Bergmeyer Glucose Quantification for Microbiological Samples.

IF 1.2 4区 综合性期刊 Q3 MULTIDISCIPLINARY SCIENCES Jove-Journal of Visualized Experiments Pub Date : 2025-01-17 DOI:10.3791/67126
Itan Homero Ruiz-Hernandez, Luis Alberto Madrigal-Perez, Christian Alexander Aburto-Tellez, Juan Carlos González-Hernández
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Abstract

The glucose concentration is a key indicator of cellular metabolism and could indicate the total metabolism rate, an aberration in glucose metabolism, and, in some cases, how cells couple glucose metabolism to energetic metabolism. In addition, intracellular and extracellular glucose levels are indicative of the cellular metabolic status. Enzymatic techniques, such as Bergmeyer glucose quantification, are more accurate and sensitive than other techniques, such as dinitrosalicylic acid or fluorescence methods, which are usually utilized in microbiology. Although mainly used in the clinical area, Bergmeyer glucose quantification can also be applied to any cell but has not been reported in detail for bacteria, fungi, yeasts, or other microorganisms. Herein, we present a methodology to quantify glucose from bacteria and yeast samples using the enzymatic Bergmeyer glucose quantification method. The procedure involved the enzymes glucose oxidase, peroxidase, and o-dianisidine dihydrochloride incubated at 37 °C for 20 min, followed by the addition of sulfuric acid. The absorbance is then measured at 545 nm. It is important to highlight that although this technique presents difficulties in measuring high concentrations of glucose (above 60 g/L), it is possible to measure concentrations below 50 g/L using dilution factors. This enzymatic approach is valuable for research and analysis in microbiology and other scientific areas. The precision and sensitivity of the method make it helpful for detecting even low concentrations of glucose in microbiological samples.

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微生物样品的Bergmeyer葡萄糖定量。
葡萄糖浓度是细胞代谢的关键指标,可以指示总代谢率、糖代谢的畸变,在某些情况下还可以指示细胞如何将糖代谢与能量代谢耦合。此外,细胞内和细胞外的葡萄糖水平是细胞代谢状态的指示。酶技术,如Bergmeyer葡萄糖定量,比其他技术更准确和敏感,如二硝基水杨酸或荧光法,通常用于微生物学。虽然主要用于临床领域,Bergmeyer葡萄糖定量也可以应用于任何细胞,但尚未有关于细菌、真菌、酵母或其他微生物的详细报道。在这里,我们提出了一种方法来定量葡萄糖从细菌和酵母样品使用酶伯格迈耶葡萄糖定量方法。步骤包括葡萄糖氧化酶、过氧化物酶和盐酸邻二苯胺在37℃下孵育20分钟,然后加入硫酸。然后在545 nm处测量吸光度。需要强调的是,尽管该技术在测量高浓度葡萄糖(高于60 g/L)时存在困难,但使用稀释因子可以测量低于50 g/L的浓度。这种酶的方法对微生物学和其他科学领域的研究和分析是有价值的。该方法的精度和灵敏度使其有助于检测微生物样品中甚至低浓度的葡萄糖。
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来源期刊
Jove-Journal of Visualized Experiments
Jove-Journal of Visualized Experiments MULTIDISCIPLINARY SCIENCES-
CiteScore
2.10
自引率
0.00%
发文量
992
期刊介绍: JoVE, the Journal of Visualized Experiments, is the world''s first peer reviewed scientific video journal. Established in 2006, JoVE is devoted to publishing scientific research in a visual format to help researchers overcome two of the biggest challenges facing the scientific research community today; poor reproducibility and the time and labor intensive nature of learning new experimental techniques.
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