荧光素酶基因luc在大肠杆菌中表达条件优化

对重组荧光素酶大肠杆菌菌株M15／pQE30-luc进行了表达条件的优化研究。单因素结果表明：在初始pH值7．0,装液量为20％,2％的接种量,终浓度为0．5mmol／L的IPTG,添加10—30mmol／L的Mg^2＋,摇床转速为200r／min,37℃诱导3．5h酶的表达量最高。正交试验结果表明：初始pH值为7．0,添加40mmol／LMg^2=,接种量2％,装液量为20％时表达量最高,比酶活达1．63×10^8RFU／mg蛋白。     

球衣菌对工业废水中Cu2+吸附条件的研究

利用球衣菌FQ32制备生物吸附剂,研究其对Cu^2＋的耐受能力,并以连江某工厂含Cu^2＋废水为样液,对其进行吸附条件试验,分析了影响吸附率和吸附量的因素,结果表明,球衣菌FQ32对Cu^2＋的耐受能力大于150mg／L。在工业废水中Cu^2＋的浓度为23．1mg／L,吸附剂用量为0．2g／L,30℃,pH值6．0,吸附时间10min的条件下,该吸附剂对Cu^2＋的吸附率达到85．2％,吸附量达91．21mg／g。     

微紫青霉菌菌株GXCR对低品位黄铜矿中的Cu和Fe的生物淋滤及其淋滤机制

研究了Penicillium janthinellum菌株GXCR对低品位黄铜矿中的Cu和Fe的生物淋滤浸出.结果表明摇浸淋滤效率优于静置浸没淋滤效率,对Cu的淋滤浸出效果最佳;在添加最佳碳源(10%蔗糖,W/V)、氮源(1.5% NaNO3,W/V)、摇浸淋滤和最佳条件组合(淋滤培养基初始pH 6.0,矿石大小200目,矿石浓度5%(W/V)和初始接种菌量3.0×105分生孢子/mL)时,Cu的浸出率达到87.31%(W/W);摇浸淋滤时影响Cu的生物浸出的主要因素是淋滤培养基初始pH(FF0.05);对Cu和Fe淋滤起主要作用的有机酸分别是柠檬酸和草酸;浸没淋滤效率低是与柠檬酸和草酸产量低有关;GXCR的生物淋滤机制有2种:柠檬酸和草酸的生化作用和菌体附着生长所产生机械压力对矿石的破碎作用.     

零价铁对2,4-二氯酚生物还原脱氯的影响研究

采用间歇试验,接种驯化两月的厌氧混合微生物,考察厌氧体系中添加零价铁（Fe^0）对2,4-二氯酚（2,4-DCP）生物还原脱氯效果的影响,并对影响“Fe^O＋微生物”体系的一些因素进行了探索。结果显示：与零价铁或微生物的单独作用相比,“Fe^O＋微生物”体系能够有效促进2,4-DCP的脱氯反应,最佳Fe^O投加量和微生物接种量分别为0.5g／L和376.2mgVSS／L;初始pH=8.0对2,4-DCP的转化效果最好,偏酸性环境不利于污染物转化;微生物接种量与铁用量之间有一适宜比例,一定范围内增加微生物接种量可催生出更多可降解污染物的酶或酶系,提高2,4-DCP的降解效果。     

红螺菌对Cu2+的吸附研究

研究了4株红螺菌(R-01,R-02,R-03,R-04)对Cu^2 的生物吸附行为。结果表明,3株红螺菌(R-01,R-02,R-04)对20mg／L的Cu^2 有较高的吸附率,其中R-04达99．1％。进一步研究了R-04菌体的最佳吸附条件,在pH2、浓度为80mg／L Cu^2 、35℃、微光厌氧下吸附45min,吸附率、吸附量分别达94％,48．08mg／g。在一定的浓度范围内红螺菌对Cu^2 的吸附符合Langmuir吸附模型和Freundlich吸附模型,但符合Langmuir吸附模型的程度更优。     

中等嗜热细菌浸出黄铜矿过程中pH对群落结构的影响

为了优化浸出工艺,研究了pH对浸矿过程主要微生物种群结构的影响。用中度嗜热混合菌槽浸黄铜矿精矿,在不控制pH,控制pH为2．5及控制pH为1．2时,应用PCR-RFLP（限制性酶切片段长度多态性）方法对上述浸出条件下的细菌群落动态变化进行研究。结果表明,浸出体系只有两种微生物,一种为Acidithiobacillus Caldus,一种为Leptospirillum ferriphilum。pH对群落结构有明显影响。不控制pH时,浸出开始阶段At．caldus是优势种群,占群落的96％,随着浸出的进行,L．ferriphilum增多,在浸出后期代替At．caldus成为优势菌种,占69％。控制pH时,L．ferriphilum始终占主导地位,同时发现pH为2．5时At．caldus在群落中的丰度比pH为1．2时高。     

