合成生物学 ›› 2023, Vol. 4 ›› Issue (6): 1300-1320.DOI: 10.12211/2096-8280.2022-052

• 研究论文 • 上一篇    

希瓦氏菌Shewanella sp. MR-4在不同末端电子受体条件下的代谢通量分析

郑庆祥   

  1. 漳州人才发展集团有限公司,福建 漳州 363000
  • 收稿日期:2022-09-23 修回日期:2022-11-16 出版日期:2023-12-31 发布日期:2024-01-19
  • 通讯作者: 郑庆祥
  • 作者简介:郑庆祥(1983—),男,博士,工程师,漳州人才发展集团有限公司副总经理。研究方向为微生物代谢工程。E-mail:179949046@qq.com
  • 基金资助:
    国家自然科学基金(30970035)

Metabolic flux analysis of Shewanella sp. MR-4 with different terminal electron receptors

Qingxiang ZHENG   

  1. Zhangzhou Talent Development Group,Zhangzhou 363000,Fujian,China
  • Received:2022-09-23 Revised:2022-11-16 Online:2023-12-31 Published:2024-01-19
  • Contact: Qingxiang ZHENG

摘要:

本研究解析了Shewanella sp. MR-4在厌氧条件下以Fe3+、延胡索酸、硝酸和二甲基亚砜(DMSO)为末端电子受体时代谢通量分布,发现在各种不同末端电子受体条件下细胞内代谢通量分布发生显著改变。与延胡索酸相比,在Fe3+和硝酸为电子受体条件下,乙酸生成速率明显降低,而三羧酸循环代谢通量增加。同时,三羧酸循环回补反应存在显著差异:在延胡索酸为电子受体条件下,磷酸烯醇式丙酮酸羧化酶反应是唯一回补反应,而在Fe3+和硝酸条件下苹果酸酶催化成为唯一回补反应。本研究在代谢通量分析基础上定量解析了还原力NADH合成和消耗,分析了电子流向变化,发现在不同末端电子受体条件下,NADH产生途径和反应发生明显变化。研究结果为希瓦氏菌代谢工程改造提供了一定理论依据。

关键词: 希瓦氏菌, 代谢通量分析, 末端电子受体, 三羧酸循环, NADH

Abstract:

Shewanella is ubiquitous in natural environments, known to show various unique activities, such as metal reduction and trimethylamine production. The metal reduction capability expands the range of electron acceptors and enables its application in producing current in microbial fuel cells. Previous work primarily focused on the model organism Shewanella oneidensis MR-1, based on which the metabolic models were constructed for three other strains including Shewanella sp. MR-4. Interestingly, the genetic similarity does not always coincide with metabolic phenotype similarity between these two strains, suggesting that potential regulatory effects affecting cellular phenotypes are present in Shewanella sp.MR-4. This study analyzed the metabolic flux distribution of Shewanella sp. MR-4 when Fe3+, fumaric acid, nitric acid and dimethyl sulfoxide were used as electron acceptors under anaerobic conditions. The intracellular metabolic flux distribution was significantly changed with different electron acceptors. The formation rate of acetic acid was decreased, while the metabolic flux of the tricarboxylic acid (TCA) cycle was increased with Fe3+ and nitric acid as electron acceptors, compared to that with fumaric acid. The anaplerotic reaction in the TCA cycle was also dramatically altered: the phosphoenolpyruvate carboxylase-catalyzed reaction or malic enzyme-catalyzed reaction is the only anaplerotic reaction with fumaric acid or Fe3+ and nitric acid as electron acceptor, respectively. Based on the metabolic flux map, quantitative analysis was performed to derive the synthesis and consumption rates of reducing agent NADH, and the change of electron transfer. It was found that NADH metabolism varies significantly with the electron acceptors tested. The results provide a theoretical basis for the future metabolic engineering of Shewanella and experimental evidences that can be used for future study of different phenotypes unaccounted for in previous models of Shewanella sp. MR-4 and MR-1.

Key words: Shewanella, metabolic flux analysis, terminal electron receptor, tricarboxylic acid cycle, NADH

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