海洋滩涂沉积物环境中几类主要细菌的动态分布

目的研究宁波市宁海县贝类养殖滩涂中异养细菌、反硝化细菌、氨化细菌和硫酸还原菌等几类细菌的动态分布及异养细菌的菌群组成。方法异养细菌的计数采用平板菌落计数法;反硝化细菌、氨化细菌和硫酸还原菌的计数采用MPN法;并参照Oliver提供的海洋细菌鉴定检索图、《海洋调查规范》和《一般细菌常用鉴定方法》提供的图式将分离到的细菌鉴定至属。结果滩涂沉积物环境中异养细菌的数量波动在8.00×103~7.60×104CFU/g(湿重),反硝化细菌的数量变动在5.00×102~5.00×104个/g(湿重),氨化细菌的数量变动在3.00×105~5.00×107个/g(湿重),硫酸还原菌的数量分布在9.00×104~9.00×106个/g(湿重),温度的变化对这几类细菌的数量分布没有直接的影响。所分离的细菌经过鉴定,可归于15个属与肠杆菌科的部分属,其中肠杆菌科(Enterobacteriaceae)的部分属、梭状芽胞杆菌属(Clostridium)、芽胞杆菌属(Bacillus)、棒状杆菌属(Corynebacterium)、发光杆菌属(Photobacterium)、假单胞菌属(Pseudomonas)等为优势菌属。沉积物中氨化细菌、反硝化细菌和硫酸还原菌的检出率均为100%,且含量很高。结论经过多年不间歇的养殖后,宁海贝类养殖滩涂已经不同程度地受到了污染,应尽快调整养殖容量并对养殖环境进行重新布局。     

Removal of sulphate, COD and Cr(VI) in simulated and real wastewater by sulphate reducing bacteria enrichment in small bioreactor and FTIR study

The present study was conducted to investigate the chromium(VI), COD and sulphate removal efficiency from aqueous solution and treatment of real effluent (CETP) in a small scale bioreactor using sulphate reducing bacteria consortium. Effect of different hydraulic retention times (HRTs), initial metal concentrations, various carbon sources and temperatures were studied on removal of chromium(VI), COD and sulphate. Maximum chromium(VI) and sulphate removal was found to be 96.0% and 82.0%, respectively, at initial concentration of 50 mg l⁻¹ using lactate as carbon source. However, highest COD removal was 36.2% in medium containing fructose as the carbon source and electron donor. NADH dependent chromate reductase activity was not observed which indicated the anaerobic consortium. Initially consortium medium with a strong negative oxidation reduction potential indicated the reducing activity. The FTIR spectrum of the sulphate reducing bacteria consortium clearly shows the existence of the sulphate ions and signifies that sulfate reducing bacteria have used sulfate during the growth phase.     

缢蛏滩涂养殖环境的细菌群落组成及分析

1引言滩涂贝类养殖由于实行生态养殖,目前已成为宁波市海水养殖业的一大热点.近几年来,随着沿海经济的高速发展,滩涂环境污染问题日趋严重,养殖贝类大规模死亡的现象时有发生,给海洋水产经济造成了严重损失.而微生物数量及种群分布与区域环境有着密切联系,它可直接或间接地反映区域环境的营养水平、污染程度及底质状况等.目前有关养殖环境微生物学的研究主要是针对水体及养殖池底泥中的细菌[1,4,7～10],而对养殖滩涂环境细菌的研究很少[15,18,19].为此,本文以宁波市昆亭缢蛏滩涂养殖区为研究对象,从细菌生态学角度对滩涂缢蛏养殖环境进行研…     

Toxic effects exerted on methanogenic, nitrifying and denitrifying bacteria by chemicals used in a milk analysis laboratory

The toxic effects caused by the chemicals contained in wastewaters generated by laboratories involved in raw milk analyses were assessed using batch assays. These assays were carried out separately with methanogenic, ammonium-oxidizing, nitrite-oxidizing and denitrifying bacteria. Since sodium azide is one of the main components of the chemical mixture present in these streams, a set of assays was carried out with the whole chemical mixture, and another one was performed only with azide as the sole toxicant. The concentrations of all chemicals in the raw wastewaters (100%) were the fundamental references used to assess the relative concentrations corresponding to a decrease of 50% in bacterial activity (IC₅₀). The results obtained showed that nitrite-oxidizing bacteria were the most sensitive microorganisms, with IC₅₀ relative concentrations around 0.04%. The values obtained for the other groups were: 20, 20 and 33% for methanogenic, ammonium-oxidizing and denitrifying bacteria, respectively.     

