嗜酸异养菌对自养菌Acidithiobacillus ferrooxidans金属离子抗性和生物浸出的影响

【目的】了解嗜酸异养菌在诸如酸性矿坑水(AMD)和生物浸出体系等极端酸性环境中对浸矿微生物产生的影响。【方法】研究由嗜酸异养菌Acidiphilium acidophilum和自养菌Acidithiobacillus ferrooxidans经长期驯化后形成的共培养体系分别在Cd2+、Cu2+、Ni2+和Mg2+胁迫下的稳定性;并将此共培养体系应用于黄铁矿和低品位黄铜矿的生物浸出实验。【结果】在上述4种金属离子分别存在的条件下,异养菌Aph.acidophilum均能促进At.ferrooxidans对亚铁的氧化,提高其对能源利用的效率。共培养体系中的异养菌Aph.acidophilum使At.ferrooxidans对Cu2+的最大耐受浓度(MTC)由2.0 g/L提高到5.0 g/L,而且共培养的细胞数量与2.0 g/L Cu2+条件下生长的At.ferrooxidans纯培养相似。另外,共培养中的At.ferrooxidans对Mg2+的MTC也由12.0 g/L提高到17.0 g/L。生物浸出实验中嗜酸异养菌Aph.acidophilum促进了At.ferrooxidans对黄铁矿样品的浸出,浸出率较其纯培养提高了22.7%;但在含铁量较低的低品位黄铜矿浸出体系中共培养和At.ferrooxidans纯培养的浸出率均低于33%。在加入2.0 g/L Fe2+的低品位黄铜矿浸出体系中,共培养和At.ferrooxidans纯培养的浸出率均得到提高,分别达到52.22%和41.27%。【结论】以上结果表明,Aph.acidophilum与At.ferrooxidans共培养在一定的环境胁迫下仍能保持其稳定性并完成各自的生态功能,并且嗜酸异养菌Aph.acidophilum适合在含铁量较高的浸出体系中与铁氧化细菌共同作用来提高生物浸出的效率。     

铜耐受定向驯化强化嗜酸喜温硫杆菌浸出贫黄铜矿

生物浸出贫黄铜矿后期,累积的Cu2+会对微生物产生胁迫效应从而影响浸出效率,因此该类微生物的铜耐受性定向驯化变得非常关键。对嗜酸喜温硫杆菌(Acidithiobacillus caldus)进行6个月铜耐受定向驯化,将出发菌株与驯化菌株在不同铜胁迫浓度下(0、1和3 g/L Cu2+)纯培养及浸出贫黄铜矿,并比较分析关键参数变化。在纯培养体系中,3 g/L Cu2+胁迫下最高比生长速率(μmax)由驯化前0.21 d-1(13 d)提升至0.54 d-1(10 d)。在0、1和3 g/L Cu2+浸出体系中,菌株驯化后铜浸出率分别较出发菌株提高17.64%、70.93%及306.09%。扫描电镜(SEM)分析矿渣形貌差异表明伴随胁迫作用增强菌体对矿物腐蚀程度变弱,在相同铜胁迫水平下,驯化菌株浸出体系矿渣表面呈现更多潜在的吸附位点及明显腐蚀痕迹。采用傅里叶红外光谱(FTIR)分析矿渣表面关键官能团变化,驯化菌株浸出体系的矿渣呈现更多含硫基团。X射线衍射(XRD)分析矿渣成分表明驯化菌株浸出体系中呈现较为丰富的铁硫衍生物如Fe3O4、FeS。综上,驯化后A. caldus具备较强抵御铜胁迫能力,在浸出体系中保持了更活跃的生化浸出效应,有望在类似的工业生物浸出过程中发挥其潜在优势。     

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.     

