光合细菌的分离、保存及其同化磷能力试验研究

为了研究光合细菌的保藏方法及其同化磷能力,采用液体种室内自然放置、液体种蜡封和穿刺物蜡封等方法对光合细菌进行保藏方法研究,结果表明,液体种蜡封和穿刺物蜡封可作为该菌种的长期保存方法;对两株菌的磷同化能力进行了比较,降解率分别为22%和15%。本研究为获得光合细菌的长期保存方法及其在污水净化中的应用提供参考。     

细胞固定化技术在含氮废水处理中的应用研究

生物固定化技术是现代生物工程领域中的一项新兴技术。简要介绍了生物固定化技术,较为系统地阐述了单独包埋和混合包埋两种具体固定化工艺技术及其在处理氨氮废水中应用效果,展望了固定化技术在含氮废水处理中的应用前景与研究发展动向。     

Treating Urban Wastewater: Nutrient Removal by Using Immobilized Green Algae in Batch Cultures

Essential roles of microalgae during the tertiary treatment of municipal wastewater have been proven. In order to avoid wash out of the biomass and also modify the quality of the treated wastewater; some techniques such as cell immobilization have been developed. To do so, in this study four samples from two species of microalgae (Chlorella vulgaris and Chlamydomonas sp.) were determined and confirmed by taxonomic identification. The samples were immobilized in calcium alginate beads. Within 10 days the amounts of nitrate (No₃^?-N) and orthophosphate (Po₄^3?-P) were measured to calculate the removal efficacy for each individual sample. Based on the standard methods, the amount of nitrate and orthophosphate were determined every 3 days within 10 days. The results indicate that immobilized microalgae are able to remove the nutrients and reduce the amount of nitrate and orthophosphate, significantly. Furthermore, the C. vulgaris (YG02) was the best species in this experience with 72% and 99% of reduction in the amount of nitrate and orthophosphate, respectively.     

Biological Synthesis of Semiconductor Zinc Sulfide Nanoparticles by Immobilized Rhodobacter sphaeroides

A novel, clean biological transformation reaction by immobilized Rhodobacter sphaeroides has been developed for the synthesis of zinc sulfide (ZnS) nanoparticles with an average diameter of 8 nm. The nanoparticles were examined by X-ray diffraction, transmission electron microscopy, energy dispersive analyses of X-rays, UV–vis optical absorption and photoluminescence spectra. The average diameter of ZnS nanoparticles varied according to the culture time.     

类球红细菌应用进展

类球红细菌(Rhodobacter Sphaeroides)属光合细菌,是目前研究最深入的光合微生物之一,具有多种代谢方式。不仅能够产生类胡萝卜素、辅酶Q10、超氧化物歧化酶、5-氨基乙酰丙酸和氢气,而且能够降解农药残留、有机废水和多环芳烃等有毒有害物质,应用领域十分广泛。本文综述了类球红细菌在食品、医药、农业、环境、氢气生产等领域中的应用进展,并对类球红细菌的应用前景进行展望。     

Characteristics of light-harvesting complex II mutant of <Emphasis Type="Italic">Rhodobacter sphaeroides</Emphasis> with alterations at the transmembrane helices of β-subunit

The peripheral light-harvesting complex II (LHII) is an important component of the photosynthetic apparatus of Rhodobacter sphaeroides. In this study, genetic, biochemical, and spectroscopic approaches were applied to investigate the spectral properties and functions of LHII in which two amino acid residues Phe32 and Leu42 in the transmembrane helix domain of pucB-encoded β-apoprotein were replaced by Leu and Pro. The mutated LHII complex showed blue shift of absorbance peaks in the near infrared region at ∼801–845 nm in R. sphaeroides. It should be noted that the B800 peak was much lower than that of the native LHII, and transfer energy was efficient from the B800 to the B850 pigments in the LHII complex. The results suggest that the mutated pucB could be expressed in R. sphaeroides, and the functional LHII was assembled into the membrane of R. sphaeroides notwithstanding with the different spectral properties. These mutated residues were indeed critical for the modulation of characteristics and function of LHII complex. Published in Russian in Biokhimiya, 2009, Vol. 74, No. 7, pp. 993–999.     

Simultaneous removal of chemical oxygen demand and nitrate in aerobic treatment of sewage wastewater using an immobilized photosynthetic bacterium of porous ceramic plates

A denitrifying photosynthetic bacterium (PSB), Rhodobacter sphaeroides S was immobilized on a porous ceramic plate (10×10×0.5cm) using a small amount of agar. In the bioreactor (3.5 l liquid) using this plate, repeated batch treatments of sewage wastewater (acetic acid as a carbon source) were carried out under aerobic conditions (DO 5mg/l) for two weeks. The simultaneous and effective removal of chemical oxygen demand (COD), ammonium, and nitrate was observed. In continuous treatment at a relatively high dilution rate of 0.20 h^–1 (retention time, five hours), the same simultaneous treatments were observed for about one month.     

