嗜酸硫杆菌在工农业中的应用

嗜酸硫杆菌(属)(Acidithiobacillus spp.)能够氧化亚铁、硫或还原性无机硫化合物(reduced inorganic sulfur compounds,RISCs)获得能量,固定二氧化碳,是一类典型的嗜酸性化能自养微生物。嗜酸硫杆菌广泛分布于酸性矿水、热泉等酸性环境中,是地球生态系统硫和铁元素循环的主要推动者。嗜酸硫杆菌独特的生理代谢特征和极端环境适应性,使其广泛应用于生物浸出领域。本文综述了嗜酸硫杆菌的生理代谢特征和极端环境下的适应机制,阐述了嗜酸硫杆菌在工农业中的应用,讨论了面向国家重大需求,嗜酸硫杆菌在今后的主要研究方向和需要解决的关键科学问题,为嗜酸硫杆菌在生理代谢、环境适应和工农业应用的研究提供重要的线索和启示。     

隐藏嗜酸菌Acidiphilium cryptum的研究进展

分离自酸性矿坑水的隐藏嗜酸菌Acidiphilium cryptum具有化能兼养型细菌的典型特征,在缺乏有机质的环境中能启用体内硫氧化代谢途径营化能自养,进行硫化矿物的分解;而在有机质存在时该菌营异养生长,并在体内合成聚β羟基丁酸酯(polyhydroxybutyrate,PHB),此外在酸性条件下对Fe(III)和Cr(VI)等金属离子具有还原作用,并具有发展微生物燃料电池的潜力.该菌因其独特的生理特性及在浸矿微生物群落中的显著生态地位而在生物冶金、环境修复、生物材料等领域具有重要的经济和社会意义.对隐藏嗜酸菌的生物学特性、分离培养方法、重要特性的研究进展以及应用等方面的国内外研究现状进行综述,并对其应用前景和研究趋势进行展望.     

硫杆菌的分子遗传学研究

硫杆菌（丁litKucjllSS）是一类革兰氏阴性的化能无机营养细菌,也是分布最广,研究得最多,经济意义很大的硫氧化细菌。该属中的氧化硫硫杆菌（T．th。whns）和氧化亚铁流杆菌（T．fer－rtx）xlua叫是极端嗜酸性的专性自养细菌,最适因为2,0～3．5,广泛分布于流化矿床的酸性废水中。这类细菌很早就被广泛应用于贵重金属的浸出和回收,特别适合于从低品位的矿石中浸出稀有资重金属,主要是通过氧化FeSO和Fe＆产生Fe”氧化剂,氧化金属硫化物使之变成可溶性的硫酸盐形式,从而回收有用金属。这种方法具有低成本,低能耗,无污染等特点…     

嗜酸性硫杆菌群体感应系统研究进展

嗜酸性硫杆菌(Acidithiobacillus spp.)是一类重要的极端环境微生物与工业微生物。该类细菌通过氧化硫或亚铁获得电子以固定二氧化碳进行自养生长,是驱动矿山环境酸化和重金属溶出的关键菌群,也是生物冶金等微生物浸出技术中的核心菌群。群体感应(quorum sensing, QS)系统是细菌种内及种间信息交流的重要方式,广泛分布于嗜酸性硫杆菌等化能自养微生物中,比如类似于LuxI/R的AfeI/R系统。系统介绍近年来嗜酸性硫杆菌菌体感应系统研究成果,尤其是在AfeI/R种群分布、生物学功能、调节机制及其应用研究中的新发现与新理论。讨论今后嗜酸性硫杆菌群体感应系统研究的主要方向及需要解决的关键科学问题,以促进极端微生物群体感应系统理论研究的开展与产业应用技术的开发。     

硫代硫酸盐氧化菌TX的分离、鉴定及其生物学特性

从聚硫橡胶废水处理系统中分离到一株硫代硫酸盐氧化细菌TX.根据其形态学特征、生理特征和16S rRNA基因序列相似性分析,将该菌株初步鉴定为盐生硫杆菌属(Halothiobacillus sp.)(GenBank登录号为EU871645).该菌株能利用硫代硫酸盐、单质硫、连四硫酸盐、硫化物或亚硫酸盐为唯一能源进行自养生长,不能利用葡萄糖、蔗糖、果糖、乳糖、麦芽糖或酵母粉进行异养生长,为专性化能无机自养型硫杆菌.在以硫代硫酸钠为唯一能源的培养基中其最适生长温度为30℃～35℃,最适起始pH值为3.0～5.0.在矿物盐培养基中,硫代硫酸盐最终被氧化成硫酸,造成培养基pH持续下降.在摇瓶分批培养和硫胶废水处理过程中均检测到连四硫酸盐的积累,表明该菌株主要通过连四硫酸盐途径或"S4I"途径进行硫代硫酸盐的生物氧化.     

