Submerged culture of Magne-tospirillum gryphiswaldense under N2-fixing condition and regulation of activity of nitrogen fixation

A submerged culture technique for Magnetospirillum gryphiswaldense under the nitrogen-fixing condition (microaerobic and N-limited) was set up. In N-limited medium with Na-lactate as a sole carbon source, the optical density (A600 nm) and activity of nitrogen fixation of cells were 1.3 and 217 nmol of ethylene produced per hour per A600nm respectively within 21 h by three times of feeds. The pH and temperature were controlled at 7.2 and 30℃ respectively, and the oxygen concentration was controlled by sparging with N2 containing 0.4%-0.8% of O2. The activity of nitrogen fixation of cells was obviously inhibited by oxygen and ammonium. It indicated that the posttranslational regulation of nitrogenase existed in M. gryphiswaldense.     

油樟叶精油不同馏分化学成分及其抗氧化能力评价

采用减压分馏的方法对油樟叶精油进行分馏,结合GC-MS联用技术研究油樟叶精油及其不同馏分的化学成分,并对其清除1,1-二苯-2-三硝基苯肼(DPPH)自由基能力进行了评价。结果表明:油樟叶精油及其不同馏分中的化学成分及其含量差异较显著。从油樟叶精油(A_0号精油)及其所得到的馏分(A_1号精油、A_2号精油、A_3号精油)中分别鉴定出29、15、20和31种物质,共鉴定出31种化合物,包括烃类21种、醇类8种、酮类和酯类各1种,它们的共有成分有15种,此外还检测到一些特有成分,油樟叶精油(A_0号精油)14种、馏分A_2号精油5种、馏分A_3号精油16种。油樟叶精油及其不同馏分均有清除DPPH自由基能力,它们的抗氧化活性能力强弱顺序为A_3 A_2 A_0 A_1。但它们清除能力均低于同质量浓度的维生素C。本研究结果为油樟叶精油有效成分的进一步分离纯化及其资源开发利用可以提供理论依据。     

抗艾滋病病毒蛋白CVNH在大肠杆菌中表达条件的优化

CVNH(Cyanovirin-N homology)蛋白家族是具有抗HIV活性的蓝藻抗病毒蛋白-N(Cyanovirin-N,CVN)的同源物。在前期研究中,已首次获得水蕨Ceratopteris thalictroides CVNH基因的全长序列,构建了pET32a-CtCVNH的原核表达载体。以此为基础,对CVNH在大肠杆菌Rosetta 2(DE3)中的表达条件进行优化。分别探讨了最佳培养基类型及成分、培养基初始pH、诱导时机、诱导剂浓度及诱导时间。优化后的诱导条件是:含24 g.L-1酵母提取物和72 g.L-1胰蛋白胨的TB培养基、培养基初始pH为8.0、菌液A600nm为0.6时加入1.6 g.L-1乳糖诱导6 h。CVNH蛋白的表达量较优化前提高了1.9倍。这为进一步开展CVNH的药物研制和开发奠定基础。     

Cloning and functional analysis of the sequences flanking mini-Tn5 in the magnetosome-deleted mutant NM21 of Magnetospirillum gryphiswaldense MSR-1

A magnetosome-deleted mutant NM21 of Magnetospirillum gryphiswaldense MSR-1 was generated by mini-Tn5 lacZ2 transposon mutagenesis, and a 3073-bp fragment flanking mini-Tn5 lacZ2 in NM21 was cloned by Anchored PCR. Sequencing analysis showed that this fragment involved three putative ORFs; the mini-Tn5 lacZ2 was inserted into ORF1. Functional complementary test indicated that the 3073-bp fragment was required for biosynthesis of magnetosomes in M. gryphiswaldense MSR-1. The majority of proteins, which had homology with the protein encoded by ORF1, were the cation transporter. Transmembrane domain analysis showed that the protein encoded by ORF1 contained four transmembrane domains. It may be a transmembrane protein. The protein encoded by ORF1 contained two putative conserved domains: COG0053 and PRK09509. The MMT1 and FieF, containing conserved domains COG0053 and PRK09509 too, were Fe²⁺ transporter (cation diffusion facilitator superfamily). It was suggested that the protein encoded by ORF1 might take part in the magnetosomes biosynthesis as Fe²⁺ transporter. Supported by National Natural Science Foundation of China (Grant No. 30570023) and Scientific Research Project of Huaibei City, Anhui Province (Grant No. 070114)     

