

FOLLOWUS
1.Laboratory of Deep Sea Microbial Cell Biology, Institute of Deep Sea Science and Engineering, Chinese Academy of Sciences, Sanya 572000, China
2.University of Chinese Academy of Sciences, Beijing 100049, China
3.International Associated Laboratory of Evolution and Development of Magnetotactic Multicellular Organisms (LIA-MagMC), CNRS-CAS
4.Aix Marseille University, CNRS, LCB, IMM, FR3479, IM2B, Marseille 13402, France
5.Aix Marseille University, CNRS, IMM, FR3479, Marseille 13402, France
Wei-Jia ZHANG, E-mail:wzhang@idsse.ac.cn
Long-Fei WU, E-mail:wu@imm.cnrs.fr
收稿:2019-11-13,
录用:2019-12-17,
网络首发:2020-02-12,
纸质出版:2021-01
Scan QR Code
Resazurin as an indicator of reducing capacity for analyzing the physiologic status of deep-sea bacterium
Jie DAI, Hongzhi TANG, Xuegong LI, et al. Resazurin as an indicator of reducing capacity for analyzing the physiologic status of deep-sea bacterium
Resazurin as an indicator of reducing capacity for analyzing the physiologic status of deep-sea bacterium
Jie DAI, Hongzhi TANG, Xuegong LI, et al. Resazurin as an indicator of reducing capacity for analyzing the physiologic status of deep-sea bacterium
Resazurin (RZ) is a weakly fluorescent blue dye and can be reduced irreversibly to highly fluorescent pink resorufin (RF) that is reduced reversibly to colorless dihydroresorufin (hRF) by photodeoxygenation
chemical reaction and reductive organic compounds produced through cell metabolism. Because of the reliable and sensitive fluorescence-color change and noninvasive features
RZ has been used widely as a redox indicator in cell viability/proliferation assays for bacteria
yeast
and mammalian cells. However
RZ is used rarely for physiological characterization of marine microorganisms. Here
we developed a custom-made irradiation and absorption-analysis device to assess the reducing capacity and physiologic status of marine bacterial cultures. We measured the absorption spectra of RZ
RF
and hRF in the presence of the reducing compound Na
2
S and under visible-light irradiation. After establishing appropriate parameters
we monitored the color changes of RZ and its reduced derivatives to evaluate the coherence between reducing capacity
bioluminescence and growth of the deep-sea bacterium
Photobacterium
phosphoreum
strain ANT-2200 under various conditions. Emission of bioluminescence is an oxidation process dependent upon cellular reducing capacity. Growth and bioluminescence of ANT-2200 cell cultures were impeded progressively with increasing concentrations of RZ
which suggested competition for reducing molecules between RZ at high concentration with reductive metabolism. Therefore
caution should be applied upon direct addition of RZ to growth media to monitor redox reactions in cell cultures. Analyses of the instantaneous reduction velocity of RZ in ANT-2200 cell cultures showed a detrimental effect of high hydrostatic pressure and high coherence between the reducing capacity and bioluminescence of cultures. These data clearly demonstrate the potential of using RZ to characterize the microbial metabolism and physiology of marine bacteria.
Afkhami A, Safavi A, Massoumi A. 1991. Catalytic Spectrophotometric determination of an ultra-trace amount of lead by reduction of resazurin by sodium sulfide. Analytical Letters , 24 (9): 1 643-1 655, https://doi.org/10.1080/00032719108053000..
Al Ali B, Garel M, Cuny P, Miquel J C, Toubal T, Robert A, Tamburini C. 2010. Luminous bacteria in the deep-sea waters near the ANTARES underwater neutrino telescope (Mediterranean Sea). Chemistry and Ecology , 26 (1): 57-72, https://doi.org/10.1080/02757540903513766..
Barnes S, Spenney J G. 1980. Stoichiometry of the nadh-oxidoreductase reaction for dehydrogenase determinations. Clinica Chimica Acta , 107 (3): 149-154, https://doi.org/10.1016/0009-8981(80)90442-8..
Bueno C, Villegas M L, Bertolotti S G, Previtali C M, Neumann M G, Encinas M V. 2002. The excited-state interaction of resazurin and resorufin with aminesin aqueous solutions. Photophysics and photochemical reaction. Photochemistry and Photobiology , 76 (4): 385-390, https://doi.org/10. 1562/0031-8655(2002)0760385TESIOR2.0.CO2..
