contribution of the phenomena controlling enzyme synthesis indicated that translational regulation had a major influence compared to transcriptional regulation. An increase in the translation efficiency was accompanied by an important decrease of total cellular RNA concentrations, confirming that the translation apparatus of L. lactis was optimized under acid stress conditions.

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“…For many bacteria it has been shown that treatment with acid and a subsequent decrease in the intracellular pH are accompanied by an increase in the amount of the F 1 F 0 -ATPase (2,19,21,30,31). This phenomenon has been studied most extensively in S. faecalis (2,17), in L. lactis (18,19), and in L. acidophilus (21), in which the cytoplasmic pH is maintained by the amount and activity of the H ϩ -ATPase, which catalyzes the ATP-driven translocation of protons from the cytoplasm.…”

Section: Discussionmentioning

confidence: 99%

Bifidobacterium lactisDSM 10140: Identification of theatp(atpBEFHAGDC) Operon and Analysis of Its Genetic Structure, Characteristics, and Phylogeny

Ventura

Canchaya

Sinderen³

et al. 2004

Appl Environ Microbiol

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The atp operon is highly conserved among eubacteria, and it has been considered a molecular marker as an alternative to the 16S rRNA gene. PCR primers were designed from the consensus sequences of the atpD gene to amplify partial atpD sequences from 12 Bifidobacterium species and nine Lactobacillus species. All PCR products were sequenced and aligned with other atpD sequences retrieved from public databases. Genes encoding the subunits of the F 1 F 0 -ATPase of Bifidobacterium lactis DSM 10140 (atpBEFHAGDC) were cloned and sequenced. The deduced amino acid sequences of these subunits showed significant homology with the sequences of other organisms. We identified specific sequence signatures for the genus Bifidobacterium and for the closely related taxa Bifidobacterium lactis and Bifidobacterium animalis and Lactobacillus gasseri and Lactobacillus johnsonii, which could provide an alternative to current methods for identification of lactic acid bacterial species. Northern blot analysis showed that there was a transcript at approximately 7.3 kb, which corresponded to the size of the atp operon, and a transcript at 4.5 kb, which corresponded to the atpC, atpD, atpG, and atpA genes. The transcription initiation sites of these two mRNAs were mapped by primer extension, and the results revealed no consensus promoter sequences. Phylogenetic analysis of the atpD genes demonstrated that the Lactobacillus atpD gene clustered with the genera Listeria, Lactococcus, Streptococcus, and Enterococcus and that the higher G؉C content and highly biased codon usage with respect to the genome average support the hypothesis that there was probably horizontal gene transfer. The acid inducibility of the atp operon of B. lactis DSM 10140 was verified by slot blot hybridization by using RNA isolated from acid-treated cultures of B. lactis DSM 10140. The rapid increase in the level of atp operon transcripts upon exposure to low pH suggested that the ATPase complex of B. lactis DSM 10140 was regulated at the level of transcription and not at the enzyme assembly step.

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Proton-Translocating Adenosine Triphosphatase Activity in Lactic Acid Bacterial

Cited by 67 publications

References 12 publications

Use of fluorescence ratio imaging for intracellular pH determination of individual bacterial cells in mixed cultures

Use of fluorescence ratio imaging for intracellular pH determination of individual bacterial cells in mixed cultures

Transcriptional, translational and metabolic regulation of glycolysis in Lactococcus lactis subsp. cremoris MG 1363 grown in continuous acidic cultures

Bifidobacterium lactisDSM 10140: Identification of theatp(atpBEFHAGDC) Operon and Analysis of Its Genetic Structure, Characteristics, and Phylogeny

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