| Pipecolic Acid Quantification Using Gas Chromatography-coupled Mass Spectrometry
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Author:
Date:
2020-12-05
[Abstract] Pipecolic acid (Pip), a non-proteinacious product of lysine catabolism, is an important regulator of immunity in plants and humans alike. For instance, Pip accumulation is associated with the genetic disorder Zellweger syndrome, chronic liver diseases, and pyridoxine-dependent epilepsy in humans. In plants, Pip accumulates upon pathogen infection and is required for plant defense. The aminotransferase ALD1 catalyzes biosynthesis of Pip precursor piperideine-2-carboxylic acid, which is converted to Pip via ornithine cyclodeaminase. A variety of methods are used to quantify Pip, and some of these involve use of expensive amino acid analysis kits. Here, we describe a simplified procedure for quantitative analysis of Pip from plant tissues. Pipecolic acid was extracted from leaf tissues along ...
[摘要] [摘要]胡椒酸（Pip）是赖氨酸分解代谢的非蛋白质产物，是植物和人类免疫力的重要调节剂。例如，Pip积累与人类遗传病Zellweger综合征，慢性肝病和吡ido醇依赖性癫痫有关。在植物中，Pip会在病原体感染时积聚，是植物防御所必需的。氨基转移酶ALD1催化Pip前体哌啶-2-羧酸的生物合成，后者通过鸟氨酸环脱氨酶转化为Pip。多种方法可用于定量Pip，其中一些方法涉及使用昂贵的氨基酸分析试剂盒。在这里，我们描述了从植物组织中定量定量分析的简化程序。P ipecolic酸是从叶组织中提取与内标正缬氨酸一起衍生与丙基氯甲酸酯和分析通过气相色谱-质谱联用选择性离子模式。此过程简单，经济且有效，并且不涉及同位素内标或多步衍生化。
关键词：胡椒酸GC-MS拟南芥植物防御氯甲酸丙酯
[背景]匹，非蛋白氨基酸赖氨酸的氨基酸衍生物，积聚在病原体感染的植物的本地和远端的组织和患者的体液与过氧化物酶体疾病（Schutgens等人，1986;尹和An，2010;Návarová等人。，2012;王等人，2018） ...
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| Incubation of Cyanobacteria under Dark, Anaerobic Conditions and Quantification of the Excreted Organic Acids by HPLC
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Author:
Date:
2017-05-05
[Abstract] Succinate and lactate are commodity chemicals used for producing bioplastics. Recently, it was found that such organic acids are excreted from cells of the unicellular cyanobacterium Synechocystis sp. PCC 6803 under dark, anaerobic conditions. To conduct the dark, anaerobic incubation, cells were concentrated within a vial that was then sealed with a butyl rubber cap, following which N2 gas was introduced into the vial. The organic acids produced were quantified by high-performance liquid chromatography via post-labeling with bromothymol blue as a pH indicator. After separation by ion-exclusion chromatography, the organic acids were identified by comparing their retention time with that of standard solutions. These procedures allow researchers to quantify the organic ...
[摘要] 琥珀酸盐和乳酸盐是用于生产生物塑料的商品化学品。最近发现这样的有机酸从单细胞蓝细菌集胞藻的细胞中排出。 PCC 6803在黑暗，无氧条件下。为了进行黑暗的厌氧培养，将细胞浓缩在小瓶内，然后用丁基橡胶帽密封，随后将N 2气体引入小瓶中。通过用溴百里酚蓝作为pH指示剂的后标记，通过高效液相色谱定量产生的有机酸。通过离子排阻色谱分离后，通过将其保留时间与标准溶液的保留时间进行比较来鉴定有机酸。这些程序允许研究人员量化微生物产生的有机酸，有助于蓝藻生物学和生物技术的知识。
背景 蓝细菌是一组进行氧合光合作用的细菌。集胞藻 sp。 PCC 6803（以下称为集胞藻6803）是非固氮，单细胞蓝细菌，通常用于基础和应用研究。集胞藻6803细胞能够在黑暗和厌氧条件下分泌有机酸，如琥珀酸盐和乳酸盐（Osanai等，2015）。培养条件的遗传操作和修饰，例如添加钾或NaHCO 3，成功地提高了从集胞藻6803细胞排出的有机酸的水平（Osanai >等人，2015; Hasunuma等人，2016; Iijima等人，2016; Ueda等人。 ...
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