De Jong D W, Woodlief W G. 1977. Fluorimetric assay of tobacco leaf dehydrogenases with resazurin. Biochimica et Biophysica Acta ( BBA ) - Enzymology , 484 (2): 249-259, https://doi.org/10.1016/0005-2744(77)90081-X..
Dunlap P V, Kita-Tsukamoto K. 2006. Luminous bacteria. In : Dworkin M, Falkow S, Rosenberg E, Schleifer K H, Stackebrandt E eds. The Prokaryotes. Volume 2: Ecophysiology and Biochemistry. Springer, New York, NY. p.863-892, https://doi.org/10.1007/0-387-30742-7_27 https://doi.org/10.1007/0-387-30742-7_27 ..
Kowaltowski A J. 2019. Strategies to detect mitochondrial oxidants. Redox Biology , 21 : 101065, https://doi.org/10. 1016/j.redox.2018.101065..
Martini S, Al Ali B, Garel M, Nerini D, Grossi V, Pacton M, Casalot L, Cuny P, Tamburini C. 2013. Effects of hydrostatic pressure on growth and luminescence of a moderately-piezophilic luminous bacteria Photobacterium phosphoreum ANT-2200. PLoS One , 8 (6): e66580, https://doi.org/10.1371/journal.pone.0066580..
Neumann M G, Schmitt C C, Previtali C M, Bertolotti S G. 1996. Photoreduction of resazurin in the presence of aliphatic amines. Dyes and Pigments , 32 (2): 93-99, https://doi.org/10.1016/0143-7208(96)00020-4..
Nijvipakul S, Wongratana J, Suadee C, Entsch B, Ballou D P, Chaiyen P. 2008. LuxG is a functioning flavin reductase for bacterial luminescence. Journal of Bacteriology , 190 (5): 1 531-1 538, https://doi.org/10.1128/JB.01660-07..
O'Brien J, Wilson I, Orton T, Pognan F. 2000. Investigation of the Alamar Blue (resazurin) fluorescent dye for the assessment of mammalian cell cytotoxicity. European Journal of Biochemistry , 267 (17): 5 421-5 426, https:// doi.org/10.1046/j.1432-1327.2000.01606.x..
Porcal G V, Altamirano M S, Bertolotti S G, Previtali C M. 2011. Organized media effect on the photochemical deoxygenation of resazurin in the presence of triethanolamine. Journal of Photochemistry and Photobiology A : Chemistry , 219 (1): 62-66, https://doi.org/10.1016/j.jphotochem.2011.01.018..
Rampersad S N. 2012. Multiple applications of Alamar Blue as an indicator of metabolic function and cellular health in cell viability bioassays. Sensors , 12 (9): 12 347-12 360, https://doi.org/10.3390/s120912347..
Safavi A, Afkhami A, Massoumi A. 1990. Spectrophotometric catalytic determination of ultra-trace amounts of selenium based on the reduction of resazurin by sulphide. Analytica Chimica Acta , 232 : 351-356, https://doi.org/10.1016/S0003-2670(00)81253-2..
Zhang S D, Santini C L, Zhang W J, Barbe V, Mangenot S, Guyomar C, Garel M, Chen H T, Li X G, Yin Q J, Zhao Y, Armengaud J, Gaillard J C, Martini S, Pradel N, Vidaud C, Alberto F, Médigue C, Tamburini C, Wu L F. 2016. Genomic and physiological analysis reveals versatile metabolic capacity of deep-sea Photobacterium phosphoreum ANT-2200. Extremophiles , 20 (3): 301-310, https://doi.org/10.1007/s00792-016-0822-1..
Zhao B Z, Ranguelova K, Jiang J J, Mason R P. 2011. Studies on the photosensitized reduction of resorufin and implications for the detection of oxidative stress with Amplex Red. Free Radical Biology and Medicine , 51 (1): 153-159, https://doi.org/10.1016/j.freeradbiomed.2011.03.016..
0
浏览量
0
Downloads
0
CSCD
关联资源
相关文章
相关作者
相关机构

京公网安备11010802024621