Resazurin as an indicator of reducing capacity for analyzing the physiologic status of deep-sea bacterium Photobacterium phosphoreum ANT-2200
Resazurin as an indicator of reducing capacity for analyzing the physiologic status of deep-sea bacterium Photobacterium phosphoreum ANT-2200
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刃天青作为还原能力指标分析深海细菌磷光杆菌ANT-2200的生理状态
DOI：
10.1007/s00343-020-9301-8
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发表时间：
2020-04
影响因子：
1.6
通讯作者：
WU Long-Fei
中科院分区：
文献类型：
--
作者：
DAI Jie;TANG Hongzhi;LI Xuegong;SANTINI Claire-Lise;CUI Wenpeng;LIU Na;QI Xiaoqing;CUI Xuehua;GROSSI Alain;NOTARESCHI Philippe;ZHANG Wei-Jia;WU Long-Fei
刃天青（RZ）是一种弱荧光蓝色染料，可不可逆地还原为高荧光粉红色试卤灵（RF），RF通过光、脱氧、化学反应和细胞代谢产生的还原性有机化合物可逆地还原为无色二氢试卤灵（hRF）。由于RZ具有可靠、灵敏的荧光颜色变化和非侵入性的特点，已被广泛用作细菌、酵母和哺乳动物细胞活力/增殖测定中的氧化还原指示剂。然而，RZ很少用于海洋微生物的生理表征。在这里，我们开发了一个定制的辐射和吸收分析装置，以评估海洋细菌培养物的还原能力和生理状态。我们测量了在还原性化合物Na 2S存在下和在可见光照射下的RZ、RF和hRF的吸收光谱。在建立适当的参数后，我们监测RZ及其还原衍生物的颜色变化，以评估在各种条件下的深海细菌发光杆菌ANT-2200菌株的还原能力，生物发光和生长之间的一致性。生物发光的发射是依赖于细胞还原能力的氧化过程。随着RZ浓度的增加，ANT-2200细胞培养物的生长和生物发光逐渐受到阻碍，这表明高浓度RZ与还原代谢之间存在还原性分子的竞争。因此，在将RZ直接添加到生长培养基中以监测细胞培养物中的氧化还原反应时应谨慎。ANT-2200细胞培养物中RZ的瞬时还原速度的分析表明，高静水压力和还原能力与培养物的生物发光之间的高度一致性具有不利影响。这些数据清楚地表明了使用RZ来表征海洋细菌的微生物代谢和生理的潜力。
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 photo, deoxygenation, 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.
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影响因子：
3.7
作者：
Martini S;Al Ali B;Garel M;Nerini D;Grossi V;Pacton M;Casalot L;Cuny P;Tamburini C
通讯作者：
Tamburini C
影响因子：
2.5
作者：
Al Ali, Badr;Garel, Marc;Tamburini, Christian
通讯作者：
Tamburini, Christian
DOI：
10.1562/0031-8655(2002)076
发表时间：
2002-08
期刊：
--
影响因子：
--
作者：
J. Sutherland
通讯作者：
J. Sutherland
影响因子：
2
作者：
A. Afkhami;A. Safavi;A. Massoumi
通讯作者：
A. Afkhami;A. Safavi;A. Massoumi
影响因子：
4.5
作者：
Neumann, MG;Schmitt, CC;Bertolotti, SG
通讯作者：
Bertolotti, SG