The synthesis and characterization of a Co-N/C composite catalyst for the oxygen reduction reaction in acidic solution

A non-precious metal Co-N/C catalyst for the oxygen reduction reaction (ORR) was synthesized by heating a mechanical mixture of cobalt chloride, urea and acetylene black under a nitrogen atmosphere. The catalyst was characterized by XRD and XPS. The electrocatalytic activity in the ORR was evaluated by linear sweep voltammetry in 0.5 mol L⁻¹ H₂SO₄ solution. The results show that the Co-N/C catalyst aids the reduction of oxygen. The presence of elemental cobalt in the precursor allows nitrogen atoms to embed themselves in the graphite matrix to form pyridinic and graphitic type C-N structures as the ORR active sites. The effect of heat-treating temperature on the catalytic activity was also investigated. The results also show that the Co-N/C catalyst is most active when pyrolyzed at 600°C. The obtained Co-N/C catalyst loses some activity after initial exposure to the H₂SO₄ solution because of leaching, but is then stable for up to 20 h immersion. The catalyst is also stable when charged, which is supported by the cyclic voltammetry results.     

Carbon-nitrogen/graphene composite as metal-free electrocatalyst for the oxygen reduction reaction

Sheet-like carbon-nitrogen (CNx)/graphene composites with a high content of nitrogen (x≤0.15) was prepared by the carbonization of polypyrrole (PPy)/reduced-graphene-oxide (rGO) composite at 600-800°C. We used rGO instead of graphene oxide (GO) sheets as a template and a substrate to immobilize PPy since the PPy/GO composite agglomerates easily because of the dehydration of excess oxygen-containing groups on the GO sheets during the drying process. The dried PPy/rGO intermediate and its derived CNx/graphene products retain their high dispersion and loose-powder features. The as-prepared CNx/graphene composites have a total nitrogen content of about 10 at% and their nitrogen state is mainly of pyridinic and graphitic type. CNx/graphene composites exhibit excellent performance for the oxygen reduction reaction (ORR) in terms of electrocatalytic activity, stability and immunity towards methanol crossover and CO poisoning, suggesting their potential as metal-free electrocatalysts for the ORR.     

Construction and characterization of partiallyntrC-deleted mutants inAlcaligenes faecalis

To study the effect of ntrC gene product on the expression and regulation of other important nitrogen-fixing genes in Alcaligenes faecalis, partially ntrC-deleted mutants of A. faecalis have been generated. To start with, the ntrC gene of A. faecalis was cloned into a suicide plasmid pSUP202 to create a recombinant plasmid pSUM1. The ntrC gene in pSUM1 was then replaced by a lacZ-Km^r fragment resulted in the generation of a plasmid pSUM2. The lacZ fragment in pSUM2 was further removed and a plasmid pSUM3 produced. As a second step, the plasmid pSUM2 or pSUM3 was introduced into the wild type of A. faecalis A1501 by conjugation and two partially ntrC-deleted mutants A15CM1 (ntrC∷lacZ) and A15CM2 (ntrC ^- ) were obtained. To understand the regulatory effect of the NtrC on the expression of nifH and nifA, a nifH-lacZ gene or a nifH-lacZ gene was introduced into the ntrC^- mutant by conjugation. The results indicated that: (ⅰ) although the ntrC^-mutant was nif ⁺ , its nitrogen fixation activity was only 20% that of the wild type; (ⅱ) the ntrC^- mutant failed to grow on the medium containing nitrate as a sole nitrogen source; (ⅲ) the regulation of ntrC gene expression did not require its own product; (ⅳ) the expression of nifH in A . faecalis was positively regulated by the ntrC. Deletion of the ntrC resulted in the reduction of nifH expression or even totally inactivated nitrogen fixation; (ⅴ) there was no obvious influence on the expression of nifA in A. faecalis if the ntrC gene was deleted.     

Synthesis, crystal structure and thermal behavior of Rb(NTO) · H2O

The Rb(NTO)·H₂O crystal has been synthesized by reaction of 3-nitro-1,2,4-triazole-5-one (NTO) with Rb₂CO₃ in aqueous solution. Its crystal structure has been determined. The crystal belongs to monoclinic system. Crystal structure data: space group P2₁ / n; a = 0.633 0(1), b = 0.824 1(2), c =1.296 4(3) nm; β= 97.90(1)°; V = 0.669 9 (2) nm³ , Z = 4, D_c = 2.306 g/cm³, μ= 7.365 nm^-1, F (000) = 448. An eight coordinated compound is formed between Rb⁺ with oxygen atoms and nitrogen atoms. A layer structure is formed by coordination bonds and hydrogen bonds. The thermal decomposition mechanism of this coordination compound is discussed .     