The distribution and activity of sulphate reducing bacteria in estuarine and coastal marine sediments

Sulphate-reducing bacteria (SRB) play a vital role both the carbon and sulphur cycles and thus are extremely important components of the global microbial community. However, it is clear that the ecology, the distribution and activity of different SRB groups is poorly understood. Probing of rRNA suggests that different sediments have distinctly different patterns of SRB with complex factors controlling the activity of these organisms. The linking of community structure and function using sediment slurry microcosms suggests that certain groups of SRB, e.g., Desulfobacter and Desulfobulbus, can be linked to the use of specific substrates in situ. However, it is still unclear what environmental substrates are utilised by the majority of known SRBs. The work to date has greatly enhanced our understanding of the ecology of these organisms and is beginning to suggest patterns in their distribution and activity that may be relevant to understanding microbial ecology in general. This revised version was published online in August 2006 with corrections to the Cover Date.     

Methane production from organic acid-rich wastewaters by immobilized thermophilic methane-producing bacteria

Thermophilic methane-producing bacteria isolated from a wastewater treatment facility have been immobilized in acetylcellulose filter with agar. The immobilized cells produced methane from wastewaters in rich organic acid (acetic, propionic and butyric acids) at the rate of 1.4 μmol mg protein⁻¹ h⁻¹. The optimum conditions for methane production by immobilized whole cells were 52–55°C and pH 7.0–8.0. The immobilized cells retained 80% of the initial activity after exposure to air. The immobilized thermophilic bacteria produced methane continuously over 10 days at 52°C.     

Sulfate reduction from phosphogypsum using a mixed culture of sulfate-reducing bacteria

Phosphogypsum (CaSO₄), a primary by-product of phosphoric acid production, is accumulated in large stockpiles and occupies vast areas of land. It poses a severe threat to the quality of water and land in countries producing phosphoric acid. In this study, the potential of sulfate-reducing bacteria for biodegradation of this sulfur-rich industrial solid waste was assessed. The effect of phosphogypsum concentration, carbon and nitrogen sources, temperature, pH and stirring on the growth of sulfate-reducing bacteria was investigated. Growth of sulfate-reducing bacteria was monitored by measuring sulfide production. Phosphogypsum was shown to be a good source of sulfate, albeit that the addition of organic carbon was necessary for bacterial growth. Biogenic sulfide production occurred with phosphogypsum up to a concentration of 40 g L⁻¹, above which no growth of sulfate-reducing bacteria was observed. Optimal growth was obtained at 10 g L⁻¹ phosphogypsum. Both the gas mixture H₂/CO₂ and lactate supported high amounts of H₂S formation (19 and 11 mM, respectively). The best source of nitrogen for sulfate-reducing bacteria was yeast extract, followed by ammonium chloride. The presence of nitrate had an inhibitory effect on the process of sulfate reduction. Stirring the culture at 150 rpm slightly stimulated H₂S formation, probably by improving sulfate solubility.     

Sulphate respiration from hydrogen in Desulfovibrio bacteria: a structural biology overview