铜蓝精矿的生物浸出

采用两种嗜酸硫杆菌(嗜酸氧化亚铁硫杆菌和喜温硫杆菌)对铜蓝进行生物浸出,实验在有或没有4 g/L硫酸亚铁pH 2．0、150转/分、35℃的三角瓶中进行。实验结果表明:用两种菌混合浸出的铜几乎等于嗜酸氧化亚铁硫杆菌单独浸出的铜;另外,亚铁的加入能提高铜的浸出。     

阿萨希毛孢子菌通过分泌铁载体在低铁限制性条件下生长

目的探讨阿萨希毛孢子菌在低铁限制性条件下生长的机制,为后续联合常用抗真菌药物治疗播散性毛孢子菌病提供实验室依据。方法 200μmol/L的去铁胺加入酵母浸出粉胨葡萄糖培养基(Yeast Extract Peptone Dextrose Medium,YPD)培养基中制备低铁限制性培养基,检测阿萨希毛孢子菌临床分离株在该培养基中的生长曲线,并检测低铁条件下,阿萨希毛孢子菌产生铁载体的情况。结果阿萨希毛孢子菌的生长曲线不受低铁限制性条件的影响,且其在低铁环境下可以产生铁载体。结论阿萨希毛孢子菌通过产生铁载体在低铁限制性条件下正常生长。     

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。     

氧化亚铁硫杆菌亚铁氧化活性与其对低品位铜矿浸矿速率关系的研究

氧化亚铁硫杆菌是一个具有很强生物浸矿能力的细菌,本文对3株分离得到的氧化亚铁硫杆菌及一株来自菌种中心(Acidithiobacillus ferrooxidans A．f)的铁氧化活性及其这些菌株对低品位黄铜矿浸出速率进行了研究。结果显示,在所有的4株A．f菌中,菌株CMS—F1和F10—ATCCC23270的铁氧化活性较高,其对黄铜矿生物浸出速率也高。进一步分析亚铁氧化活性对生物浸矿效率的影响时发现,在A．f菌中,氧化活性高的菌株,其对低品位黄铜矿的生物浸出效果也高。     

不同碳源对解磷真菌溶解低品位磷矿能力的影响

以磷酸三钙作为唯一磷源,从磷矿区筛选出一株高效浸磷真菌.经过核糖体间隔区(ITS)扩增序列分析,确定该菌株属于黑曲霉.研究不同碳源条件下该菌株对低品住磷矿中磷浸出能力的影响;此外,还利用高效液相色谱法(HPLC)和耦合等离子体-发射光谱仪对溶磷机理进行了初探.结果表明,以可溶性淀粉作为碳源的条件下,真菌对低品位磷矿的磷浸出率最高,可能与其代谢产生的草酸和苹果酸及其有机酸总量有关,同时,磷浸出率与培养介质pH值呈显著负相关.     

生物浸出系统中Acidithiobacillus ferrooxidans对雄黄表面改性研究

【目的】研究Acidithiobacillus ferrooxidans BY-3对雄黄表面改性作用,为进一步研究雄黄的生物炮制技术提供实验基础与理论依据。【方法】在4组生物浸出体系中(每组包含100 mL无亚铁离子的9K培养基和0.500 g雄黄):第1组无添加;第2组添加4.469 g硫酸亚铁;第3组添加0.100 g硫粉;第4组加入4.469 g硫酸亚铁和0.100 g硫粉。在上述4组中使用A.ferrooxidans BY-3对雄黄进行生物浸出。浸出前后雄黄表面形貌及元素变化,使用扫描电镜(SEM)与能谱仪(EDS)、X-射线衍射(XRD)、拉曼光谱(Raman)、电感耦合等离子体原子发射光谱仪(ICP-AES)进行分析。【结果】4组浸出体系均发现A.ferrooxidans BY-3粘附于雄黄表面以此来产生直接作用。含Fe2+的浸出体系中雄黄表面产生非常明显的变化,含硫的浸出体系中雄黄表面变化不明显;只有Fe2+存在的浸出体系中As/S比率增高,而其余3组浸出体系中As/S比率均明显下降;另外,改性雄黄的表面存在黄钾铁矾、硫、赤铁矿、针铁矿和磁铁矿等,但未检测到砷华(As2O3)与副雄黄(Pararealgar)。【结论】A.ferrooxidans对雄黄改性具有重要作用。Fe2+对雄黄的改性具有促进作用,而硫对雄黄的改性具有抑制作用。雄黄改性前后的物化分析结果证实了生物浸出技术可有效解决传统方法制备雄黄及贮存过程中氧化和光化问题。     