Utilization of volatile fatty acids from the anaerobic digestion liquor of sewage sludge for 5-aminolevulinic acid production by photosynthetic bacteria

Using volatile fatty acids (VFA) from the anaerobic digestion liquor of sewage sludge, up to 9.2 mm 5-aminolevulinic acid (ALA) could be produced by Rhodobacter sphaeroides under anaerobic-light (5 kLux) conditions with repeated addition of levulinic acid (LA) and glycine and using a large inoculum (approx. 2 g/l of cells, initially from glutamate/malate medium). As the VFA medium also contained organic nitrogen sources such as glutamic acid, the cells were later grown up to about 2 g/l in the VFA medium instead of the glutamate/malate medium. ALA production was then again promoted by adding LA and glycine. Using this improved method, up to 9.3 mm ALA was produced by feeding propionate and acetate together with LA and glycine, indicating that VFA medium formed from sewage sludge could be useful for ALA production.     

遮光处理对草珊瑚光合特性及叶片解剖结构的影响

为探索遮光处理对草珊瑚光合特性及叶片解剖结构的影响,为草珊瑚的科学栽培提供理论依据。该研究以2年生草珊瑚为材料,设置全光照(L_0)及50%(L_(50))、70%(L_(70))和90%(L_(90))遮光处理,测定分析其光合特性和叶片解剖结构以及植株生长的变化。结果表明:(1)随着遮光率的增加,草珊瑚叶面光合有效辐射(PAR)、气孔导度(G_s)和蒸腾速率(T_r)逐渐减小,胞间二氧化碳浓度(C_i)先减少后增加,净光合速率(P_n)先增加后减少,并在L_(50)和L_(70)处理下具有较低的PAR、G_s、T_r和C_i,光合‘午休’现象得到有效缓解,表现出更高的P_n。(2)随着遮光率的增加,草珊瑚光补偿点(LCP)逐渐下降,光饱和点(LSP)、暗呼吸速率(R_d)先升高后下降,表观量子效率(AQE)逐渐上升,叶片最大净光合速率(P_(nmax))先升高后下降,并在L_(50)和L_(70)处理下具有较低的LCP,较高的LSP和AQE,以及更高的最大净光合速率。(3)随着遮光率的增加,草珊瑚叶片、表皮细胞、栅栏组织和海绵组织的厚度明显增加,角质层厚度逐渐变薄;在L_(50)和L_(70)处理下,草珊瑚叶片栅栏组织和海绵组织发达,栅栏组织细胞长度变短,海绵组织厚度显著增加,细胞排列疏松,叶绿体数量显著增加;叶肉细胞和叶绿体结构完整,基粒类囊体和基质类囊体片层结构发达,叶片捕获光能力增强。(4)随着遮光率的增加,草珊瑚株高、地径、总叶面积数及单株生物量先增加后减少,并在L_(70)处理下均达到峰值,分别为58.58 cm、5.09 mm、1 038.53 cm~2和15.50 g。研究认为,草珊瑚具有一定的弱光耐受力和光可塑性,50%～70%遮光处理有利于草珊瑚生长,在实际生产中可根据该光照范围选择合适的间套作林分或进行适当的遮光处理。     

Effects of Precise Deletions in Rhodobacter sphaeroides Reaction Center Genes on Steady-state Levels of Reaction Center Proteins: A Revised Model for Reaction Center Assembly

Possible interactions between photosynthetic reaction center (RC) proteins that protect these membrane proteins from proteolytic digestion in RC complex assembly were evaluated by use of translationally in-frame (nonpolar) RC gene-specific deletions. The RC H, RC M and RC L proteins were produced from plasmids, either alone or in concert with one or both of the others, in a strain of Rhodobacter sphaeroides that contained chromosomal deletions of all three RC genes. The steady-state amounts of these proteins in cell membrane and soluble fractions were assessed in western blots. The data are used to propose a model of RC assembly in which the RC M protein accumulates in the cell membrane regardless of the presence of the RC H and RC L proteins, and the RC M protein is a nucleus for addition of RC L followed by RC H in assembly of the RC holocomplex.     