Sulphate-reducing organisms are widespread in anaerobic enviroments, including the gastrointestinal tract of man and other animals. The study of these bacteria has attracted much attention over the years, due also to the fact that they can have important implications in industry (in biocorrosion and souring of oil and gas deposits), health (in inflamatory bowel diseases) and the environment (bioremediation). The characterization of the various components of the electron transport chain associated with the hydrogen metabolism in Desulfovibrio has generated a large and comprehensive list of studies. This review summarizes the more relevant aspects of the current information available on the structural data of various molecules associated with hydrogen metabolism, namely hydrogenases and cytochromes. The transmembrane redox complexes known to date are also described and discussed. Redox-Bohr and cooperativity effects, observed in a few cytochromes, and believed to be important for their functional role, are discussed. Kinetic studies performed with these redox proteins, showing clues to their functional inter-relationship, are also addressed. These provide the groundwork for the application of a variety of molecular modelling approaches to understanding electron transfer and protein interactions among redox partners, leading to the characterization of several transient periplasmic complexes. In contrast to the detailed understanding of the periplasmic hydrogen oxidation process, very little is known about the cytoplasmic side of the respiratory electron transfer chain, in terms of molecular components (with exception of the terminal reductases), their structure and the protein–protein interactions involved in sulphate reduction. Therefore, a thorough understanding of the sulphate respiratory chain in Desulfovibrio remains a challenging task.     

Inhibition of methanogenesis by sulphate reducing bacteria competing for transferred hydrogen

A methanogenic bacterial consortium was obtained after inoculation of benzoate medium under N₂/CO₂ atmosphere with intertidal sediment. A hydrogen donating organotroph andMethanococcus mazei were isolated from this enrichment. H₂-utilising sulphate reducing bacteria were isolated under H₂/CO₂ in the absence of organic electron donors. TheMethanococcus was able to produce methane in yeast extract medium under N₂/CO₂ if the H₂ donating organism was present, and sulphate reduction occurred if the hydrogen utilising sulphate reducing bacteria were grown with the H₂ donating organism. The ability of the H₂ utilising sulphate reducing bacteria to inhibitMethanococcus competitively was shown in cultures containing both of these H₂ utilising bacteria.Abbreviations HDO hydrogen donating organism - SRB sulphate reducing bacteria - HSRB hydrogen utilising sulphate reducing bacteria     

两种光合细菌生物转化槲寄生培养液菌体中几种同工酶的变化

采用聚丙烯酰胺凝胶电泳法对两种光合细菌的生物转化槲寄生培养液中菌体的蛋白质和几种同工酶进行研究,并以纯光合细菌培养液中菌体作对照。结果表明,光合细菌生物转化槲寄生过程中,两种光合细菌的蛋白质、酯酶同工酶和过氧化物酶同工酶均发生改变,某些蛋白质、酯酶和过氧化物酶的合成受到抑制,并有新的蛋白质、酯酶和过氧化物酶生成;超氧化物歧化酶的表达未明显改变。由此可见,槲寄生能诱导光合细菌合成新的酯酶和过氧化物酶,这些诱导酶可能参与了槲寄生的生物转化。为光合细菌生物转化槲寄生转化机理的研究及槲寄生在抗肿瘤领域的进一步应用奠定了基础。     

人肠道菌对天然产物代谢及转化的研究进展

天然产物的人肠道菌代谢及转化研究已引起各国学者的重视,并作为热点课题进行了大量研究工作。本文对近二十年来各国学者关于黄酮类、萜类、苯丙素类、生物碱类、甾体类及其他天然产物的人肠道菌代谢及转化研究工作进行了综述,总结了各类天然产物在人肠道菌作用下的代谢路径及转化规律,以期为该领域的进一步深入研究提供参考。     

Chemical Purity of Shewanella oneidensis-Induced Magnetites

Magnetite is a common iron oxide produced both inorganically and biogenically. Biologically-induced magnetite is often originated, under appropriate conditions, as a result of the Fe³⁺ reduction by dissimilatory iron reducing bacteria, which are usually found in anoxic environments or at the oxic-anoxic interface. Such a Fe³⁺ bioreduction occurs upon this cation acting as an electron acceptor of an anaerobic respiration, thus creating favorable conditions for magnetite precipitation. This biologically-induced magnetite is an important biomineral in the environments inhabited by iron reducing bacteria. The presence of a variety of cations may influence both the biomineralization process and the resulting biomineral, however this phenomenon has not been investigated extensively. In the present study, we study the effect on the magnetite biomineralization process of the presence of calcium, magnesium and manganese in the culture medium where Shewanella oneidensis lives. We also test the incorporation of these cations into the crystalline structure of inorganic and biogenic magnetite induced by S. oneidensis. According to our findings, manganese ions likely become incorporated into the crystal structure of biologically produced magnetites, while magnesium ions are incorporated in inorganic magnetites, and calcium ions are excluded from the crystal structure of both inorganic and biotic magnetites. We hypothesize that the incorporation of cations into magnetite depends not only on the relative cation radii, but also on the mechanisms of magnetite formation.     

