极端嗜酸热古菌Acidianus manzaensis硫氧化模型构建

为了解极端嗜酸热古菌硫氧化代谢途径,基于Acidianus manzaensis YN-25全基因组信息,通过NCBI数据库比对初步筛选了20个可能与硫氧化相关的基因,并通过实时定量PCR(RT-qPCR)比较了所筛选基因在以单质硫(S~0)和亚铁(Fe2+)两种不同能源底物培养下的表达差异。结果表明,A.manzaensis菌中存在至少15个与硫氧化相关的基因,经过比对分析,它们包括5个编码氧化单质硫(S~0)及含硫中间产物的酶基因、4个编码末端氧化酶基因、1个编码硫酸根转运蛋白酶基因、1个编码电子传递蛋白基因,以及4个编码与硫氧化密切相关的硫还原蛋白家族(Sulfur reduction protein)的dsr E基因。基于上述实验分析结果,拟构建极端嗜酸热古菌A.manzaensis的硫氧化模型,即胞外的S~0跨膜转运进入细胞内后,经硫氧化蛋白(SOR)氧化还原生成S_2O_3~(2-),SO_3~(2-)和H_2S等含硫中间产物。接着,细胞内通过其他相关硫氧化酶的作用将这些中间产物进一步氧化,并将氧化得到的电子传递给细胞膜上的氧化型醌(Q~(2+)),使其形成还原型的醌(QH_2),QH_2最终被末端氧化酶氧化形成NADH和ATP,从而为细胞生长提供能量。     

超嗜热古菌整合性遗传元件的研究进展

细菌中整合性遗传元件与DNA修饰和防御、毒力因子传播以及次级代谢等生理功能存在关联,而相关研究在超嗜热古菌中尚处于起步阶段。本文综述了超嗜热古菌中整合性病毒、质粒及基因组岛等整合性遗传元件的分类、整合及维持机制。展示了整合性遗传元件参与的水平基因转移过程在超嗜热古菌基因组演化中扮演的重要角色。整合性遗传元件相关功能基因组学研究为理解超嗜热古菌的多样性及其环境适应性机制提供了新的视角。     

极端嗜热古菌DNA修复核酸内切酶的研究进展

极端嗜热古菌由于生活在高温环境,其基因组DNA面临着严重的挑战,因此,它们如何维持其基因组稳定是本研究领域最为关注的科学问题之一。极端嗜热古菌具有与常温微生物相似的自发突变频率,暗示着它们比常温微生物具有更加有效的DNA修复体系进行修复高温所造成的基因组DNA损伤。目前,极端嗜热古菌DNA修复的分子机制尚不清楚。核酸内切酶在DNA修复途径中发挥着重要的作用。基因组序列显示极端嗜热古菌编码多种DNA修复核酸内切酶,但是其研究尚处于初期阶段。本文综述了极端嗜热古菌DNA修复核酸内切酶Nuc S、Endo V、Endo Q、XPF和Hjc的研究进展,并对今后的研究提出了展望。     

极端嗜盐古菌蛋白类抗生素——嗜盐菌素

古菌(Archaea)是一类与细菌及真核生物显著不同的生命的第三种形式[1],大多生活在极端或特殊环境,主要包括产甲烷古菌(Methanogenic Achaea)、极端嗜盐古菌(Extremely Halophilic Archaea)和极端嗜热古菌(Extremely Thermophilic Archaea)等三大类.极端古菌是极端环境微生物的重要成员,也是极端环境微生物资源开发的重要领域.其中,嗜盐古菌可产生一类蛋白类抗生素,称为嗜盐菌素(halocin).     

