厌氧产氢菌的分离筛选、诱变选育及其产氢特性的研究

本文从四川绵竹酒厂、成都市龙泉长安垃圾填埋场以及四川大学荷花池底的厌氧污泥中先后分离得到63株厌氧产氢菌，其中H-8、H-61、HC-10等16株产氢细菌产氢能力较高，HC-10的产氢能力最高，最大产氢量和最大产氢速率分别达到2840 ml H2/L培养基和25.39 mmol H2/g drycell·h，对HC-10进行生理生化鉴定和分子生物学鉴定，判定其为clostridium sp.，对HC-10的产氢条件进行了研究，结果表明，该菌的最适生长温度为35 ℃，最适生长初始pH为7，以葡萄糖为最佳碳源，以蛋白胨为最佳氮源，不利用无机氮源，其产氢发酵液相产物以乙醇和乙酸为主，其发酵类型属于乙醇型发酵。此外，以酒糟废液作为底物，进行了菌株HC-10的生物强化试验，研究表明，投加了HC-10的强化系统其产氢量比对照高出40.32%。 同时为了获得厌氧产氢菌的高效突变株，分别以产氢菌H-8和H-61为原始菌株进行微波诱变处理，对微波诱变参数进行了优化，考察了突变株的遗传稳定性、产氢特性及耐酸性。菌株H-8经过微波诱变得到5株高产氢突变株HW7、HW33、HW181、HW184、HW195，经多次传代表明HW195是稳定的高产突变株。突变株HW195具有较好的耐酸性，在pH值为2.8时仍能生长。通过间歇发酵实验，其最大产氢量和最大产氢速率分别达到2460 mL/L培养基和27.97 mmol H2/g drycell·h，比原始菌分别提高了50.75%和41.7%。菌株H-61经过微波诱变后选育得到的突变株HW-18，其最大产氢量和最大产氢速率分别达到2190 mL/L培养基和25.86 mmol H2/g drycell·h，比原始菌分别提高了23.03%和31.00%。 为了对比各种诱变方式对产氢菌产氢能力的影响，以厌氧产氢菌H-61为原始菌株，先后经亚硝基胍(NTG)、紫外(UV)诱变，选育得到1株高产突变株HCM-23。在葡萄糖浓度为10 g/L的条件下，其产氢量为3024 mL/L培养基，比原始菌株提高了69.89%；其最大产氢速率为33.19 mmol H2/g drycell·h，比原始菌株提高了68.14%。经过多次传代实验，稳定性良好。其发酵末端产物以乙醇和乙酸为主，属于典型乙醇型发酵。其最适产氢初始pH为6.5，最适生长温度为36 ℃，以蔗糖为最佳碳源。与原始菌株相比，突变株HCM-23的产氢特性发生了改变，如生长延滞期延长，可利用无机氮源等。 From anaerobic activated sludge, 16 strains of hydrogen producing bacteria were newly isolated. One of them named as HC-10 had the highest hydrogen producing capability, under the batch fermentative hydrogen production condition, the maximal hydrogen yield and hydrogen production rate was 2840 mL/L culture and 25.39 mmol H2/g drycell·h. It was identified as clostridium sp.HC-10 by 16S rDNA sequence analysis. Various parameters for hydrogen production, including substrates, initial pH and temperature, have been studied. The optimum condition for hydrogen producing of strain HC-10 were achieved as: initial pH 7.0, temperature 35 ℃, glucose as the favorite substrate, Moreover, using distiller's solubles wastewater as substrate, HC-10 strain was added in the biohydrogen producing system to research the bioaugmentation effection. The results showed that the hydrogen production of bioaugmentation system was 40.32% higher than the noaugmentation system. An anaerobic, hydrogen producing strain H-8 was irradiated by microwave to optimize the microwave mutagenesis condition, and to test the heredity, hydrogen-producing potential and aciduric of the mutants. An aciduric mutant named as HW195 with steady hydrogen-producing capability was obtained, which can grow at pH 2.8. Its capability of hydrogen production was tested in the batch culture experiments. The maximum hydrogen yield and hydrogen production rate was 2460 mL/L culture and 29.97 mmol H2/g drycell·h, which was 50.7% and 41.7% higher than those of the initial strain, respectively. When used the strain H-61 as original strain, a mutant named as HW18 was obtained. The maximum hydrogen yield and hydrogen production rate was 2190 mL/L culture and 25.86 mmol H2/g drycell·h, which was 23.03% and 31.00% higher than those of the initial strain, respectively. The results demonstrated that microwave mutagenesis could be used in the field of hydrogen producing microorganism. The hydrogen producing strain H-61 was used as an original strain which was induced by NTG and UV for increasing and the hydrogen production capability. One of the highest efficient H2-producing mutants was named as HCM-23 with its stable hydrogen production capability. which was tested in the batch culture experiments. With the condition of 10 g/L glucose, its cumulative hydrogen yield and hydrogen production rate was 3024 mL/L culture and 33.19 mmol H2/g drycell·h, 69.89％and 68.14％ higher than that of the original strain, respectively. The terminal liquid product compositions showed that the mutant HCM-23 fermentation was ethanol type, while the original strain H-61 fermentation was butyric acid type. Varieties of parameters of hydrogen production fermentation were studied, including time, carbon source, nitrogen source, glucose concentration, glucose utilization, initial pH and incubation temperature had been studied, indicated the optimum condition of hydrogen production for the mutant HCM-23 as initial pH6.5, temperature 36 ℃, and the favorite substrate was sucrose. The hydrogen production characters of the mutant and the original strain were different, such as, the growth lag phase and the utilization of inorganic nitrogen source, etc. This work shows a good application potential of NTG-UV combined mutation in the biohydrogen production. And the hydrogen production mechanism and metabolic pathway should be explored furthermore.