功能菌群耦合黄铁矿浸出软锰矿的研究

【目的】将3种不同来源的环境样品混合后接种至含1%黄铁矿和1%软锰矿的培养基中进行富集培养,初步得到有一定浸矿功能的混合微生物菌群。【方法】菌群继续用于黄铁矿和低品位软锰矿共同浸出,设置未接种的体系作为对照。【结果】对浸出过程中菌群结构的变化、pH、锰浸出率和浸出残渣的成分进行分析,结果发现接种过微生物菌群的浸出体系在反应15 d后,锰浸出率达到92.48%,远高于未接菌对照组的40.34%;菌群中Thiomonas sp.所占比例从最初的2%上升到浸出结束时的93%。实验组的pH从最初的4.0下降到2.5;X射线衍射(XRD)分析发现,通过生物作用浸出的残渣中含有黄钾铁矾,说明生物代谢产生了大量的硫酸。【结论】证明微生物在两矿浸出过程中通过促进黄铁矿解离,维持体系低pH等作用加速反应的进行。结果为进一步研究微生物浸矿的作用机制和开发低品位锰矿的生物浸出工艺打下了基础。     

BP神经网络优化微生物浸矿工艺

以低品位黄铜矿溶液为原料,浸矿制备Cu2＋能有效提高低品位黄铜矿的利用价值。基于浸矿过程中存在多因素影响的现象,通过正交试验与神经网络分析方法,对浸矿条件（接种量、矿石品位、Fe2＋添加量及浸矿溶液pH）实行优化。结果表明：在正交试验组中最佳试验结果为浸矿产128．753mg／LCu^2＋;BP神经网络优化后的最佳实验组合为微生物接种量12％、矿石品位0．3％、添加Fe^2＋24g/L及浸矿溶液pH1．7,该条件下验证试验产Cu^2＋ 141．352mg／L,通过正交试验及神经网络优化提高了微生物浸出低品位黄铜矿酸性溶液Cu^2＋的产量。     

Microbial leaching of metals from sulfide minerals

Microorganisms are important in metal recovery from ores, particularly sulfide ores. Copper, zinc, gold, etc. can be recovered from sulfide ores by microbial leaching. Mineral solubilization is achieved both by 'direct (contact) leaching' by bacteria and by 'indirect leaching' by ferric iron (Fe(3+)) that is regenerated from ferrous iron (Fe(2+)) by bacterial oxidation. Thiobacillus ferrooxidans is the most studied organism in microbial leaching, but other iron- or sulfide/sulfur-oxidizing bacteria as well as archaea are potential microbial agents for metal leaching at high temperature or low pH environment. Oxidation of iron or sulfur can be selectively controlled leading to solubilization of desired metals leaving undesired metals (e.g., Fe) behind. Microbial contribution is obvious even in electrochemistry of galvanic interactions between minerals.     

紫外线和60Co辐照诱变选育高效嗜酸氧化亚铁硫杆菌

依次利用紫外线和60Co-γ射线辐照诱变的方法对嗜酸氧化亚铁硫杆菌进行诱变,选育高效嗜酸氧化亚铁硫杆菌菌株.结果表明,紫外诱变可以有效提高嗜酸氧化亚铁硫杆菌的Fe2+氧化速率,最佳紫外线诱变时间为240 s.诱变后菌株的Fe2+氧化速率从0.273 g/L/h提高到了0.312 g/L/h.继续利用60Co-γ射线进行...     