细菌浸出高硫锰矿及菱锰矿的试验

本报告用氧化亚铁硫杆菌(Thiobacillus ferrooxidans)把硫酸亚铁氧化产生酸性硫酸高铁溶液。在Fcs+浓度25克／升,pH1．8,矿石粒度120目,矿浆浓度7％,浸矿温度60℃条件下搅拌浸出高硫锰矿及菱锰矿,2．5和4小时锰浸出率分别达82％及98％。硫锰矿石中除硅酸锰不能浸出外,矿石中可溶浸的锰矿物浸出了99．2％。硫酸高铁在程矿反应时全部水解为氧氧化铁沉淀,不能矬续循环使用。但以氢氧化铁吸收高硫锰矿焙烧脱硫时排出的含低浓度二氧化硫烟道废气,使还原为亚铁。亚铁液经加温等处理后,由新接种的细菌氧化再生成硫酸高铁,反髭浸矿,如此循环。     

黄孢原毛皮革菌降解苯胺的工艺研究

通过正交试验优化筛选了适合黄孢原毛皮革菌降解苯胺的适宜培养基和摇瓶培养降解条件。结果表明：其适宜降解的液体培养基组成为：蔗糖20g／L,可溶性淀粉20g／L,(NH4)2SO4l0g／L,Mn^2 lμmol／L,Tween-800．3％,蛋白胨30g／L。适宜降解的摇瓶培养条件为：接种量为20％、pH为7．0、温度为30℃、培养时间为12d．此条件下的苯胺最高降解率可达95．5％。     

维纳斯捕蝇草及其叶片离体快繁技术研究

捕蝇草是一种喜欢生活在潮湿环境,叶片能捕捉小虫子的有趣植物。本试验尝试用叶片做外植体,不经过愈伤组织阶段,直接诱导出丛生芽,缩短培养时间。比较了附加不同种类和浓度激素的培养基对诱导不定芽生成的影响。实验证明,诱导丛生芽较适宜培养基：1／2MS＋6-BA2．0mg／L＋肉汁10％＋活性炭0．2％：继代扩繁适宜培养基：1／2MS＋6-BA1．0mg／L＋NAA0．3mg／L＋肉汁10％＋活性炭0．2％：最佳生根培养基：1／2MS＋NAA0．3mg／L＋肉汁10％＋活性炭0．2％。     

木霉LaTr 01菌株产漆酶发酵的条件

从华南地区采集土样,采用愈创木酚平板筛选产漆酶菌株,获得了一株短周期产漆酶的小型丝状真菌。通过观察菌落特征、生长情况以及显微镜下菌丝和孢子的形态,初步鉴定该菌株为木霉属的一个种（Trichodermaspp、）,命名为木霉LaTr01菌株。通过单因素方法研究该菌产漆酶的发酵条件,结果表明：LaTr01的产酶培养基以麦芽糖为最佳碳源;以酵母提取物为最适氮源;培养24h后加入Cu“比培养开始加入Cu ^2＋LaTr01产酶活高出约1倍。采用麦芽糖、酵母提取物、Cu^2＋浓度L9（3^3）的正交试验优化漆酶发酵条件,结果表明,氮源是影响该菌产漆酶的最重要因素,碳源次之,Cu^＋浓度影响较小;LaTr01菌株产生漆酶的最佳条件为：5g／L酵母提取物、20g/L麦芽糖、1．5mmol／LCu^2＋,Cu^2＋加入时间为培养24h后。在优化的培养条件下,该菌酶活可达480．556U／L。     

Column Bioleaching of Low-Grade Chalcopyritic Ore Using Moderate Thermophile Bacteria

Bacterially assisted heap leaching is an economical technology for treating low grade copper sulphides. In the present research, bioleaching of low grade chalcopyrite ore (1% chalcopyrite, and 3% pyrite) have been investigated using moderate thermophilic bacteria. The ore sample has low solubility in acid solution (about 5%). Experiments were carried out using column reactors and the effect of particle size (?12.7, ?19.07 and ?25.04 mm) and external addition of carbon dioxide to the induced air (10% v/v) have been investigated. Results have shown that the copper recovery increased with reducing particle size, and carbon dioxide addition improved bacterial activity and copper dissolution. In the optimum condition, i.e., particles finer than 12.07 mm and 10% (v/v) carbon dioxide addition, 69.68% of copper were extracted.     