一株高温硫酸盐还原菌的分离、鉴定及其对金属腐蚀研究

从四川化工总厂热交换器中分离到一株嗜热硫酸盐还原菌，该菌对金属有腐蚀作用。菌体形状呈微弯或直杆状，大小为0.5-0.8μm宽，2.8-4.3μm长，革兰氏阴性，周生鞭毛，运动，形成次端生的椭圆形芽孢，细胞内部构造丰富，菌落在不含Fe++盐的培养基上生长时，呈灰白色，直径为0.5mm，边缘不整齐，在含Fe++盐的培养基上生长时则为黑色，直径为2-3mm，菌落中央有0.5mm左右的小突起，该菌最适生长温度为50℃，最适PH6.5，不耐盐，可于沸水(99℃)中处理6小时而不失活，能利用H2＋CO2，乙醇，卜葡糖 ，可溶性淀粉，甲酸盐，乳酸盐，丙酮酸盐为唯一碳源，生长需要酵母膏，无脱硫弧菌素和细胞色素C3。

耐酸产甲烷菌的分离及其复合菌剂的研究

本文从成都龙泉垃圾填埋场和宜宾造纸厂分离到耐酸性能优良的高温产甲烷菌RY3和中温产甲烷菌SH4，并将其与实验室现有的利用不同底物的产甲烷菌配伍组合成了复合菌剂。采用活性污泥作为固体附着物，研制出了固体产甲烷菌复合菌剂。 菌株RY3的pH耐受范围为5.5～10.5，最适生长pH 6.0～8.0。菌株RY3为革兰氏阳性，长杆状，多数单生，不运动；菌落浅黄色，形状近圆形；利用H2+CO2或甲酸盐作为唯一碳源生长，不利用乙酸盐，对氯霉素非常敏感。该菌最适生长温度为55℃～65℃，最适NaCl浓度为0～2%。根据形态和生理生化特性及16S rDNA序列分析将其初步定为热自养甲烷热杆菌（Methanothermobacter thermautotrophicus）。添加RY3菌液与仅添加厌氧污泥作为接种物相比一周内可使达到最大产甲烷速率所需时间缩短三分之二，甲烷总产量提高约1.8倍。菌株SH4的生长pH范围5.5～9.5，其对酸碱具有良好的适应性，培养3天后，在初始pH值为6.0～8.0的培养基中甲烷产量相差不大，且基本达到最大产量。SH4革兰氏染色阳性，短杆状，多数单生，不运动；菌落近圆形，微黄；利用H2+CO2或甲酸盐作为唯一碳源生长，不利用乙酸盐，对氯霉素非常敏感。SH4最适生长pH 为7.0，最适生长温度为35℃，最适NaCl浓度为0～1.5%。实验表明，添加SH4菌液与仅添加厌氧污泥作为接种物相比可使产甲烷启动时间缩短三分之一，甲烷总产量亦有大幅提高。从形态和生理生化特征以及16S rDNA序列分析表明SH4为嗜树木甲烷短杆菌（Methanobrevibacter arboriphilus）。 以活性污泥为附着物，与培养基和菌种经搅拌后厌氧发酵可得产甲烷菌固体复合菌剂。固体复合菌剂的pH耐受范围为5.5～9.5，温度耐受范围为15℃～65℃，表明其对环境的适应性较强。以猪粪为底物进行厌氧发酵，接种复合菌剂进行试验，以接种实验室长期富集的产甲烷厌氧污泥作为对照，在20℃时，发酵甲烷浓度与对照基本一致，但每日产气量优于对照，第15天时接种复合菌剂的发酵瓶每日产气量是对照的1.59倍；50℃时达到最大甲烷含量所需时间比对照缩短三分之二，三周内总产气量约为对照的2.7倍，甲烷总产量约为2.8倍。以不加接种物为对照，接种复合菌剂20℃时发酵甲烷含量达到50%约需2周，对照2周内甲烷含量最高仅为4.3%；50℃时接种复合菌剂发酵仅需约1周甲烷含量便可达50%，对照则至少需要2周。 In this paper, high-temperature Methanogen RY3 and middle-temperature SH4 were isolated from Chengdu Longquan refuse landfill and Yibin paper mill. They could be used to make compound inoculum that producing methane with the existing Methanogens utilized different substrate. With using anaerobic activated sludge be solid fixture, the process had been designed to produce solid compound inoculum. Strain RY3 possessed excellent capacity of acid and alkali-tolerant. The pH-tolerant scale of RY3 was 5.5～10.5 and its optimum pH value for growth was 6.0～8.0. RY3 was G+, long-rod shape, monothetic and nonmotile, the colony was pale yellow with suborbicular-shape. Formate or H2+CO2 but not acetate was utilized by RY3 as sole C-source, and it was very sensitive to chloramphenicol. Besides, strain RY3 grew fastest at 55℃～65 and 0℃～2% NaCl. Characteristics of modality and physiology with sequence analysis of the 16s rDNA gene of strain RY3 preliminarily showed that it was Methanothermobacter