陆地生态系统甲烷产生和氧化过程的微生物机理

陆地生态系统存在许多常年性或季节性缺氧环境,如:湿地、水稻土、湖泊沉积物、动物瘤胃、垃圾填埋场和厌氧生物反应器等。每年有大量有机物质进入这些环境,在缺氧条件下发生厌氧分解。甲烷是有机质厌氧分解的最终产物。产生的甲烷气体可通过缺氧-有氧界面释放到大气,产生温室效应,是重要的温室气体。产甲烷过程是缺氧环境中有机质分解的核心环节,而甲烷氧化是缺氧-有氧界面的重要微生物过程。甲烷的产生和氧化过程共同调控大气甲烷浓度,是全球碳循环不可分割的组成部分。对陆地生态系统甲烷产生和氧化过程的微生物机理研究进展进行了概要回顾和综述。主要内容包括:新型产甲烷古菌即第六和第七目产甲烷古菌和嗜冷嗜酸产甲烷古菌的发现;短链脂肪酸中间产物互营氧化过程与直接种间电子传递机制;新型甲烷氧化菌包括厌氧甲烷氧化菌和疣微菌属好氧甲烷氧化菌的发现;甲烷氧化菌生理生态与环境适应的新机制。这些研究进展显著拓展了人们对陆地生态系统甲烷产生和氧化机理的认识和理解。随着新一代土壤微生物研究技术的发展与应用,甲烷产生和氧化微生物研究领域将面临更多机遇和挑战,对未来发展趋势做了展望。     

湖泊硫循环微生物研究进展

湖泊是响应气候和环境变化的关键生态系统,是研究元素(如碳、氮和硫等)生物地球化学循环的热点环境。湖泊(尤其咸盐湖)具有硫酸盐含量高且含硫化合物种类丰富的特点,因而湖泊中硫元素生物地球化学循环过程非常活跃。微生物是驱动湖泊硫循环的重要推手。因此,研究湖泊中微生物参与的硫元素生物地球化学循环过程以及相关微生物类群构成,对于深入探索微生物在湖泊生态系统中的作用具有重要意义。本文综述了湖泊中驱动硫循环的微生物(硫氧化菌和硫酸盐还原菌)种群多样性、功能基因、代谢途径、硫氧化/硫酸盐还原速率及其对环境条件变化响应等方面的研究现状,并对未来湖泊微生物驱动的硫循环研究方向进行了展望。     

极端嗜盐古菌蛋白类抗生素——嗜盐菌素

古菌 (Ａｒｃｈａｅａ)是一类与细菌及真核生物显著不同的生命的第三种形式[1] ,大多生活在极端或特殊环境 ,主要包括产甲烷古菌 (ＭｅｔｈａｎｏｇｅｎｉｃＡｃｈａｅａ)、极端嗜盐古菌 (ＥｘｔｒｅｍｅｌｙＨａｌｏｐｈｉｌｉｃＡｒｃｈａｅａ)和极端嗜热古菌 (ＥｘｔｒｅｍｅｌｙＴｈｅｒｍｏｐｈｉｌｉｃＡｒｃｈａｅａ)等三大类。极端古菌是极端环境微生物的重要成员 ,也是极端环境微生物资源开发的重要领域。其中 ,嗜盐古菌可产生一类蛋白类抗生素 ,称为嗜盐菌素 (ｈａｌｏｃｉｎ)。与细菌素相似[2 ] ,嗜盐菌素是由质粒编码、核糖体合…     

嗜盐古菌分类学研究进展

嗜盐古菌是一类需要高盐维持生长的古菌。到目前为止,已发现的嗜盐古菌都属于古菌域的广古菌门,主要包括:嗜盐甲烷古菌类群、嗜盐古菌纲的全部成员以及尚不能培养的纳米嗜盐古菌类群。嗜盐古菌是盐环境的土著类群,驱动着盐环境生态系统的生物地球化学循环。作为极端微生物,嗜盐古菌在理论研究和应用领域具有重要的研究价值。本文从嗜盐古菌分类学地位的变迁、分类学方法、分类学研究现状及我国的嗜盐古菌分类学研究等方面综述了嗜盐古菌分类学的最新研究进展。     

甲烷氧化菌及其在环境治理中的应用

甲烷的生物氧化包括好氧氧化和厌氧氧化两种,分别由好氧甲烷氧化菌和厌氧甲烷氧化菌完成.由于该过程是减少自然环境中温室气体甲烷排放的重要途径,越来越受到各国学者的重视.本文主要对当前甲烷氧化菌的研究现状进行了综述,对好氧甲烷氧化菌的种类、参与氧化甲烷的关键酶,厌氧甲烷氧化菌的种类、参与的微生物菌种以及氧化机理进行了论述,并对这两类微生物在温室气体减排、污染物治理、废水生物脱氮、硫及金属元素回收等方面的应用现状及前景进行了分析.     

