Enzymatic Treatment to Eliminate the Extracellular ATP for Improving the Detectability of Bacterial Intracellular ATP

Sakakibara, T.; Murakami, Seiji; Hattori, Noriaki; Nakajima, Mio; Imai, Kazuhiro

doi:10.1006/abio.1997.2217

Cited by 41 publications

(19 citation statements)

References 14 publications

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“…The ATP content of each sample tested was determined using an ATP assay reagent kit (Kikkoman Co. Ltd., Noda, Japan) based on the firefly luciferin-luciferase reaction (28). A solution of the indicator cells was prepared as follows.…”

Section: Methodsmentioning

confidence: 99%

Structural and Functional Differences in Two Cyclic Bacteriocins with the Same Sequences Produced by Lactobacilli

Kawai

Ishii

Arakawa

et al. 2004

Appl Environ Microbiol

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Lactobacillus gasseri LA39 and L. reuteri LA6 isolated from feces of the same human infant were found to produce similar cyclic bacteriocins (named gassericin A and reutericin 6, respectively) that cannot be distinguished by molecular weights or primary amino acid sequences. However, reutericin 6 has a narrower spectrum than gassericin A. In this study, gassericin A inhibited the growth of L. reuteri LA6, but reutericin 6 did not inhibit the growth of L. gasseri LA39. Both bacteriocins caused potassium ion efflux from indicator cells and liposomes, but the amounts of efflux and patterns of action were different. Although circular dichroism spectra of purified bacteriocins revealed that both antibacterial peptides are composed mainly of ␣-helices, the spectra of the bacteriocins did not coincide. The results of D-and L-amino acid composition analysis showed that two residues and one residue of D-Ala were detected among 18 Ala residues of gassericin A and reutericin 6, respectively. These findings suggest that the different D-alanine contents of the bacteriocins may cause the differences in modes of action, amounts of potassium ion efflux, and secondary structures. This is the first report that characteristics of native bacteriocins produced by wild lactobacillus strains having the same structural genes are influenced by a difference in D-amino acid contents in the molecules.

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Section: Methodsmentioning

confidence: 99%

Structural and Functional Differences in Two Cyclic Bacteriocins with the Same Sequences Produced by Lactobacilli

Kawai

Ishii

Arakawa

et al. 2004

Appl Environ Microbiol

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“…The ATP content of each sample tested was measured using an ATP-luciferase reaction kit (Lucifer HS; Kikkoman, Chiba, Japan), based on the firefly luciferin-luciferase reaction (19), and a luminescence-measuring instrument, TD-20/20 (Turner Designs, Sunnyvale, CA), according to the manufacturer's instructions. ATP content was measured in relative light units (RLU).…”

Section: Methodsmentioning

confidence: 99%

Quantitative Detection of Viable Bifidobacterium bifidum BF-1 Cells in Human Feces by Using Propidium Monoazide and Strain-Specific Primers

Fujimoto

Watanabe

2013

Appl Environ Microbiol

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We developed a PCR-based method to detect and quantify viable Bifidobacterium bifidum BF-1 cells in human feces. This method (PMA-qPCR) uses propidium monoazide (PMA) to distinguish viable from dead cells and quantitative PCR using a BF-1-specific primer set designed from the results of randomly amplified polymorphic DNA analysis. During long-term culture (10 days), the number of viable BF-1 cells detected by counting the number of CFU on modified MRS agar, by measuring the ATP contents converted to CFU, and by using PMA-qPCR decreased from about 10 10 to 10 6 cells/ml; in contrast, the total number of (viable and dead) BF-1 cells detected by counting 4=,6-diamidino-2-phenylindolee (DAPI)-stained cells and by using qPCR without PMA and reverse transcription-qPCR remained constant. The number of viable BF-1 cells in fecal samples detected by using PMA-qPCR was highly and significantly correlated with the number of viable BF-1 cells added to the fecal samples, within the range of 10 5.3 to 10 10.3 cells/g feces (wet weight) (r > 0.99, P < 0.001). After 12 healthy subjects ingested 10 10.3 to 10 11.0 CFU of BF-1 in a fermented milk product daily for 28 days, 10 4.5 ؎ 1.5 (mean ؎ standard deviation [SD]) BF-1 CFU/g was detected in fecal samples by using strain-specific selective agar; in contrast, 10 6.2 ؎ 0.4 viable BF-1 cells/g were detected by using PMA-qPCR, and a total of 10 7.6 ؎ 0.7 BF-1 cells/g were detected by using qPCR without PMA. Thus, the number of viable BF-1 cells detected by PMA-qPCR was about 50 times higher (P < 0.01) than that detected by the culture-dependent method. We conclude that strain-specific PMA-qPCR can be used to quickly and accurately evaluate viable BF-1 in feces.

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“…14,15,16,31) In particular, 0.08z BAC allowed for the extraction of intracellular ATP from yeast cells, which has proven to be quite di‹cult because of their resistant cell envelopes. 15) For simple manipulation, reagents were added in this order: 100 ml of BAC solution was added to 100 ml of test sample, followed by 100 ml of the luciferaseluciferin reagent.…”

Section: Discussionmentioning

confidence: 99%

Mutant Luciferase Enzymes from Fireflies with Increased Resistance to Benzalkonium Chloride

Hattori

Kajiyama

Maeda

et al. 2002

Bioscience, Biotechnology, and Biochemistry

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Enzymatic Treatment to Eliminate the Extracellular ATP for Improving the Detectability of Bacterial Intracellular ATP

Cited by 41 publications

References 14 publications

Structural and Functional Differences in Two Cyclic Bacteriocins with the Same Sequences Produced by Lactobacilli

Structural and Functional Differences in Two Cyclic Bacteriocins with the Same Sequences Produced by Lactobacilli

Quantitative Detection of Viable Bifidobacterium bifidum BF-1 Cells in Human Feces by Using Propidium Monoazide and Strain-Specific Primers

Mutant Luciferase Enzymes from Fireflies with Increased Resistance to Benzalkonium Chloride

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