thermautotrophicus. The experiments indicated that the time which began to produce methane with the highest velocity could be shortened two third by adding RY3 in one week, and the total methane production also was 1.8 times than before. Strain SH4 possessed wide scale of growing pH（5.5～9.5）and excellent ability of acclimatizing itself to acid-alkali. The methane production had no apparent difference among those cultivated in different initial pH（6.0～8.0）after three days and equaled to the maximum production basically. Cells of SH4 were G+, short-rod sharp, monothetic and nonmotile. The colony was pale yellow with suborbicular-shape. Formate or H2+CO2 but not acetate was utilized by SH4 as sole C-source, and it was very sensitive to chloramphenicol. Besides, it grew fastest at pH 7.0，55 ℃～65 and 0℃～2% NaCl concentration. The experiment indicated the time that began to produce methane could be shortening one third by adding SH4. And the total methane production also rose apparently. Characteristic of modality and physiology with sequence analysis of the 16S rDNA gene of strain SH4 demonstrated it was Methanobrevibacter arboriphilus. The activated sludge was utilized as fixture, mixed with culture medium and inocolum, that the solid compound inoculum could be produced by anaerobic fermentation. The compound inoculum could grow between pH 5.5～9.5, 15℃～65. It demonstrated the compound inoculum ha℃ve great ability of adapting to circumstance. In the experiment that making pig manure be substrate and taking the anaerobic sludge producing methane that cultured in long term in laboratory to be comparison, the concentration of methane in fermentation added compound inoculum almost equal to the comparison at 20℃, but the volume of gas production could be a little higher. The gas production everyday inoculated compound inoculum was 1.59 times to comparison. The time that the concentration of methane to maximum could be shortening by two third by adding compound inoculum, and the total gas production was 2.7 times to comprison while the total methane production was 2.8 times. If take the no inoculum be the comprasion, anaerobic fermentation added compound inoculum made the concentration of methane to 50% in 2 weeks but the comparison only to 4.3% at 20℃. The time that the concentration of methane to 50% by adding compound inoculum only need 1 week, but the comparison need 2 weeks at 50℃.

