| Determination of Hydroquinone Dioxygenase Activity
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Author:
Date:
2015-09-05
[Abstract] We recently demonstrated the presence of a quinone detoxification pathway present in Firmicutes. It is based on two enzyme activities, namely a hydroquinone dioxygenase, YaiA, described here, and a hydroquinone reductase, YaiB, described in a separate protocol. In Lactococcus lactis (L. lactis), these enzymes are encoded by the yahCD-yaiAB operon. The operon is induced by copper to prevent the synergistic toxicity of quinones and copper. The hydroquinone dioxygenase, YaiA, converts hydroquinones to 4-hydroxymuconic semialdehyde, using molecular oxygen as oxidant according to the reaction: hydroquinone + O2 → 4-hydroxymuconic semialdehyde + H+ We here describe two methods for measurements for hydroquinone dioxygenase activity, based on ...
[摘要] 我们最近展示了存在于Firmicutes中的醌解毒途径。 它基于两种酶活性，即本文所述的氢醌双加氧酶，YaiA和在单独方案中描述的氢醌还原酶，YaiB。 在乳酸乳球菌（乳酸乳杆菌）中，这些酶由yahCD-yaiAB 操纵子编码。 操纵子由铜诱导以防止醌和铜的协同毒性。 根据以下反应，氢醌双加氧酶YaiA使用分子氧作为氧化剂将氢醌转化为4-羟基粘康酸半醛：氢醌+ O 2→4-羟基粘康酸半醛+ H + 我们在这里描述了用于测量氢醌双加氧酶活性的两种方法，基于用氧电极测量的氧消耗和4-羟基粘康酸半醛的分光光度检测。 两种测定用粗细胞提取物进行。
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| Determination of Quinone Reductase Activity
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Author:
Date:
2015-09-05
[Abstract] We recently demonstrated the presence of a quinone detoxification pathway present in Firmicutes. It is based on two enzyme activities, namely a quinone reductase, YaiB, described here, and a hydroquinone dioxygenase, YaiA, described in a separate protocol. In Lactococcus lactis (L. lactis), these enzymes are encoded by the yahCD-yaiAB operon. The operon is induced by copper to prevent the synergistic toxicity of quinones and copper. The quinone reductase, YaiB, reduces p-benzoquinone and a range of quinone derivatives to hydroquinone, using NADPH as a reductant, according to the reaction: p-benzoquinone + NADPH + H+ → hydroquinone + NADP+. We here describe the measurement of quinone reductase activity, based on the ...
[摘要] 我们最近展示了存在于Firmicutes中的醌解毒途径。 它基于两种酶活性，即本文所述的醌还原酶，YaiB和在单独方案中描述的氢醌双加氧酶YaiA。 在乳酸乳球菌（乳酸乳杆菌）中，这些酶由yahCD-yaiAB 操纵子编码。 操纵子由铜诱导以防止醌和铜的协同毒性。 使用NADPH作为还原剂，醌还原酶，YaiB，根据反应，将对苯醌和一系列醌衍生物还原为氢醌：苯醌+ NADPH + H + +氢醌+ NADP + 。 我们在这里描述醌还原酶活性的测量，基于NADPH氧化的分光光度测量。
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| Measurement of Chlorophyll a and Carotenoids Concentration in Cyanobacteria
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Author:
Date:
2015-05-05
[Abstract] This is a protocol for precise measurement of chlorophyll a and total carotenoid concentrations in cyanobacteria cells. Cellular chlorophyll concentration is one of the central physiological parameters, routinely followed in many research areas ranging from stress physiology to biotechnology. Carotenoids concentration is often related to cellular stress level; combined pigments assessment provides useful insight into cellular physiological state. The current protocol was established to minimize time and equipment requirements for the routine pigments analysis. It is important to note that this protocol is suitable only for cyanobacteria containing chlorophyll a, and is not designed for species containing other chlorophyll molecules.
[摘要] 这是用于精确测量蓝细菌细胞中叶绿素a和总类胡萝卜素浓度的方案。 细胞叶绿素浓度是中心生理参数之一，在从应激生理学到生物技术的许多研究领域中常规地遵循。 类胡萝卜素浓度通常与细胞应激水平相关; 组合颜料评估提供了有用的洞察细胞生理状态。 建立了当前方案以使对常规色素分析的时间和设备要求最小化。 重要的是注意，该方案仅适用于含有叶绿素a的蓝细菌，并且不被设计用于含有其他叶绿素分子的物种。
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