超嗜热古菌遗传操作系统研究进展

摘要：遗传操作系统,是研究基因和基因产物功能的一个极为重要的工具。超嗜热古菌遗传操作系统方面的研究落后于甲烷菌及嗜盐古菌中的研究,主要原因是选择标记的缺乏。然而,近十年来,在以硫化叶菌(Sulfolobus)为代表的超嗜热泉古菌和Thermococcus kodakaraensis为代表的超嗜热广古菌中,遗传操作系统研究取得了很大的进展。本文主要对这两种超嗜热古菌的遗传操作系统进展以及应用进行概述。     

Sulfur metabolism in archaea reveals novel processes

Studies on sulfur metabolism in archaea have revealed many novel enzymes and pathways and have advanced our understanding on metabolic processes, not only of the archaea, but of biology in general. A variety of dissimilatory sulfur metabolisms, i.e. reactions used for energy conservation, are found in archaea from both the Crenarchaeota and Euryarchaeota phyla. Although not yet fully characterized, major processes include aerobic elemental sulfur (S(0) ) oxidation, anaerobic S(0) reduction, anaerobic sulfate/sulfite reduction and anaerobic respiration of organic sulfur. Assimilatory sulfur metabolism, i.e. reactions used for biosynthesis of sulfur-containing compounds, also possesses some novel features. Cysteine biosynthesis in some archaea uses a unique tRNA-dependent pathway. Fe-S cluster biogenesis in many archaea differs from that in bacteria and eukaryotes and requires unidentified components. The eukaryotic ubiquitin system is conserved in archaea and involved in both protein degradation and biosynthesis of sulfur-containing cofactors. Lastly, specific pathways are utilized for the biosynthesis of coenzyme M and coenzyme B, the sulfur-containing cofactors required for methanogenesis.     

Dissimilatory Oxidation and Reduction of Elemental Sulfur in Thermophilic Archaea

The oxidation and reduction of elemental sulfur and reduced inorganic sulfur species are some of the most important energy-yielding reactions for microorganisms living in volcanic hot springs, solfataras, and submarine hydrothermal vents, including both heterotrophic, mixotrophic, and chemolithoautotrophic, carbon dioxide-fixing species. Elemental sulfur is the electron donor in aerobic archaea like Acidianus and Sulfolobus. It is oxidized via sulfite and thiosulfate in a pathway involving both soluble and membrane-bound enzymes. This pathway was recently found to be coupled to the aerobic respiratory chain, eliciting a link between sulfur oxidation and oxygen reduction at the level of the respiratory heme copper oxidase. In contrast, elemental sulfur is the electron acceptor in a short electron transport chain consisting of a membrane-bound hydrogenase and a sulfur reductase in (facultatively) anaerobic chemolithotrophic archaea Acidianus and Pyrodictium species. It is also the electron acceptor in organoheterotrophic anaerobic species like Pyrococcus and Thermococcus, however, an electron transport chain has not been described as yet. The current knowledge on the composition and properties of the aerobic and anaerobic pathways of dissimilatory elemental sulfur metabolism in thermophilic archaea is summarized in this contribution.     

微生物硫代谢及其驱动下建立的生物生态关系

硫在环境中广泛存在,是生物细胞的主要构成元素,微生物、动物和植物的硫基础代谢途径之间存在着广泛联系.本文以微生物硫代谢为主线,全面总结了硫在3类生物中的4条主要代谢途径,并重点阐明了其共性、区别及联系.微生物参与了所有硫的主要代谢,是驱动硫生物循环的主要动力.微生物异化硫还原降低了环境中甲烷的挥发,微生物、植物实施的同...     