Preparation and catalytic performance of Fe-SBA-15 and FeO x /SBA-15 for toluene combustion

High-surface-area and well-ordered mesopor- ous Fe-incorporated SBA-15 （xFe-SBA-15） and SBA-15- supported FeOx （yFeOx/SBA-15） with the Fe surface den- sity between 0.09 to 1.11 Fe-atom/nm2 have been prepared using the one-step synthesis and incipient wetness impregnation methods, respectively. Physicochemical properties of these materials were characterized by means of numerous techniques, and their catalytic activities for the combustion of toluene were evaluated. It is found that the xFe-SBA-15 and yFeOx/SBA-15 samples possessed rod- or chain-like morphologies. The Fe species were of high dispersion when the Fe surface density was lower than 0.76 Fe-atom/nm2 in xFe-SBA-15 and 0.64 Fe-atom/nm2 in yFeOx/SBA-15. At a similar Fe surface density and space velocity, the xFe-SBA-15 catalysts showed better activity than the yFeOJSBA-15 catalysts, in which the xFe-SBA- 15 catalyst with Fe surface density 0.59 Fe-atom/nm2 performed the best. It is concluded that the good perfor- mance of the xFe-SBA-15 sample with Fe surface density 0.59 Fe-atom/nm2 was associated with its large surface area, high Fe species dispersion, and good low-temperature reducibility.     

3种脱蛋白方法对虎斑乌贼肌肉多糖体外抗氧化活性的影响

【目的】比较3种不同脱蛋白方法对虎斑乌贼肌肉多糖体外抗氧化活性的影响。【方法】采用Sevag法、三氯乙酸法和等电点法对经过热水浸提的虎斑乌贼肌肉多糖进行脱蛋白,同时对脱蛋白多糖的体外抗氧化活性进行研究。【结果】3种不同脱蛋白方法得到的多糖均具有一定的抗氧化能力。当多糖浓度为5 mg/mL时,Sevag法、三氯乙酸法和等电点法得到的脱蛋白多糖对1,1-二苯基-2-三硝基苯肼(DPPH)自由基的清除率分别为87.05%、42.35%和38.54%,相应的IC50值分别为1.96mg/mL、11.81mg/mL和9.84mg/mL;对羟基自由基(·OH)清除率分别为93.67%、70.80%和68.75%,相应的IC50值分别为0.67mg/mL、1.37mg/mL和2.72mg/mL;对超氧阴离子(O-2)清除率分别为72.33%、62.42%和53.33%,相应的IC50值分别为0.15mg/mL、1.56mg/mL和3.81mg/mL。多糖还原力大小分别为0.183,0.160和0.144。【结论】3种脱蛋白方法所得虎斑乌贼肌肉多糖抗氧化活性大小依次为Sevag法三氯乙酸法等电点法。     

轮叶黑藻腐解过程营养物质释放及微生物胞外酶活性变化规律

通过轮叶黑藻腐解模拟试验,分析了底泥和砂石2种基质条件下营养物质释放和微生物胞外酶活性变化特征。结果表明,试验结束时底泥组和砂石组的轮叶黑藻质量损失率分别为64. 62%和62. 15%。植物腐解过程释放的营养物质使得水体溶解氧含量迅速降低,TN、TP质量浓度迅速增加。试验结束时(146 d),上覆水体的水质基本恢复初始水平。水体有机质在56 d以色氨基酸类蛋白质为主,之后逐渐以类富里酸和类腐殖酸类物质为主。植物腐解过程与微生物胞外酶活性具有一定的响应关系,跟水体水质及植物本身的特性相关。纤维素酶、蔗糖酶密切相关(r=0. 70,P0. 05),且受植物TOC和水体TN的影响,过氧化氢酶与水体的DO和EC相关(P0. 05),碱性磷酸酶及脲酶则分别受TP和NH_3-N的抑制调节(P0. 05)。     

The induction of Sinorhizobium meliloti C4-dicarboxylate transport system （Dct） is regulated by oxygen concentration