CsI(T1)晶体探测器APD读出的温度效应

对中国科学院近代物理研究所自行生长的铊激活碘化铯闪烁晶体CsI(T1)光输出及Hamamtsu公司生产的S8664-1010型雪崩光二极管(APD)增益对温度的依赖关系做了系统研究.结果表明,CsI(T1)晶体光产额在室温范围内随着温度的增加而增加,在-2℃—8℃温度范围内的平均温度系数为0.67%/℃,在8℃—25℃温度范围内的平均温度系数为0.33%/℃.而对所使用的APD,其增益在室温范围内的温度系数为-3.68%/℃(工作电压400V).APD结合CsI(T1)晶体在室温下对~(137)Cs的662keYγ射线的能量分辨可达5.1%.

河流阶地的形成及其对构造与气候的意义

IEECAS SKLLQG

28.7MeV/u的~(14,16,18)O+~(7,9)Be反应中的同位素分布研究

用同位旋相关的Boltzmann Langevin方程研究了在入射能量为2 8 7MeV/u下 ,不同弹核 14O ,16 O和 18O轰击不同靶核 7Be和 9Be的反应 ,计算了生成碎片的产生截面 ,发现用丰中子 (缺中子 )炮弹或丰中子 (缺中子 )靶进行反应 ,所得到的产物均有丰中子 (缺中子 )的碎片出现 .同位素分布宽度和峰位与入射体系密切相关 ,产生碎片的电荷数越接近入射弹核的电荷数 ,则同位素分布的宽度越大 ,峰位偏离β稳定线值越远 ,其同位旋效应越明显 .

同位旋相关的Boltzmann-Langevin方程及新核素~(19)Na产生截面的研究

介绍了一种同位旋相关的输运方程 ,研究了在入射能量为 2 8.7和 6 0 .0MeV/u时　 12 C +12 C的反应 ,对模型进行检验 ,发现计算结果较好地符合实验结果 ,说明了方程的可靠性 .利用该模型研究了在入射能量为 2 8.7MeV/u下反应系统17— 2 0 ,2 2 Ne+12 C中核素19Na的产生截面 ,发现缺中子核引起的反应 ,具有更大19Na的产生截面 ,为新核素的探测找到了理论依据 .

兰州放射性核束流线的磁铁与电源

:描述了兰州重离子加速器国家实验室最近建成的兰州放射性核束流线(RIBLL)的磁铁和电源系统的物理设计和性能特点。考虑高阶分量的影响,二级偏转磁铁的入口和出口极面采用二次曲面形状,除使磁铁起偏转作用外,还有聚焦和发散的功能。RIBLL中首次采用大功率开关电源,并获得成功

The experimental measurement of the relative biological effectiveness of carbon ions with different qualities

The relative biological effectiveness (RBE) of carbon ions with linear energy transfer (LET) of 172 keV/mu m and 13.7 keV/mu m were determined in this study. The clonogenic survival and premature terminal differentiation were measured on normal human. broblasts AG01522C and NHDF after exposure of the cells to 250 kV X-rays and carbon ions with different qualities. RBE was determined for these two biological end points. The results showed that the measured RBE10 with a survival fraction of 10% was 3.2 for LET 172 keV/mu m, and 1.33 for LET 13.7 keV/mu m carbon ions. RBE for a doubling of post-mitotic. broblasts (PMF) in the population was 2.8 for LET 172 keV/mu m, and 1 for LET 13.7 keV/mu m carbon ions. For the carbon ion therapy, a high RBE value on the Bragg peak results in a high biological dose on the tumour. The tumour cells can be killed effectively. At the same time, the dose on healthy tissue would be reduced accordingly. This will lighten the late effect such as fibrosis on normal tissue.

Progresses of heavy-ion cancer therapy

The status of heavy-ion cancer therapy has been reviewed. The existing and constructing heavy-ion beam facilities for cancer therapy in the world are introduced. The first clinical trials of superficially placed tumor therapy at heavy ion research facility in Lanzhou (HIRFL) are presented.