红豆杉高产细胞系快速建立和更新的新方法

目的：通过优化培养基和培养条件,建立太行红豆杉速生细胞系,为红豆杉细胞工厂化生产提供原种细胞系和繁殖技术。方法：以太行红豆杉为材料,筛选B5、WPS、MS等3种培养基和6-卞基嘌呤（6-BA）、萘乙酸（NAA）、2,4-二氯苯氧乙酸（2,4-D）等3种激素组合,采用液体、半固体、纸桥共3种培养方法继代培养并筛选高产细胞系。结果：B5＋NAA（1.0mg/L）＋6-BA（0.5mg/L）＋2,4-D（1.0mg/L）为最适愈伤组织诱导培养基,诱导率达100%;B5＋NAA（1.0～1.5mg/L）＋2,4-D（1.5～2.0mg/L）＋6-BA（0.5mg/L）为最佳继代培养基,生长量最高可达0.30g/（g·d）（鲜重）。结论：初步建立了红豆杉细胞系快速建立和繁殖新方法,用半固体培养产生初代细胞系、液体培养将细胞系同步化、纸桥培养快速繁殖细胞系,并通过检测紫杉醇含量不断更新细胞系。该细胞系体系建立方法简单,纯化速度快,易更新,产物细胞系可作为工厂化生产的原种细胞系。     

Temperature Effects on Bacterial Leaching of Sulfide Minerals in Shake Flask Experiments

The microbiological leaching of a sulfide ore sample was investigated in shake flask experiments. The ore sample contained pyrite, pyrrhotite, pentlandite, sphalerite, and chalcopyrite as the main sulfide minerals. The tests were performed at eight different temperatures in the range of 4 to 37°C. The primary data were used for rate constant calculations, based on kinetic equations underlying two simplified models of leaching, i.e., a shrinking particle model and a shrinking core model. The rate constants thus derived were further used for the calculation of activation energy values for some of the sulfide minerals present in the ore sample. The chalcopyrite leaching rates were strongly influenced by the interaction of temperature, pH, and redox potential. Sphalerite leaching could be explained with the shrinking particle model. The data on pyrrhotite leaching displayed good fit with the shrinking core model. Pyrite leaching was found to agree with the shrinking particle model. Activation energies calculated from the rate of constants suggested that the rate-limiting steps were different for the sulfide minerals examined; they could be attributed to a chemical or biochemical reaction rather than to diffusion control.     

聚磷茵JPA1菌株的生长特性和聚磷条件的优化

以YG培养基为基础,考察不同条件对聚磷菌JPA1菌株生长及聚磷效率的影响。结果表明：JPA1菌株生长的对数期为3-9h,9h后进入稳定期,12h到达衰亡期。在单因素试验中,麦芽糖为JPA1菌株最适生长和聚磷的C源,最适生长温度为30-35℃,pH为8,Na＋、Ca2＋、Mg2＋、Mn2＋和K＋等离子有利于JPA1菌株的生长;最适聚磷温度为30-35℃,pH为7,Fe2＋、Na2＋、Mg2＋、Mn“和K＋等离子有利于JPA1菌株的聚磷。正交试验结果表明：JPA1菌株去磷培养基的最优组合为麦芽糖1．5g／L,温度37℃,pH6．5,装液量50mL（300mL摇瓶）;菌株最适扩增培养基为麦芽糖1．5g/L,温度37℃,pH6．5或7．5,装液量150mL（300mL摇瓶）。     

Kinetics of iron oxidation by Leptospirillum ferriphilum dominated culture at pH below one