Decolourization and Detoxification of Methyl Red by Aerobic Bacteria from A Wastewater Treatment Plant

Bacterial cultures from a wastewater treatment plant degraded a toxic azo dye (methyl red) by decolourization. Complete decolourization using a mixed-culture was achieved at pH 6, 30 °C within 6 h at 5 mg/l methyl red concentration, and 16 h at 20—30 mg/l. Four bacterial species were isolated that were capable of growth on methyl red as the sole carbon source, and two were identified, namely Vibrio logei and Pseudomonas nitroreducens. The Vibrio species showed the highest methyl red degradation activity at the optimum conditions of pH 6--7, and 30—35 °C. Analysis by NMR showed that previously reported degradation products 2-aminobenzoic acid and N,N-dimethyl-1,4-phenylenediamine were not observed. The decolourized dye was not toxic to a monkey kidney cell line (COS-7) at a concentration of 250 μM. This revised version was published online in July 2006 with corrections to the Cover Date.     

Effects of Extraction of the H-Subunit from Rhodobacter sphaeroides Reaction Centers on Relaxation Processes Associated with Charge Separation

Effects of extraction of the H-subunit from Rhodobacter sphaeroides photosynthetic reaction centers (RC) on the characteristics of the photoinduced conformational transition associated with electron transfer between photoactive bacterio-chlorophyll and primary quinone acceptor were studied. Extraction of the H-subunit (i.e., the subunit that is not directly bound to electron transfer cofactors) was found to have a significant effect on the dynamic properties of the protein–pigment complex of the RC, the effect being mediated by modification of parameters of the relaxation processes associated with charge separation.     

Bioremediation of cadmium by growing Rhodobacter sphaeroides: kinetic characteristic and mechanism studies

The removal kinetic characteristic and mechanism of cadmium by growing Rhodobacter sphaeroides were investigated. The removal data were fitted to the second-order equation, with a correlation coefficient, R2=0.9790-0.9916. Furthermore, it was found that the removal mechanism of cadmium was predominantly governed by bioprecipitation as cadmium sulfide with biosorption contributing to a minor extent. Also, the results revealed that the activities of cysteine desulfhydrase in strains grown in the presence of 10 and 20 mg/l of cadmium were higher than in the control, while the activities in the presence of 30 and 40 mg/l of cadmium were lower than in the control. Content analysis of subcellular fractionation showed that cadmium was mostly removed and transformed by precipitation on the cell wall.     

Biosorption and Bioreduction of Trivalent Aurum by Photosynthetic Bacteria <Emphasis Type="Italic">Rhodobacter capsulatus</Emphasis>

Biosorption has been shown to be an eco-friendly approach to remove heavy metal ions. In this study, the photosynthetic bacteria Rhodobacter capsulatus was screened and found to have strong ability to adsorb Au(III). The maximum specific uptake of living cells was over 92.43 mg HAuCl₄/g dry weight of cell in the logarithmic phase. Biosorpion ability would be enhanced by an acidic environment. As the main cations, during biosorption the quantity of Mg²⁺ exchanged was more than Na⁺. Biosorbed Au(III) could be reduced by carotenoid and enzymes embedded and/or excreted by R. capsulatus, which might be the mechanism of photosynthtic bacteria metal tolerance.     

一组优势菌对焦化废水中吲哚、吡啶的降解条件的实验研究

从普通变形杆菌BH、芽孢杆菌DC4 5、巨大芽孢杆菌F3、枯草芽孢杆菌DC4 2等 4种脱色菌中 ,经驯化、筛选出普通变形杆菌BH、芽孢杆菌DC4 5两株降解苯酚、吲哚、吡啶、喹啉的优势菌。其中普通变形杆菌BH降解吡啶的适宜条件是 :温度 30～ 4 5℃ ,pH 7～ 8,投加适量Pb2 + ;芽孢杆菌DC4 5降解吲哚的适宜条件是 :温度 2 0～ 5 0℃ ,pH 6～ 8,投加适量Zn2 + ,Fe2 + 。     

遮荫对月季光合特性及生长发育的影响

采用人工遮荫法研究不同程度遮荫(75%、50%和25%全光照,以全光照为对照)处理对月季的生长发育、光合特性及叶绿素荧光参数的影响。结果表明:(1)随着遮荫程度的增加,月季的叶片厚度、花径、成花率均呈降低趋势,叶绿素含量呈增加趋势;净光合速率(Pn)、蒸腾速率(Tr)、气孔导度(Gs)、胞间CO2浓度(Ci)降低,气孔限制值(Ls)升高,Pn下降的主要原因可能为气孔限制。(2)月季在遮荫条件下的光饱和点和光补偿点、暗呼吸速率下降,表现出其对光照降低的生理适应机制;其光饱和点在600~1 200μmol·m-2·s-1,光补偿点在29.89~62.95μmol·m-2·s-1,CO2补偿点在78.16~89.41μmol·mol-1,CO2饱和点在1 100μmol·mol-1左右,潜在光合能力13.06~25.63μmol·m-2·s-1,其对光的适应范围较宽,光照和CO2浓度对其光合能力的影响较大。(3)各处理条件下叶绿素荧光参数显示,月季在自然光和低光下(50%和25%全光照)叶片PSⅡ受到了明显伤害,光合作用原初反应过程受到抑制,光合电子传递受到影响。研究发现,月季虽是阳生植物,但光照生态幅较宽,轻度遮荫(75%全光照)能促进其光合作用效率,减少光抑制,植株长势好,成花率高。     