浸矿细菌的遗传工程

硫杆菌属(Thiobacillus)的一些种,氧化硫硫杆菌(T.thiooxidans)和氧化亚铁硫杆菌(T.ferrooxidans),广泛分布于硫化矿床的酸性矿水中,最适生长pH为2.0—2.5,是一类革兰氏阴性专性自养细菌。这类细菌广泛应用于有用金属的浸出,特别适合于从低品位的     

抗砷载体的构建及在喜温硫杆菌中的接合转移

利用DNA体外重组技术,将质粒载体pUM3上的抗砷基因片段亚克隆到含有强启动子(tae启动子)并具有广泛寄主范围特性的IncQ族质粒pMMB24上,成功构建了含有强启动子的抗砷质粒pSDRA3,以及删除调节基因片段的组成型表达的抗砷质粒pSDRA4。通过接合转移的方式将其导入专性自养极端嗜酸性的喜温硫杆菌中,首次成功地建立了喜温硫杆菌的遗传转移系统,构建了冶金工程菌T.caldus(pSDRA3)和T．caldus(pSDRA4)。与野生菌相比,重组菌抗砷性能明显提高。     

环境对沼泽红假单胞菌基因型的影响

<正>紫色光合细菌是不产氧光合细菌的一个重要分支,包括紫色非硫细菌和紫色硫细菌。其中紫色非硫细菌具有极其丰富的代谢模式,可进行光能异养、光能自养和化能异养生长,其代谢的多样性使得它们广泛存活于不同的生态系统中,如土壤、湖泊、海洋及底泥等[1-2]。沼泽红假单胞菌(Rhodopseudomonas palustris)是紫     

一株中度嗜热嗜酸铁氧化细菌特性研究*

从我国煤矿废石堆分离到一株中度嗜热嗜酸铁氧化细菌MLY菌株,最适生长温度50℃－54℃,最适pH1.2-1.4。MLY菌株是兼性化能自养菌,能利用酵母粉异养生长。在自养和混合营养条件下,能氧化Fe^2 、黄铁矿(FeS2)和元素硫(S^0)。自养营养时,氧化元素硫较弱。对比研究MLY菌株和氧化亚铁硫杆菌(Thiobacillus ferrooxidans)A10菌株对Fe^2 和黄铁矿的氧化作用,结果表明,MLY比A10的氧化速度快1倍多。     

嗜酸硫氧化细菌消解元素硫的研究进展

浸矿酸性环境下,金属硫化矿在Fe3+作用下,经过硫代硫酸盐途径或多聚硫化氢途径而分解的过程中导致大量元素硫的累积,进而可能在金属硫化矿表面形成疏水元素硫层,阻碍金属离子的进一步浸出。酸性环境下,惰性元素硫的消解必须借助嗜酸硫氧化细菌来实现。该消解过程包括嗜酸硫氧化细菌对元素硫的吸附、转运以及氧化转化等过程。本文对近年来嗜酸硫氧化细菌消解元素硫过程的相关研究进行了全面评述,认为有关嗜酸硫氧化细菌消解元素硫的分子机制的清晰阐述还有待人们通过对消解过程的各个环节的分子机制进行大量研究来实现。     

Draft genome sequence of the extremely acidophilic biomining bacterium Acidithiobacillus thiooxidans ATCC 19377 provides insights into the evolution of the Acidithiobacillus genus

Acidithiobacillus thiooxidans is a mesophilic, extremely acidophilic, chemolithoautotrophic gammaproteobacterium that derives energy from the oxidation of sulfur and inorganic sulfur compounds. Here we present the draft genome sequence of A. thiooxidans ATCC 19377, which has allowed the identification of genes for survival and colonization of extremely acidic environments.     

Acidocella aromatica sp. nov.: an acidophilic heterotrophic alphaproteobacterium with unusual phenotypic traits

Three obligately heterotrophic bacterial isolates were identified as strains of a proposed novel species of extremely acidophilic, mesophilic Alphaproteobacteria, Acidocella aromatica. They utilized a restricted range of organic substrates, which included fructose (but none of the other monosaccharides tested), acetate and several aromatic compounds (benzoate, benzyl alcohol and phenol). No growth was obtained on complex organic substrates, such as yeast extract and tryptone. Tolerance of the proposed type strain of the species (PFBC) to acetic acid was much greater than that typically reported for acidophiles. The bacteria grew aerobically, and catalyzed the dissimilatory reductive dissolution of the ferric iron mineral schwertmannite under both micro-aerobic and anaerobic conditions. Strain PFBC did not grow anaerobically via ferric iron respiration, though it has been reported to grow in co-culture with acid-tolerant sulfidogenic bacteria under strictly anoxic conditions. Tolerance of strains of Acidocella aromatica to nickel were about two orders of magnitude greater than those of other Acidocella spp., though similar levels of tolerance to other metals tested was observed. The use of this novel acidophile in solid media designed to promote the isolation and growth of other (aerobic and anaerobic) acidophilic heterotrophs is discussed.     

