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Comparison of Methods for Extraction of Bacterial Adenine Nucleotides Determined by Firefly Assay
Open accessAbstract
Adenine nucleotides in Escherichia coli, Bacillus cereus, Klebsiella pneumoniae, Staphylococcus aureus, and Pseudomonas aeruginosa were extracted using 10 different methods. Extracts were assayed for adenosine 5'-triphosphate (ATP), adenosine 5'-diphosphate (ADP), and adenosine 5'-monophosphate (AMP) by the firefly method using an improved procedure. Analytical interference by bacterial enzymes not inactivated during the extraction was found to be a major problem. However, these enzymes were inactivated to a considerable extent by the inclusion of ethylenediaminetetraacetate in the extraction reagent. The 10 extraction methods were compared with respect to yield of adenine nucleotides, interference with the enzymic assay, reproducibility of the method, and stability of the extracts. Results indicated that extraction with trichloroacetic acid was the method most closely reflecting actual levels of ATP, ADP and AMP in intact bacterial cells. However, for the extraction of ATP in some bacterial strains several other methods may be used and may be advantageous from a practical point of view.
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Citations
Cited by
Use of firefly luciferase in atp-related assays of biomass, enzymes, and metabolites
Lundin, A. (2000) · Methods in Enzymology · doi:10.1016/s0076-6879(00)05499-9 · Cited by 272
Self-citationA Review of Methods to Determine Viability, Vitality, and Metabolic Rates in Microbiology
Braissant, O., Astasov-Frauenhoffer, M., Waltimo, T., & Bonkat, G. (2020) · Frontiers in Microbiology · doi:10.3389/fmicb.2020.547458 · Cited by 201
Varik, V., Oliveira, S. R. A., Hauryliuk, V., & Tenson, T. (2017) · Scientific Reports · doi:10.1038/s41598-017-10988-6 · Cited by 124
Glucose triggers ATP secretion from bacteria in a growth-phase-dependent manner
Hironaka, I., Iwase, T., Sugimoto, S., Okuda, K.-I., Tajima, A., Yanaga, K., & Mizunoe, Y. (2013) · Applied and Environmental Microbiology · doi:10.1128/aem.03871-12 · Cited by 87
McMillan, D. G. G., Ferguson, S. A., Dey, D., Schröder, K., Aung, H. L., Carbone, V., Attwood, G. T., Ronimus, R. S., Meier, T., Janssen, P. H., et al. (2011) · Journal of Biological Chemistry · doi:10.1074/jbc.m111.281675 · Cited by 42
Metadata
- Authors
- Lundin, A., & Thore, A.
- Venue
- Applied Microbiology
- Publisher
- American Society for Microbiology
- Published
- 1975-01-01
- Volume
- 30
- Issue
- 5
- Pages
- 713-721
- ISSN
- 0003-6919 (ISSN-L)
- OpenAlex
- W1493193010
- Cited by
- 299
- References
- 0
- FWCI
- 4.05
- Type
- journal article
- Language
- en
- Open access
- diamond
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- Lundin, A., & Thore, A. (1975). Comparison of Methods for Extraction of Bacterial Adenine Nucleotides Determined by Firefly Assay. Applied Microbiology. https://doi.org/10.1128/aem.30.5.713-721.1975
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Contains information from the Semantic Scholar Academic Graph Datasets, made available under the ODC Attribution License (ODC-BY 1.0).
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