HPLC–UV method for simultaneous determination of adenosine triphosphate and its metabolites in Mycobacterium smegmatis

• Autorzy: Akhova Anna V. , Tkachenko Alexander G.

Identyfikatory

DOI

10.1556/1326.2017.00344

• Języki publikacji: EN

Abstrakty

EN

Method for the analysis of intracellular adenosine triphosphate (ATP), adenosine diphosphate (ADP), and adenosine monophosphate (AMP) in Mycobacterium smegmatis that involves rapid extraction procedure based on sonication of cells in perchloric acid, as well as separation of nucleotides by ion-pair reversed-phase high-performance liquid chromatography and ultraviolet (UV) detection at 254 nm, is developed. The analytes were separated with mobile phase consisted of acetonitrile and 50 mM monobasic potassium phosphate (pH 4.6) with 25 mM tetrabutylammonium hydrogensulfate in a ratio of 0.5:99.5% within 30 min. The calibration curves were linear in the range of 20–1000 pmol of ATP and 10–1000 pmol of ADP and AMP with correlation coefficient (r2) of ≥0.9998. The proposed method is applicable for mycobacterium cultures taken over a wide range of optical density and physiological states. Concentrations of ATP, ADP, and AMP in mycobacterial extracts varied from 2.61 ± 0.27 to 9.60 ± 0.19 nmol/mg dry weight, from 1.75 ± 0.12 to 5.86 ± 0.09 nmol/mg dry weight, and from 0.55 ± 0.08 to 4.40 ± 0.07 nmol/mg dry weight, respectively, depending on the physiological state.

Słowa kluczowe

EN

Adenine nucleotides; ATP; energy charge; HPLC; mycobacterium

• Wydawca: University of Silesia in Katowice ; Akademiai Kiado
• Czasopismo: Acta Chromatographica
• Rocznik: 2019
• Tom: Vol. 31, no. 1
• Strony: 45--48
• Opis fizyczny: Bibliogr. 23 poz., rys.

Twórcy

autor

Akhova Anna V.

• Institute of Ecology and Genetics of Microorganisms, Russian Academy of Sciences, 13, Golev str., Perm 614081, Russia
• Perm National Research Polytechnic University, 29, Komsomolsky Ave., Perm 614990, Russia

autor

Tkachenko Alexander G.

• Institute of Ecology and Genetics of Microorganisms, Russian Academy of Sciences, 13, Golev str., Perm 614081, Russia
• Perm State National Research University, 15, Bukirev str., Perm 614068, Russia

Bibliografia

• [1] Atkinson, D.; Walton, G. J. Biol. Chem. 1967, 242, 3239.
• [2] Davis, W. M.; White, D. C. Appl. Env. Microb. 1980, 40, 539.
• [3] Abelho, M. In Methods to Study Litter Decomposition: A Practical Guide; Graca, M. A. S, Barlocher, F., Gessner, M. O., Eds.; Springer: Netherlands, 2005; pp. 223–230.
• [4] Chapman, A. G.; Fall, L.; Atkinson, D. E. J. Bacteriol. 1971, 108, 1072.
• [5] Lee, Y. N.; Colston, M. J. FEMS Microb. Lett. 1986, 35, 279.
• [6] Franzblau, S. G.; DeGroote, M. A.; Cho, S. H.; Andries, K.; Nuermberger, E.; Orme, I. M.; Mdluli, K.; Angulo-Barturen, I.; Dick, T.; Dartois, V.; Lenaerts, A. J. Tuberculosis 2012, 92, 453.
• [7] Bald, D.; Villellas, C.; Lu, P.; Koul, A. mBio 2017, 8, e00272-17.
• [8] Lundin, A.; Thore, A. Appl. Microb. 1975, 30, 713.
• [9] Werner, A.; Schneider, W.; Siems, W.; Grune, T.; Schreiter, C. Chromatographia 1989, 27, 639.
• [10] Brugidou, C.; Rocher, A.; Giraud, E.; Lelong, B.; Marin, B.; Raimbault, M. Biotechnol. Tech. 1991, 5, 475.
• [11] Muller, R. H.; Loffhagen, N.; Babel, W. J. Microb. Meth. 1996, 25, 29.
• [12] Feng, H.T.; Wong, N.; Wee, S.; Lee, M. M. J. Chromatogr. B 2008, 870, 131.
• [13] Tomiya, N.; Ailor, E.; Lawrence, S. M.; Betenbaugh, M. J.; Lee, Y. C. Anal. Biochem. 2001, 293, 129.
• [14] Perrett, D. Chromatographia 1982, 16, 211.
• [15] Tkachenko, A. G.; Chudinov, A. A. Prikl. Biokhim. Mikrobiol. 1985, 21, 553.
• [16] Schneider, D. A.; Gourse, R. L. J. Biol. Chem. 2004, 279, 8262.
• [17] Maglica, Ž.; Özdemir, E.; McKinney, J. D. mBio 2015, 6, e02236-14.
• [18] Cavallini, A.; De Micheli, G.; Carrara, S. Sensor. Lett. 2011, 9, 1.
• [19] Zhu, P.; Wang, J.; Wang, L.; Zhou, L.; Shi, P. Acta Chromatogr. 2016, 28, 501.
• [20] Kehr, J.; Chavko, M. Z. Anal. Chem. 1986, 325, 466.
• [21] Khlyntseva, S. V.; Bazel, Y. R.; Vishnikin, A. B.; Andruch, V. J. Anal. Chem. 2009, 64, 657.
• [22] Helenius, M.; Jalkanen, S.; Yegutkin, G. G. Biochim. Biophys. Acta 2012, 1823, 1967.
• [23] Dhople, A. M.; Hanks, J. H. Appl. Microb. 1973, 26, 399.

Uwagi

PL

Opracowanie rekordu w ramach umowy 509/P-DUN/2018 ze środków MNiSW przeznaczonych na działalność upowszechniającą naukę (2019).