Adaptive responses of chemolithoautotrophic acidophilic Acidithiobacillus ferrooxidans to sewage sludge

AIM: The aim of the present study was to investigate the phenotypic and genotypic variability of two strains of Acidithiobacillus ferrooxidans genus during growth in sewage sludge. METHODS AND RESULTS: Compared with A. ferrooxidans cells grown in mineral medium, those grown in sewage sludge demonstrated remarkable changes in ultrastructure (transmission electron microscopy) and significantly elongated lag phases. These latter cells also lacked carboxysomes and rusticyanin, showed lower level of cytochromes and exhibited modifications to their outer membrane proteins (SDS-PAGE). Restriction fragment length polymorphism analysis using pulsed-field gel electrophoresis showed that most restriction fragments were highly conserved and shared by strains grown under different conditions. However, in relation to cells grown in mineral medium, sludge-grown A. ferrooxidans lacked a number of restriction fragments, clearly indicating structural changes to the chromosomal DNA of the organism. CONCLUSIONS: In combination, the results of this study provide evidence of adaptive responses by chemolithoautotrophic acidophilic A. ferrooxidans to facilitate growth in sewage sludge. SIGNIFICANCE AND IMPACT OF THE STUDY: The obtained results are important from scientific as well as industrial application point of view, because they confirmed that A. ferrooxidans highly sensitive to organic compounds bacteria is useful in biotechnologies of heavy metal removal from shale ore, polluted soils and sewage sludge containing organic hazardous compounds.     

Enzymes of Carbohydrate Metabolism in Thiobacillus species

A study was made of enzymes of carbohydrate metabolism in representative thiobacilli grown with and without glucose. The data show that Thiobacillus perometabolis possesses an inducible Entner-Doudoroff pathway and is thus similar to T. intermedius and T. ferrooxidans. T. novellus lacks this pathway. Instead, a non-cyclic pentose phosphate pathway along with the Krebs cycle is apparently the major route of glucose dissimilation in this organism. Glucose does not support or stimulate the growth of strains of T. neapolitanus, T. thioparus, and T. thiooxidans examined, nor does its presence in the growth medium greatly influence their enzymatic constitution. These obligately chemolithotrophic thiobacilli do not possess the Entner-Doudoroff pathway. Their nicotinamide adenine dinucleotide (NAD)-linked isocitrate dehydrogenase activity predominates over their nicotinamide adenine dinucleotide phosphate (NADP)-linked activity; the converse is true for the other thiobacilli. The data suggest that NAD-linked isocitrate dehydrogenase activity in thiobacilli is involved in biosynthetic reactions.     

Methylated sulfur compounds in microbial mats: in situ concentrations and metabolism by a colorless sulfur bacterium

The concentrations of the volatile organic sulfur compounds methanethiol, dimethyl disulfide, and dimethyl sulfide (DMS) and the viable population capable of DMS utilization in laminated microbial ecosystems were evaluated. Significant levels of DMS and dimethyl disulfide (maximum concentrations of 220 and 24 nmol cm3 of sediment-1, respectively) could be detected only at the top 20 mm of the microbial mat, whereas methanethiol was found only at depth horizons from 20 to 50 mm (maximum concentration of 42 nmol cm3 of sediment-1). DMS concentrations in the surface layer doubled after cold hydrolysis of its precursor, dimethylsulfoniopropionate. Most-probable-number counts revealed 2.2 x 10(5) cells cm3 of sediment-1, in the 0- to 5-mm depth horizon, capable of growth on DMS as the sole source of energy. An obligately chemolithoautotrophic bacillus designated strain T5 was isolated from the top layer of the marine sediment. Continuous culture studies in which DMS was the growth-limiting substrate revealed a maximum specific growth rate of 0.10 h-1 and a saturation constant of 90 mumol liter-1 for aerobic growth on this substrate.     

Methylated sulfur compounds in microbial mats: in situ concentrations and metabolism by a colorless sulfur bacterium.