The effects of Fe(II) and Fe(III) concentration and initial pH on microbial leaching of low-grade sphalerite ore in a column reactor

In this study the effects of initial concentration of Fe(II) and Fe(III) ions as well as initial pH on the bioleaching of a low-grade sphalerite ore in a leaching column over a period of 120 days with and without bacteria were investigated. Four different modifications of medium were used as column feed solutions to investigate the effects of initial concentration of Fe(II) and Fe(III) ions on zinc extraction. The experiments were carried out using a bench-scale, column leaching reactor, which was inoculated with mesophilic iron oxidizing bacteria, Acidithiobacillus ferrooxidans, initially isolated from the Sarcheshmeh chalcopyrite concentrate (Kerman, Iran). The effluent solutions were periodically analyzed for Zn, total Fe, Fe(II) and Fe(III) concentrations as well as pH values. Bacterial population was measured in the solution (free cells). Maximum zinc recovery in the column was achieved about 76% using medium free of initial ferrous ion and 11.4 g/L of ferric ion (medium 2) at pH 1.5. The extent of leaching of sphalerite ore with bacteria was significantly higher than that without bacteria (control) in the presence of ferrous ions. Fe(III) had a strong influence in zinc extraction, and did not adversely affect the growth of the bacteria population.     

国内某低品位难选铜矿的生物浸出研究

目的:微生物湿法冶金技术是一种有效回收难处理常规选矿方法难以处理的复杂矿中金属的方法,本研究旨在利用该工艺处理国内某低品位(0.67%)难选铜矿,提高铜的回收率。方法:首先,从某矿山富集得到中温富集物,其次,对该矿石进行生物浸出,同时优化浸出过程工艺参数。结果:所富集得到的中温富集物最适生长温度为30℃,最适pH值为1.9。在摇瓶中浸出难选铜矿时,最佳摇床转速为180 r/min,最优充气强度为360 mL/min,10天内难选铜矿中铜的浸出率可以达到92%。结论:该中温富集物具有较好地浸出难选铜矿的能力。     

Bioleaching of a Spanish uranium ore

Abstract: The bioleaching of a low-grade uranium ore in shaker and columns using natural, pure and mixed cultures has been studied. Initially, a chemical and microscopical characterization of the mineral was carried out. Orbital shaker experiments were performed to quickly obtain the best bacterial leaching conditions. Afterwards, small columns were used to determine other variables related to percolation leaching. Finally, an uranium ore from the F6 mine (Ciudad Rodrigo, Spain) was leached in the presence of bacteria using large columns (24 cm diameter and 275 cm height). The most important results were: (i) the ore contains sufficient pyrite in order for uranium bioleaching to take place under optimum conditions; (ii) shaker experiments showed that temperature, pH and type of inoculum are the most important variables in bioleaching; (iii) two different ores were attacked in the columns: altered and non-altered. In the first case, the extraction rate of uranium was higher. However, in both cases the final efficiency was very similar (95%).     

Optimization of Microbial Leaching of Base Metals from a South African Sulfidic Nickel Ore Concentrate by Acidithiobacillus ferrooxidans

X-ray diffraction analysis revealed that pentlandite and chalcopyrite were the prominent mineral phases in a South African sulfidic nickel ore concentrate that hosted nickel and copper. Cobalt was found to be closely associated with the nickel-bearing pentlandite phase of the ore sample. Microbial batch leaching experiments designed according to a central composite design model were run for 15 days in a shaking incubator (150 rpm) at a constant temperature (30°C) with variations in experimental parameters like ore pulp density, particle size, bacterial inoculum, pH of the culture medium, and residence time. Quadratic mathematical models were developed to predict the rate of metal extractions. The suitability of the model of the microbial leaching process was confirmed from normal probability curves. An analysis of variance indicated that the residence time, pulp density of the ore, and particle size were the most significant factors. Bacterial inoculum size hardly showed any effect on the total metal extractions. Maximum nickel (82%), cobalt (76%) and copper (25.6%) extractions were achieved under optimum conditions, operated for 15 days at pulp density of 2% and particle size of ?75 µm at pH 1.5.     

Bioleaching of copper from chalcopyrite ore by fungi

Microorganisms have been geologically active in mineral formation, mineral diagenesis and sedimentation via direct action of their enzymes or indirectly through chemical action of their metabolic products. This property of microorganisms is being harnessed during the recent years for extraction of metals from their ores, especially from low-grade ores. In the present study bioleaching of copper from its low-grade chalcopyrite ore using 26 isolates of acidophilic fungi is reported. Most of these fungal strains belonged to the genera Aspergillus, Penicillium and Rhizopus. The leaching experiments were conducted in Czepek Dox minimal medium containing 1% (100 mesh) ore with shaking at room temperature for 20 days. Out of these, 4 isolates exhibited significant bioleaching activities. Maximum leaching of copper (78 mg/L) was observed with Aspergillus flavus (DSF-8) and Aspergillus niger (DOF-1). Nutritional and environmental conditions for optimum bioleaching were standardized. Present study indicates the usefulness of acidophilic fungi in bioleaching of copper from its low-grade ores.     

