Kim Jong-Geol, Park Soo-Je, Sinninghe Damsté Jaap S, Schouten Stefan, Rijpstra W Irene C, Jung Man-Young, Kim So-Jeong, Gwak Joo-Han, Hong Heeji, Si Ok-Ja, Lee SangHoon, Madsen Eugene L, Rhee Sung-Keun
Department of Microbiology, Chungbuk National University, Cheongju 361-763, South Korea;
Department of Biology, Jeju National University, Jeju 690-756, South Korea;
Proc Natl Acad Sci U S A. 2016 Jul 12;113(28):7888-93. doi: 10.1073/pnas.1605501113. Epub 2016 Jun 23.
Ammonia-oxidizing archaea (AOA), that is, members of the Thaumarchaeota phylum, occur ubiquitously in the environment and are of major significance for global nitrogen cycling. However, controls on cell growth and organic carbon assimilation by AOA are poorly understood. We isolated an ammonia-oxidizing archaeon (designated strain DDS1) from seawater and used this organism to study the physiology of ammonia oxidation. These findings were confirmed using four additional Thaumarchaeota strains from both marine and terrestrial habitats. Ammonia oxidation by strain DDS1 was enhanced in coculture with other bacteria, as well as in artificial seawater media supplemented with α-keto acids (e.g., pyruvate, oxaloacetate). α-Keto acid-enhanced activity of AOA has previously been interpreted as evidence of mixotrophy. However, assays for heterotrophic growth indicated that incorporation of pyruvate into archaeal membrane lipids was negligible. Lipid carbon atoms were, instead, derived from dissolved inorganic carbon, indicating strict autotrophic growth. α-Keto acids spontaneously detoxify H2O2 via a nonenzymatic decarboxylation reaction, suggesting a role of α-keto acids as H2O2 scavengers. Indeed, agents that also scavenge H2O2, such as dimethylthiourea and catalase, replaced the α-keto acid requirement, enhancing growth of strain DDS1. In fact, in the absence of α-keto acids, strain DDS1 and other AOA isolates were shown to endogenously produce H2O2 (up to ∼4.5 μM), which was inhibitory to growth. Genomic analyses indicated catalase genes are largely absent in the AOA. Our results indicate that AOA broadly feature strict autotrophic nutrition and implicate H2O2 as an important factor determining the activity, evolution, and community ecology of AOA ecotypes.
氨氧化古菌(AOA),即奇古菌门的成员,在环境中普遍存在,对全球氮循环具有重要意义。然而,人们对AOA的细胞生长和有机碳同化的控制了解甚少。我们从海水中分离出一株氨氧化古菌(命名为菌株DDS1),并利用该菌株研究氨氧化的生理学。使用另外四株来自海洋和陆地生境的奇古菌菌株证实了这些发现。与其他细菌共培养时,以及在添加了α-酮酸(如丙酮酸、草酰乙酸)的人工海水培养基中,菌株DDS1的氨氧化作用增强。AOA的α-酮酸增强活性以前被解释为兼性营养的证据。然而,异养生长试验表明,丙酮酸掺入古菌膜脂的量可以忽略不计。相反,脂质碳原子来自溶解的无机碳,表明其严格的自养生长。α-酮酸通过非酶促脱羧反应自发地清除过氧化氢,这表明α-酮酸作为过氧化氢清除剂的作用。事实上,其他能清除过氧化氢的试剂,如二甲基硫脲和过氧化氢酶,取代了对α-酮酸的需求,促进了菌株DDS1的生长。实际上,在没有α-酮酸的情况下,菌株DDS1和其他AOA分离株被证明会内源性产生过氧化氢(高达约4.5μM),这对生长具有抑制作用。基因组分析表明,AOA中基本上不存在过氧化氢酶基因。我们的结果表明,AOA广泛具有严格的自养营养特征,并表明过氧化氢是决定AOA生态型的活性、进化和群落生态学的重要因素。