Vinyl ketone reduction by three distinct Gluconobacter oxydans 621H enzymes

Three cytosolic NADPH-dependent flavin-associated proteins (Gox2107, Gox0502, and Gox2684) from Gluconobacter oxydans 621H were overproduced in Escherichia coli, and the recombinant enzymes were purified and characterized. Apparent native molecular masses of 65.2, 78.2, and 78.4 kDa were observed for Gox2107, Gox0502, and Gox2684, corresponding to a trimeric structure for Gox2107 and dimers for Gox0502 and Gox2684. Analysis of flavin content revealed Gox2107 was flavin adenine dinucleotide dependent, whereas Gox0502 and Gox2684 contained flavin mononucleotide. The enzymes were able to reduce vinyl ketones and quinones, reducing the olefinic bond of vinyl ketones as shown by ¹H nuclear magnetic resonance. Additionally, Gox0502 and Gox2684 stereospecifically reduced 5S-(+)-carvone to 2R,5S-dihydrocarvone. All enzymes displayed highest activities with 3-butene-2-one and 1,4-naphthoquinone. Gox0502 and Gox2684 displayed a broader substrate spectrum also reducing short-chain α-diketones, whereas Gox2107 was most catalytically efficient.     

用硫酸盐还原菌处理重金属废水的研究

介绍了用硫酸盐还原菌处理重金属废水的几种主要方法和原理。硫酸盐还原菌处理含重金属废水主要是通过将可溶性的重金属离子转化成不溶性的金属硫化物、氢氧化物、碳酸盐的方式 ,或直接通过以菌体对重金属离子的吸附完成的。目前研究用硫酸盐还原菌处理重金属废水的主要方法有分批沉淀工艺、吸附处理工艺、化学法和硫酸盐还原菌的混合工艺、全混合处理工艺及硫酸盐还原菌的厌氧上流式污泥床和流化床工艺 ,并对其主要的工艺指标进行了比较。     

Kinetics of SO4−2 reduction under different growth media by sulfate reducing bacteria

Sulfate Reducing Bacteria (SRB) were used to reduce the SO₄^–2 concentration in waste water. The growth pattern of SRB was found by varying the concentration of nutrients and the biomass. The specific reaction constant was evaluated in each case.     

Isolation of a cDNA from Saccharomyces cerevisiae that encodes a high affinity sulphate transporter at the plasma membrane

Resistance to selenate and chromate, toxic analogues of sulphate, was used to isolate a mutant of Saccharomyces cerevisiae deficient in the capacity to transport sulphate into the cells. A clone which complements this mutation was isolated from a cDNA library prepared from S. cerevisiae poly(A)⁺ RNA. This clone contains an insert which is 2775 by in length and has a single open reading frame that encodes a 859 amino acid polypeptide with a molecular mass of 96 kDa. Sequence motifs within the deduced amino acid sequence of this cDNA (SUL1) show homology with conserved areas of sulphate transport proteins from other organisms. Sequence analysis predicts the position of 12 putative membrane spanning domains in SUL1. When the cDNA for SUL1 was expressed in S. cerevisiae, a high affinity sulphate uptake activity (K_m = 7.5 ± 0.6 M for SO ₄ ^2– ) was observed. A genomic mutant of S. cerevisiae in which 1096 by were deleted from the SUL1 coding region was constructed. This mutant was unable to grow on media containing less than 5 mM sulphate unless complemented with a plasmid containing the SUL1 cDNA. We conclude that the SUL1 cDNA encodes a S. cerevisiae high affinity sulphate transporter that is responsible for the transfer of sulphate across the plasma membrane from the external medium.