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毛细管电泳-质谱联用技术揭示了 Synechocystis sp. PCC 6803 在氮饥饿期间碳代谢物的分布。

Capillary electrophoresis-mass spectrometry reveals the distribution of carbon metabolites during nitrogen starvation in Synechocystis sp. PCC 6803.

机构信息

RIKEN Plant Science Center, 1-7-22 Suehiro-cho, Tsurumi-ku, Yokohama, Kanagawa, 230-0045, Japan; PRESTO, Japan Science and Technology Agency (JST), 4-1-8 Honcho, Kawaguchi, Saitama, 332-0012, Japan.

出版信息

Environ Microbiol. 2014 Feb;16(2):512-24. doi: 10.1111/1462-2920.12170. Epub 2013 Jun 25.

Abstract

Nitrogen availability is one of the most important factors for the survival of cyanobacteria. Previous studies on Synechocystis revealed a contradictory situation with regard to metabolism during nitrogen starvation; that is, glycogen accumulated even though the expressions of sugar catabolic genes were widely upregulated. Here, we conducted transcript and metabolomic analyses using capillary electrophoresis-mass spectrometry on Synechocystis sp. PCC 6803 under nitrogen starvation. The levels of some tricarboxylic acid cycle intermediates (succinate, malate and fumarate) were greatly increased by nitrogen deprivation. Purine and pyrimidine nucleotides were markedly downregulated under nitrogen depletion. The levels of 19 amino acids changed under nitrogen deprivation, especially those of amino acids synthesized from pyruvate and phosphoenolpyruvate, which showed marked increases. Liquid chromatography-mass spectrometry analysis demonstrated that the amount of NADPH and the NADPH/NADH ratio decreased under nitrogen depletion. These data demonstrate that there are increases in not only glycogen but also in metabolites downstream of sugar catabolism in Synechocystis sp. PCC 6803 under nitrogen starvation, resolving the contradiction between glycogen accumulation and induction of sugar catabolic gene expression in this unicellular cyanobacterium.

摘要

氮素供应是蓝藻生存的最重要因素之一。先前对集胞藻 PCC 6803 的研究表明,在氮饥饿期间,其代谢存在矛盾情况,即尽管糖分解代谢基因广泛上调,但仍会积累糖原。在此,我们对氮饥饿条件下的集胞藻 PCC 6803 进行了毛细管电泳-质谱联用的转录组和代谢组学分析。氮饥饿大大增加了某些三羧酸循环中间产物(琥珀酸、苹果酸和富马酸)的水平。嘌呤和嘧啶核苷酸在氮耗尽时明显下调。氮饥饿下 19 种氨基酸的水平发生变化,特别是那些由丙酮酸和磷酸烯醇丙酮酸合成的氨基酸,其水平显著增加。液相色谱-质谱分析表明,氮饥饿下 NADPH 的量和 NADPH/NADH 比值降低。这些数据表明,在氮饥饿条件下,集胞藻 PCC 6803 不仅糖原增加,而且糖分解代谢下游的代谢物也增加,解决了该单细胞蓝藻中糖原积累和糖分解代谢基因表达诱导之间的矛盾。

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