The Sinorhizobium meliloti C4-dicarboxylate transport （Dct） system is essential for symbiotic nitrogen fixation. The dctA gene, encoding the C4-dicarboxylate permease, is expressed in both free living and symbiotic cells. But in free living cells expression of dctD and dctB is absolutely required for the expression of dctA. In this study, in order to investigate the effect of oxygen concentration on the induction of Dct system, E. coli DH5a strain which carries the plasmid-encoded dctABD operon was used in tube assays. It was found that the specific induction of Dct system oc- curred only at a certain depth under the surface of M63-0.6% agar media, suggesting that Dct system could respond to oxygen concentration during succinate-induced expression. Furthermore, when measured at different oxygen concentrations, the highest expression level was observed at oxygen concentration of 2%. Thus, we predict that in addition to dicarboxylates, the induction of Dct system may also regulated by oxygen concentration.     

Specific degradation of photosystem II D1 protein by a protease （Air3815） in heterocysts of the cyanobacterium Anabaena sp. PCC7120

Some filamentous cyanobacteria form heterocysts under conditions lacking combined nitrogen for nitrogen fixation.Photosystem II is removed from heterocyst during the process of cell differentiation.Here,we demonstrate that Alr3815 is a protease that is capable of degrading D1 protein of photosystem II.Strain-322,which lacks alr3815,is impaired in nitrogen fixation in air because some oxygen evolving activity is retained in its heterocysts.Our results also suggest that calcium may play a regulatory role in D1 degradation during heterocyst differentiation.     

不同氮素形态肥料对油茶春梢生长的影响

为了给油茶林合理施肥管理提供科学依据,本研究以13 a生"岑软3号"油茶(Camellia oleifera)树为研究对象,以不施肥作为空白对照,以不加氮肥作为无氮对照,研究相同施氮量下硝态氮肥、铵态氮肥、酰胺态氮肥以及3种形态氮素1:1:1混合态氮肥对油茶春梢形态及生理的影响.结果显示,与空白对照相比,混合态氮对春梢...     

响应面法优化醋酸钙不动杆菌菌株23的脂肪酶产酶条件

采用响应面法对醋酸钙不动杆菌(Acinetobacter calcoaceticus)菌株23的发酵产脂肪酶培养基进行优化实验。实验首先考察该菌株产酶所需的最适碳源和氮源,然后用Plackett-Burman法设计实验确定影响产酶的主要影响因素,用最陡爬坡实验逼近最大响应区域,最后通过Box-Behnken方法进行二次回归分析得到最佳的产酶条件为:营养肉汤18.4%、蛋白胨9.18%、吐温800.52%、K2HPO40.2%,橄榄油1%,MgSO40.05%,CaCl20.05%,FeSO40.1%。在该优化条件下,发酵液的脂肪酶活力可以达到15.2U/ml,比优化前提高3倍。     

Visible light photocatalytic decoloration of methylene blue on novel N-doped TiO2

Novel N-doped TiO2 (denoted as N-NTA600) was prepared by treating nanotube titanic acid (NTA) in NH3 flow. Its visible light photocatalytic activity,evaluated by decoloration reaction of methylene blue,is higher than that of N-P25(600) prepared by treatment of P25-TiO2 in the same condition. It is suggested that the origin of visible-light photocatalytic activity is single-electron-trapped oxygen vacancy (Vo·) modified by chemisorbed NO.     

Synthesis, crystal structure and properties of binuclear manganese complex [(bipy)2Mn2(μ-O)(μ-Ac)2(H2O)2](ClO4)2

Binuclear manganese complex [(bipy)₂Mn₂(μ-O)(μ-Ac)₂(H₂O)₂](ClO₄)₂ was synthesized by the reaction of MnAc₃ · 2H₂O with 2,2′-bipyridine in the HAc-NaAc buffer (pH = 4.0). X-ray diffraction result for the single crystal shows that the crystal is monoclinic, space group C₂/C, with a = 3.408 2(7),b = 0.864 4(2),c = 2.174 9(4) nm, β= 105.2∘, V=6.186(2) nm³, Z= 8. There are two very strong peaks of UV-Vis spectrum in the range of 400–600 nm, which are similar to those of Mn catalase and Mn ribonuleotide reductase extracted from organisms. Cyclic voltammogram shows that the complex in CH₃CN undergoes quasi-reversible one-electron reduction and oxidation at E_1/2=1.15V.