Comparison of bioleaching behaviors of different compositional sphalerite using <Emphasis Type="Italic">Leptospirillum ferriphilum</Emphasis>, <Emphasis Type="Italic">Acidithiobacillus ferrooxidans</Emphasis> and <Emphasis Type="Italic">Acidithiobacillus caldus</Emphasis>

Two sphalerite samples with different iron/sulphur (Fe/S) ratios, Shuikousan ore (Fe/S 0.2) and Dachang ore (Fe/S 0.52), were processed using three microbial species, Leptospirillum ferriphilum, Acidithiobacillus ferrooxidans and Acidithiobacillus caldus. Following 20 days of bioleaching in shake flask cultures, a higher zinc (Zn) extraction (96%) was achieved with Shuikousan ore than with Dachange ore (72%). The extraction efficiency increased when elemental S was added to Dachang ore to attain the same Fe/S ratio as that for Shuikousan ore. Following the addition of S, the redox potential, pH and total dissolved Fe for Dachang ore demonstrated similar behaviors to those of Shuikousan ore. Acidithiobacillus caldus and L. ferriphilum became the dominant species during the bioleaching of sphalerite with a high Fe/S ratio. In contrast, the dominant species were A. ferrooxidans and A. caldus during the bioleaching of sphalerite with a low Fe/S ratio. These results show that the Fe/S ratio has a significant influence on the bioleaching behavior of sphalerite and the composition of the microbial community.     

响应面法优化Fe~(2+)的微生物氧化条件

氧化亚铁钩端螺旋菌(Leptospirillum ferrooxidans,L.f)是一种极端嗜酸,专性自养氧化铁的细菌,能够耐受较低pH和较高的温度,被广泛应用于生物浸矿和环境治理。氧化亚铁钩端螺旋体菌的生物浸矿效率与其对Fe~(2+)氧化速率相关,因此,本文采用响应面法,通过建立二次多项式回归方程考察pH、温度、Fe~(2+)浓度及转速四个培养因素对Fe~(2+)氧化速率的影响。结果显示在pH为2.25、温度为32℃、初始Fe~(2+)浓度为175.36 mmol/L、转数为165 r/min时,Fe~(2+)最高氧化速率为0.2911 g/Lh。     

内切中性纤维素酶EGⅡ的发酵条件优化

对毕赤酵母基因工程菌EIM-50-eg2产内切中性纤维素酶的主要影响因子进行研究,考察氮源、pH、温度、微量元素PTM1和甲醇浓度等对工程菌产酶的影响。单因素实验和正交实验结果表明,优化后的培养基组成及培养条件:磷酸氢二铵40 g/L,甲醇15 mL/L,硫酸镁10 g/L,磷酸二氢钾9 g/L,初始pH 6.0,培养温度28℃,PTM1添加量0.02%,甲醇诱导浓度1.5%。优化后内切葡聚糖酶活力可达4 158 U/(mL.min)是优化前1 449 U/(mL.min)的2.86倍。     

Effect of Activated Carbon Addition on the Conventional and Electrochemical Bioleaching of Chalcopyrite Concentrates

The effect of activated carbon addition on the rate and efficiency of copper mobilization from Sarcheshmeh chalcopyrite concentrate was studied in the presence and absence of a mixed culture of moderately thermophilic microorganisms. Conventional leaching at a 10% (w/v) pulp density in 500-ml Erlenmeyer flasks on a rotary shaker at 150 rpm, and electrochemical bioleaching in a stirred bioreactor at an ORP (oxidation-reduction potential) range of 400 to 430 mV measured against a Ag/AgCl reference electrode. The bioreactor contained ore concentrate at a pulp density of 20%, which was stirred at 600 rpm. All experiments were conducted in the presence and absence of 3 g/L activated carbon, at initial pH 1.5, temperature 50°C, in Norris's nutrient medium with an addition of 0.02% (w/v) yeast extract. The results showed that the addition of activated carbon increased the rate and yield of copper extraction from the concentrate especially in the presence of bacteria. Final recovery after 20 days was 52% and 44% in the shake flask experiments with and without carbon addition, respectively. Enhanced rates of copper mobilization were achieved in the electrochemical bioleaching experiments in which copper was leached selectively relative to iron. Final copper recovery after 10 days was 85% and 77% in the presence and absence of activated carbon, respectively. The positive effect of activated carbon on copper extraction could be related to the galvanic interaction between the inert carbon as cathode and chalcopyrite as anode. The bacterial elimination of sulfur produced on the sulfide minerals during chemical leaching is assumed to intensify the galvanic interaction. It seems that maintaining the ORP at a low potential and efficient mixing improves the bacterial and chemical subsystems in the electro-bioreactor that accelerates the rate of copper mobilization from the concentrate.