| Determination of Polyhydroxybutyrate (PHB) Content in Ralstonia eutropha Using Gas Chromatography and Nile Red Staining
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Author:
Date:
2018-03-05
[Abstract] Ralstonia eutropha H16 produces and mobilizes (re-utilizes) intracellular polyhydroxybutyrate (PHB) granules during growth. This protocol describes the visualization of intracellular Nile red stained PHB granules and the quantification of PHB by gas chromatography. Our first method describes how to analyze PHB granules by fluorescence microscopy qualitatively. Our second approach enables the conversion of PHB to volatile hydroxycarboxylic acid methyl esters by acidic methanolysis and their quantification by gas chromatography. Through this method, it is possible to obtain an absolute quantification of PHB, e.g., per cell dry weight.
[摘要] 在生长过程中，富养罗尔斯通氏菌H16产生和动员（重新利用）细胞内聚羟基丁酸酯（PHB）颗粒。 该协议描述了细胞内尼罗红染色的PHB颗粒的可视化和通过气相色谱定量PHB。 我们的第一种方法描述了如何通过荧光显微镜定性分析PHB颗粒。 我们的第二种方法可以通过酸性甲醇分解和气相色谱定量法将PHB转化为挥发性羟基羧酸甲酯。 通过该方法，可以获得PHB的绝对定量，例如，每细胞干重。
【背景】聚羟基脂肪酸酯（PHA），尤其是聚羟基丁酸酯（PHB），是许多原核生物物种中的能量和碳储存化合物，确保细菌在压力条件下存活（Anderson和Dawes，1990;Pötter和Steinbüchel，2006; Jendrossek和Pfeiffer，2014; Bresan， >等。，2016）。这些生物聚合物的工业应用是生物降解塑料的生产（Chen，2009; Riedel等人，2015）和潜在药物成分的研究（Wu，2009; Zonari等人， ，2015; Pacheco et al。，2015; Giretova ...
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| Assays for the Detection of Rubber Oxygenase Activities
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Author:
Date:
2017-03-20
[Abstract] Microbial biodegradation of rubber relies on extracellular rubber oxygenases that catalyze the oxidative cleavage of the double bond of the polyisoprene backbone into oligo-isoprenoids. This protocol describes the determination of rubber oxygenase activities by an online measurement of molecular oxygen consumption via a non-invasive fluorescence-based assay. The produced oligo-isoprenoid cleavage products with terminal keto- and aldehyde-groups are identified qualitatively and quantitatively by HPLC. Our method allows for the characterization of homologue rubber oxygenases, and can likely be adapted to assay other oxygenases consuming dioxygen. Here we describe the determination of rubber oxygenase activities at the examples of the so far two known types of rubber oxygenases, ...
[摘要] 橡胶的微生物生物降解依赖于细胞外橡胶加氧酶，其催化聚异戊二烯骨架的双键氧化裂解成低聚异戊二烯。该方案描述了通过基于非侵入性荧光的测定在线测量分子氧消耗来测定橡胶加氧酶活性。通过HPLC定性和定量鉴定产生具有末端酮基和醛基的寡聚异戊二烯切割产物。我们的方法允许表征同系物橡胶加氧酶，并且可能适用于测定消耗双氧的其他加氧酶。在这里，我们描述了迄今为止两种已知类型的橡胶加氧酶，即橡胶加氧酶A（RoxA）和胶乳清除蛋白（Lcp）的实例的橡胶加氧酶活性的测定。
背景 ...
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| Determination of Elemental Concentrations in Lichens Using ICP-AES/MS
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Author:
Liang-Cheng Zhao, Li Wang, Yun-Jun Jiang, Yan-Qiao Hu, Chong-Ying Xu, Lei Wang, Xing Li, Li Wei, Xiu-Ping Guo, Ai-Qin Liu and Hua-Jie Liu,
Date:
2017-03-05
[Abstract] Lichens are good biomonitors for air pollution because of their high enrichment capability of atmospheric chemical elements. To monitor atmospheric element deposition using lichens, it is important to obtain information on the multi-element concentrations in lichen thalli. Because of serious air pollution, elemental concentrations in thalli of lichens from North China (especially Inner Mongolia, Hebei, Shanxi and Henan province) are often higher than those from other regions, therefore highlighting the necessity to optimize ICP-AES/MS (Inductively coupled plasma-atomic emission spectroscopy/mass spectrometry) for analyzing lichen element content. Based on the high elemental concentrations in the lichen samples, and the differences in the sensitivity and detection limits between ICP-MS and ...
[摘要] 由于大气化学元素富集能力强，地衣是空气污染的良好生物监测器。使用地衣来监测大气元素沉积，重要的是要获取有关地衣中多元素浓度的信息。由于严重的空气污染，华北地区（特别是内蒙古，河北，山西，河南等地区）的地衣单质浓度通常高于其他地区，因此强调了优化ICP-AES / MS（电感耦合等离子体原子发射光谱/质谱）用于分析地衣元素含量。基于地衣样品中高元素浓度以及ICP-MS和ICP-AES敏感性和检测限的差异，我们提出了一种使用ICP-AES / MS分析地衣31种元素的方案。二十二个元素（Cd，Ce，Co，Cr，Cs，Cu，K，La，Mo，Na，Ni，Pb，Rb，Sb，Sc，Sm，Sr，Tb，Th，Tl，V和Zn）通过使用微波消解 - ICP-MS和9种元素（Al，Ba，Ca，Fe，Mg，Mn，P，S和Ti）通过使用灰化 - 碱熔融 - ICP-AES鉴定。
背景 地衣已被广泛应用于世界许多地区的空气污染生物监测，包括在一些地区遇到重金属大气污染的中国。最近的研究发现，华北地衣的元素浓度高于其他地区（Liu等人，2016a; 2016b和2016c）的文献价值的上限，这个地区严重的空气污染。
＆nbsp;与其他技术（例如分光光度法，原子吸收光谱法和原子荧光光谱法）相比，ICP-AES / ...
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