The concentrations of the volatile organic sulfur compounds methanethiol, dimethyl disulfide, and dimethyl sulfide (DMS) and the viable population capable of DMS utilization in laminated microbial ecosystems were evaluated. Significant levels of DMS and dimethyl disulfide (maximum concentrations of 220 and 24 nmol cm3 of sediment-1, respectively) could be detected only at the top 20 mm of the microbial mat, whereas methanethiol was found only at depth horizons from 20 to 50 mm (maximum concentration of 42 nmol cm3 of sediment-1). DMS concentrations in the surface layer doubled after cold hydrolysis of its precursor, dimethylsulfoniopropionate. Most-probable-number counts revealed 2.2 x 10(5) cells cm3 of sediment-1, in the 0- to 5-mm depth horizon, capable of growth on DMS as the sole source of energy. An obligately chemolithoautotrophic bacillus designated strain T5 was isolated from the top layer of the marine sediment. Continuous culture studies in which DMS was the growth-limiting substrate revealed a maximum specific growth rate of 0.10 h-1 and a saturation constant of 90 mumol liter-1 for aerobic growth on this substrate.     

Characterization of a new metal-mobilizing Thiobacillus isolate

A new acidophilic, mineral sulphide oreoxidizing bacterium was isolated from a uranium mine near Salamanca, Spain. Cells were rod-shaped, motile and gram-negative. They were aerobes, could grow on pyrite and use sulphur or thiosulphate as sole energy source, suggesting this new isolate belongs to the genus Thiobacillus. It could grow neither with glucose nor with yeast extract as sole substrates. It could not grow on ferrous sulphate as the only energy source, although it grew in the same medium supplemented with glucose, yeast extract or thiosulphate. It was a mesophilic and extremely acidophilic Thiobacillus, with an optimal pH of 1.5 2. The G+C content of the DNA was 58%. The new isolate could grow in cultures on pyrite where electrophoretic pattern was clearly different from those of other thiobacilli, such as T. ferrooxidans.Abbreviations G+C Guanine + Cytosine     

Isolation of an obligately chemolithoautotrophic,halophilic and aerobic hydrogen-oxidizing bacterium from marine environment

An obligately chemolithoautotrophic and aerobic hydrogen-oxidizing bacterium was isolated from seawater of the Shonan Coast, Kanagawa Pref., Japan. The isolate was a Gram-negative, comma-shaped rod cell measuring 0.2 to 0.5 by 1 to 2 m. The cells occurred singly and were motile by a polar flagellum. The deoxyribonucleic acid base composition of the isolate was 44.1 mol% guanine plus cytosine. The optimal temperature for autotrophic growth on H₂–O₂–CO₂ was around 37°C, and no growth was observed at 5° C or 45° C. The optimal pH for growth was around 6.5. NaCl was required for growth with an optimum of 0.5 M. Elemental sulfur, thiosulfate or tetrathionate was utilized, as well as molecular hydrogen, as the sole energy source. No heterotrophic growth was observed on organic media tested. To our knowledge, this is the first report of the isolation of a marine, aerobic hydrogen-oxidizing bacterium, and of an obligately chemolithoautotrophic, mesophilic and aerobic hydrogen-oxidizing bacterium.     

Growth of Ferrobacillus ferrooxidans on Organic Matter

Following a brief adaptation period to glucose, Ferrobacillus ferrooxidans was grown on glucose, mannitol, several other sugars, and a few amino acids in the absence of an oxidizable iron source. Prolonged growth on an organic substrate free from iron rendered the organism obligately organotrophic. The growth rate of the bacterium was greater in heterotrophic culture; the doubling time was approximately 4.5 hr on glucose. The bacterium retained its acidophilic properties during adaptation and growth on glucose and would not grow in neutral or slightly alkaline media. Addition of p-aminobenzoic acid was necessary for abundant growth of the cells on glucose. Of the eight strains of Fe(++)-oxidizing bacteria studied, only two strains grew on glucose in a nondialyzed system. The results of manometric studies are discussed with regard to metabolic efficiency of organic matter in this organism.     

The Genome of Deep-Sea Vent Chemolithoautotroph Thiomicrospira crunogena XCL-2

Presented here is the complete genome sequence of Thiomicrospira crunogena XCL-2, representative of ubiquitous chemolithoautotrophic sulfur-oxidizing bacteria isolated from deep-sea hydrothermal vents. This gammaproteobacterium has a single chromosome (2,427,734 base pairs), and its genome illustrates many of the adaptations that have enabled it to thrive at vents globally. It has 14 methyl-accepting chemotaxis protein genes, including four that may assist in positioning it in the redoxcline. A relative abundance of coding sequences (CDSs) encoding regulatory proteins likely control the expression of genes encoding carboxysomes, multiple dissolved inorganic nitrogen and phosphate transporters, as well as a phosphonate operon, which provide this species with a variety of options for acquiring these substrates from the environment. Thiom. crunogena XCL-2 is unusual among obligate sulfur-oxidizing bacteria in relying on the Sox system for the oxidation of reduced sulfur compounds. The genome has characteristics consistent with an obligately chemolithoautotrophic lifestyle, including few transporters predicted to have organic allocrits, and Calvin-Benson-Bassham cycle CDSs scattered throughout the genome.