Separation of potential and kinetic electron emission from Si and W induced by multiply charged neon and argon ions

T he total secondary electron emission yields, gamma(T), induced by impact of the fast ions Neq+ (q = 2-8) and Arq+ (q = 3-12) on Si and Neq+ (q = 2-8) on W targets have been measured. It was observed that for a given impact energy, gamma(T) increases with the charge of projectile ion. By plotting gamma(T) as a function of the total potential energy of the respective ion, true kinetic and potential electron yields have been obtained. Potential electron yield was proportional to the total potential energy of the projectile ion. However, decrease in potential electron yield with increasing kinetic energy of Neq+ impact on Si and W was observed. This decrease in potential electron yield with kinetic energy of the ion was more pronounced for the projectile ions having higher charge states. Moreover, kinetic electron yield to energy-loss ratio for various ion-target combinations was calculated and results were in good agreement with semi-empirical model for kinetic electron emission.

eta production off the proton in a Regge-plus-chiral quark approach

A chiral constituent quark model approach, embodying s- and u-channel exchanges, complemented with a Reggeized treatment for the t channel is presented. A model is obtained allowing data for pi(-)p ->eta n and gamma p ->eta p to be described satisfactorily. For the latter reaction, recently released data by the CLAS and CBELSA/TAPS Collaborations in the system total energy range 1.6 less than or similar to W less than or similar to 2.8 GeV are well reproduced by the inclusion of Reggeized trajectories instead of simple. and. poles. The contribution from "missing" resonances, with masses below 2 GeV, is found to be negligible in the considered processes.

Role of five-quark components in radiative and strong decays of the Lambda(1405) resonance

Within an extended chiral constituent quark model, the three- and five-quark structure of the S-01 resonance Lambda(1405) is investigated. Helicity amplitudes for electromagnetic decays [Lambda(1405)->Lambda(1116)gamma, Sigma(1194)gamma] and transition amplitudes for strong decays [Lambda(1405)->Sigma(1194)pi, K- p] are derived, as well as the relevant decay widths. The experimental value for the strong decay width, Gamma(Lambda(1405)->(Sigma pi)degrees) = 50 +/- 2MeV, is well reproduced with about 50% of a five-quark admixture in the Lambda(1405). Important effects owing to the configuration mixing among Lambda P-2(1)A, Lambda P-2(8)M, and Lambda P-4(8)M are found. In addition, transitions between the three- and the five-quark components in the baryons turn out to be significant in both radiative and strong decays of the Lambda(1405) resonance.

形变核179Pt在束γ谱学和重丰中子新核素的合成及其衰变性质研究

本论文分为两部分，分别沿自旋和同位旋自由度开展研究工作。第一部分介绍原子核高自旋态研究的相关背景知识、基础理论等，描述在束γ谱学研究的实验技术和方法，分析和讨论形变核179Pt转动带能级结构的特性，并采用相关理论对其进行分析。通过融合蒸发反应149sm（35cl，P4n）布居奇A核179Pt的高自旋激发态。指认179Pt的组态为1／2-［521］，5／2-［512〕和7／2＋〔633〕的三条转动带。在hco＝0．27 Mev附近，观测到1／2-〔521］带内顺排角动量突然增大，建议该现象是由一对113／2中子发生顺排造成的。另外观察到在7／2＋〔633］带中出现较大的旋称劈裂。建议由于三轴形变加强了波函数中来自几＝1／2轨道的组分，从而导致较大的旋称劈裂的出现。论文第二部分介绍重丰中子核衰变性质研究以及新核素合成、鉴别工作的相关背景知识、理论基础及实验技术。并对相关的研究工作分别进行阐述。通过快中子反应合成并鉴别了新核素19705和新的同核异能素186rnTa，利用γ（x）谱学方法首次建立了19705的部分衰变纲图。测得它们的半衰期分别为2.8士0.6 min和1.50.1 min，并与理论计算结果进行了比较；进行了原子核基态β延发裂变（pDF）的实验研究，经测量首次发现了230Ac的两个pDF事件，测得23OAc的pDF几率为(1.19±0.40)

高电荷态离子与固体表面作用的离子溅射和电子发射的实验研究

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