Detection of Metabolic Key Enzymes of Methane Turnover Processes in Cold Seep Microbial Biofilms

In anoxic environments, methane oxidation is conducted in a syntrophic process between methanotrophic archaea (ANME) and sulfate reducing bacteria (SRB). Microbial mats consisting of ANME, SRB and other microorganisms form methane seep-related carbonate buildups in the anoxic bottom waters of the Black Sea Crimean shelf. To shed light on the localization of the biochemical processes at the level of single cells in the Black Sea microbial mats, we applied antibody-based markers for key enzymes of the relevant metabolic pathways. The dissimilatory adenosine-5′-phosphosulfate (APS) reductase, methyl-coenzyme M reductase (MCR) and methanol dehydrogenase (MDH) were selected to localize sulfate respiration, reverse methanogenesis and aerobic methane oxidation, respectively. The key enzymes could be localized by double immunofluorescence and immunocytochemistry at light- and electron microscopic levels. In this study we show that sulfate reduction is conducted synchronized and in direct proximity to reverse methanogenesis of ANME archaea. Microcolonies in interspaces between ANME/SRB express methanol dehydrogenase, which is indicative for oxidation of C₁ compounds by methylotrophic or methanotrophic bacteria. Thus, in addition to syntrophic AOM, oxygen-dependent processes are also conducted by a small proportion of the microbial population.     

Sulfur oxidizers dominate carbon fixation at a biogeochemical hot spot in the dark ocean

Bacteria and archaea in the dark ocean (>200 m) comprise 0.3–1.3 billion tons of actively cycled marine carbon. Many of these microorganisms have the genetic potential to fix inorganic carbon (autotrophs) or assimilate single-carbon compounds (methylotrophs). We identified the functions of autotrophic and methylotrophic microorganisms in a vent plume at Axial Seamount, where hydrothermal activity provides a biogeochemical hot spot for carbon fixation in the dark ocean. Free-living members of the SUP05/Arctic96BD-19 clade of marine gamma-proteobacterial sulfur oxidizers (GSOs) are distributed throughout the northeastern Pacific Ocean and dominated hydrothermal plume waters at Axial Seamount. Marine GSOs expressed proteins for sulfur oxidation (adenosine phosphosulfate reductase, sox (sulfur oxidizing system), dissimilatory sulfite reductase and ATP sulfurylase), carbon fixation (ribulose-1,5-bisphosphate carboxylase oxygenase (RuBisCO)), aerobic respiration (cytochrome c oxidase) and nitrogen regulation (PII). Methylotrophs and iron oxidizers were also active in plume waters and expressed key proteins for methane oxidation and inorganic carbon fixation (particulate methane monooxygenase/methanol dehydrogenase and RuBisCO, respectively). Proteomic data suggest that free-living sulfur oxidizers and methylotrophs are among the dominant primary producers in vent plume waters in the northeastern Pacific Ocean.     

Cytoplasmic sulfurtransferases in the purple sulfur bacterium Allochromatium vinosum: evidence for sulfur transfer from DsrEFH to DsrC

While the importance of sulfur transfer reactions is well established for a number of biosynthetic pathways, evidence has only started to emerge that sulfurtransferases may also be major players in sulfur-based microbial energy metabolism. Among the first organisms studied in this regard is the phototrophic purple sulfur bacterium Allochromatium vinosum. During the oxidation of reduced sulfur species to sulfate this Gammaproteobacterium accumulates sulfur globules. Low molecular weight organic persulfides have been proposed as carrier molecules transferring sulfur from the periplasmic sulfur globules into the cytoplasm where it is further oxidized via the "Dsr" (dissimilatory sulfite reductase) proteins. We have suggested earlier that the heterohexameric protein DsrEFH is the direct or indirect acceptor for persulfidic sulfur imported into the cytoplasm. This proposal originated from the structural similarity of DsrEFH with the established sulfurtransferase TusBCD from E. coli. As part of a system for tRNA modification TusBCD transfers sulfur to TusE, a homolog of another crucial component of the A. vinosum Dsr system, namely DsrC. Here we show that neither DsrEFH nor DsrC have the ability to mobilize sulfane sulfur directly from low molecular weight thiols like thiosulfate or glutathione persulfide. However, we demonstrate that DsrEFH binds sulfur specifically to the conserved cysteine residue DsrE-Cys78 in vitro. Sulfur atoms bound to cysteines in DsrH and DsrF were not detected. DsrC was exclusively persulfurated at DsrC-Cys111 in the penultimate position of the protein. Most importantly, we show that persulfurated DsrEFH indeed serves as an effective sulfur donor for DsrC in vitro. The active site cysteines Cys78 of DsrE and Cys20 of DsrH furthermore proved to be essential for sulfur oxidation in vivo supporting the notion that DsrEFH and DsrC are part of a sulfur relay system that transfers sulfur from a persulfurated carrier molecule to the dissimilatory sulfite reductase DsrAB.     