1株水产饵料用光合细菌粪红假单胞菌的分离筛选

从凡纳滨对虾池塘水样、底泥中分离光合细菌,依次通过菌株培养液性状、菌体粗蛋白含量、类胡萝卜素、氨基酸及辅酶Q种类和含量综合评价,筛选营养成分较丰富的菌株,为鱼虾贝类提供优良的开口饵料。试验共分离得到35株光合细菌,其中11株菌液均一、稳定且颜色鲜艳;9株菌粗蛋白含量高于60%;3株菌Rh16、Rh25、Rh34类胡萝卜素含量显著高于其他菌株(P0.05),必需氨基酸与总氨基酸的比值分别为41%、41%和40%,含有的泛醌种类为Q_(10),其中Rh16的Q_(10)含量最高,达6.49μg/mL,可以开发为水产养殖动物优良开口饵料。经培养形态、生理生化特征、16S rRNA基因序列鉴定及系统发育分析表明,菌株Rh16为粪红假单胞菌(Rhodopseudomonas faecalis)。     

类球红细菌类胡萝卜素抗氧化活性研究

研究了研磨法、超声波法、酸溶辅助超声波法和菌体浓度对类球红细菌类胡萝卜素抗氧化活性的影响。结果表明,不同提取方法和固液比条件下,类球红细菌类胡萝卜素均具有清除DPPH自由基能力、抗脂质过氧化能力和还原能力。酸溶辅助超声波法提取的类胡萝卜素产率最高、抗氧化活性最好。类球红细菌类胡萝卜素具有一定的抗氧化活性,其抗氧化活性随菌体浓度的增加而增加。     

Comparative Analyses of Three-Dimensional Models of Bacterial Reaction Centers

Despite the fact that the three-dimensional structure of an integral membrane protein was first determined 20 years ago, structures have been solved for very few membrane proteins. The reaction center is an exception with many mutant and modified structures available from 3 different bacterial species. In order to relate these structures to the function of the reaction center, an accurate assessment of the reliability of the structural models is required. Here we describe the quality of the structures of the bacterial reaction center based upon different criteria, such as evaluation of the geometry of the models and comparison of different models. Overall, the structures are found to be most accurate in the membrane-embedded regions with the periplasmic and cytoplasmic exposed regions having more disorder and differences among the structural models. In general, the cofactors and the surrounding protein regions are among the most accurately determined regions of the protein, except for the secondary quinone and its binding pocket that shows a large variation among structures. The limited accuracy of the secondary quinone is due to its partial occupancy as a consequence of its functional role and to the presence of surface features, including lipids and detergent molecules.     

Metabolically Engineered <Emphasis Type="Italic">Rhodobacter sphaeroides</Emphasis> RV strains for Improved Biohydrogen Photoproduction Combined with Disposal of Food Wastes

Three differently metabolically engineered strains, 2 single PHA^- and Hup^- mutants and one double PHA^-/Hup^- mutant, of the purple nonsulfur photosynthetic bacterium Rhodobacter sphaeroides RV, were constructed to improve a light-driven biohydrogen production process combined with the disposal of solid food wastes. These phenotypes were designed to abolish, singly or in combination, the competition of H₂ photoproduction with polyhydroxyalkanoate (PHA) accumulation by inactivating PHA synthase activity, and with H₂ recycling by abolishing the uptake hydrogenase enzyme. The performance of these mutants was compared with that of the wild-type strain in laboratory tests carried out in continuously fed photobioreactors using as substrates both synthetic media containing lactic acid and media from the acidogenic fermentation of actual fruit and vegetable wastes, containing mainly lactic acid, smaller amounts of acetic acia, and traces of higher volatile acids. With the lactic acid-based synthetic medium, the single Hup^- and the double PHA^-/Hup^- mutants, but not the single PHA^- mutant, exhibited increased rates of H₂ photoproduction, about one third higher than that of the wild-type strain. With the food-waste-derived growth medium, only the single Hup^- mutant showed higher rates of H₂ production, but all 3 mutants sustained a longer-term H₂ photoproduction phase than the wild-type strain, with the double mutant exhibiting overall the largest amount of H₂ evolved. This work demonstrates the feasibility of single and multiple gene engineering of microorganisms to redirect their metabolism for improving H₂ photoproduction using actual waste-derived substrates.