Hydrogenogenic and sulfidogenic growth of <Emphasis Type="Italic">Thermococcus</Emphasis> archaea on carbon monoxide and formate

Enrichment and pure cultures of hyperthermophilic archaea capable of anaerobic growth on one-carbon compounds (CO and/or formate) were obtained from deep-sea sites of hydrothermal activity at the Mid-Atlantic Ridge, Lau Basin, and Guaymas Basin. All isolates belonged to the T. barophilus?T. paralvinellae group within the genus Thermococcus. In all cases available for analysis, the genomes of Thermococcus strains capable of growth by hydrogenogenic utilization of CO and/or formate contained clusters of genes encoding energy-converting hydrogenase and either CO dehydrogenase or formate dehydrogenase and formate transporter. Apart from the previously known processes of hydrogenogenic oxidation of CO and formate, the oxidation of these substrates coupled to sulfur reduction was observed, processes previously unknown among archaea. The capacities for hydrogenogenic or sulfidogenic oxidation of CO and formate occurred in the studied strains in all possible combinations, which could only in part be explained by peculiarities of organization of genetic determinants revealed in the genomes. Investigation of CO and formate consumption kinetics revealed that T. barophilus strain Ch5 was able to grow at concentrations close to the environmental ones. Thus, it was shown that hyperthermophilic archaea from deep-sea hydrothermal vents are able to utilize one-carbon substrates of abiotic origin both in the presence of an electron acceptor (sulfur) and in its absence. These processes were probably of importance under the conditions of the early Earth biosphere.     

Sulfur oxidation activities of pure and mixed thermophiles and sulfur speciation in bioleaching of chalcopyrite

The sulfur oxidation activities of four pure thermophilic archaea Acidianus brierleyi (JCM 8954), Metallosphaera sedula (YN 23), Acidianus manzaensis (YN 25) and Sulfolobus metallicus (YN 24) and their mixture in bioleaching chalcopyrite were compared. Meanwhile, the relevant surface sulfur speciation of chalcopyrite leached with the mixed thermophilic archaea was investigated. The results showed that the mixed culture, with contributing significantly to the raising of leaching rate and accelerating the formation of leaching products, may have a higher sulfur oxidation activity than the pure cultures, and jarosite was the main passivation component hindering the dissolution of chalcopyrite, while elemental sulfur seemed to have no influence on the dissolution of chalcopyrite. In addition, the present results supported the former speculation, i.e., covellite might be converted from chalcocite during the leaching experiments, and the elemental sulfur may partially be the derivation of covellite and chalcocite.     

Microbial Nitrogen and Sulfur Cycles at the Gypsum Dunes of White Sands National Monument,New Mexico

The White Sands National Monument from New Mexico (U.S.A) contains one of the largest known gypsum dune fields with unique, rapidly migrating, arid, evaporitic habitats. Deposits from dune sides and interdune areas were collected in order to determine the characteristics of microbial habitat and communities through mineral assemblages, microbial pigments along with investigations of nitrogen and sulfur cycles. The most abundant pigments, scytonemin and carotenoids, were common UV protective pigments. Predominance of nitrite and nitrate over ammonium nitrogen (2.16: 1) implies that nitrification processes might be important in this ecosystem. Ammonium oxidizers from groups of β-, γ-proteobacteria and archaea were detected in all deposits, thereby indicating microbial involvement in nitrification. Additionally, denitrifying organisms with nirS and nirK genes were also present in most of the analyzed samples. The presence of trace carbonate mineral phases in association with biofilm implies possible microbial sulfate reduction. Microbes with metabolic abilities for sulfur cycling (i.e., dissimilatory sulfite reducers, purple sulfur bacteria, green sulfur and non-sulfur bacteria, and organisms with the APS enzyme) were identified in all samples. These particular organisms have the ability to reduce sulfate and to re-oxidize reduced sulfur compounds back to sulfate.