The kinetics of ferrous iron oxidation by Leptospirillum ferriphilum (L. ferriphilum) dominated culture was studied in the concentration range of 0.1-20 g Fe(2+)/L and the effect of ferric iron (0-60 g Fe(3+)/L) on Fe(2+) oxidation was investigated at pH below one. Denaturing gradient gel electrophoresis of PCR amplified 16S rRNA genes followed by partial sequencing confirmed that the bacterial community was dominated by L. ferriphilum. In batch assays, Fe(2+) oxidation started without lag phase and the oxidation was completed within 1 to 60 h depending on the initial Fe(2+) concentration. The specific Fe(2+) oxidation rates increased up to around 4 g/L and started to decrease at above 4 g/L. This implies substrate inhibition of Fe(2+) oxidation at higher concentrations. Haldane equation fitted the experimental data reasonably well (R(2) = 0.90). The maximum specific oxidation rate (q(m)) was 2.4 mg/mg VS . h, and the values of the half saturation (K(s)) and self inhibition constants (K(i)) were 413 and 8,650 mg/L, respectively. Fe(2+) oxidation was competitively inhibited by Fe(3+) and the competitive inhibition constant (K(ii)) was 830 mg/L. The time required to reach threshold Fe(2+) concentration was around 1 day and 2.3 days with initial Fe(3+) concentration of 5 and 60 g/L, respectively. The threshold Fe(2+) concentration, below which no further Fe(2+) oxidation occurred, linearly increased with increasing initial Fe(2+) and Fe(3+) concentrations. Fe(2+) oxidation proceeds by L. ferriphilum dominated culture at pH below 1 even in the presence of 60 g Fe(3+)/L. This indicates potential of using and biologically regenerating concentrated Fe(3+) sulfate solutions required, for example, in indirect tank leaching of ore concentrates.     

铁改性沸石对Clostridium acetobutylicum XY16固定效率及发酵性能的影响

考察4种无机铁盐改性沸石对丁醇生产菌Clostridium acetobutylicum XY16的固定效率及其发酵产丁醇性能的影响。结果表明:铁改性沸石对菌体的固定效率均优于未改性沸石,而Fe3+改性效果优于Fe2+,经FeCl3改性的沸石对菌体具有良好的吸附作用,当Fe3+-zeolite用量为180 g/L时,细胞的固定效率达到87%。在此基础上,比较了沸石负载的铁离子量对丁醇发酵性能的影响,沸石负载的铁离子量为6.0 mg/g时可显著提高丁醇发酵性能,当葡萄糖质量浓度为60 g/L时进行发酵,丁醇产量为13.5 g/L,总溶剂可达20 g/L,总溶剂的生产速率为0.385g/(L.h),比游离细胞发酵分别提高了9.5%、10.3%和40%。     

Alterations in surfaces and textures of minerals during the bacterial leaching of a complex sulfide ore

Abstract

The purpose of the work was to characterize changes in surface textures of minerals during the biological leaching of a complex sulfide ore. The ore contained pyrrhotite (Fe_I__xS), pyrite (FeS₂), sphalerite (ZnS), pentlandite [(Ni,Fe,Co)₉S₈], and chalcopyrite (CuFeS₂). Several mixed cultures were initially screened using the ore material as the sole substrate. Shake flask leaching experiments showed no major differences among test cultures, which were all derived by enrichment techniques using environmental samples collected from a mine site. Leached pyrrhotite surfaces were invariably surrounded by a dark rim of elemental S. A reaction zone was also associated with leached sphalerite grains. Chemical analyses of leach solutions indicated that the relative ranking of biological leaching of the sulfide minerals was Zn > Ni > Co > Cu. Microscopic observations were in keeping with this rankin     

资源植物罗布麻的愈伤组织诱导和植株再生研究

以罗布麻（Apocynum venetum L.）无菌苗的叶、茎和根作为外植体,在不同激素与浓度组合的MS培养基上进行愈伤组织诱导和植株再生研究。结果表明,在多个激素浓度配比的培养基中,罗布麻叶和茎的愈伤组织诱导率均可达到100%;但进一步将愈伤组织转接到含有不同激素组合的培养基进行不定芽分化时,叶和茎来源的愈伤组织的不定芽分化率有很大不同,茎来源的愈伤组织虽可在多个培养基中有不定芽分化,但最高诱导率仅达20%;而叶片来源的愈伤组织虽仅有4个激素组合培养基中有不定芽的分化,但最高分化率可达到35%。因此,构建罗布麻再生体系的最佳外植体应选择叶片。愈伤组织诱导与不定芽分化的最佳培养基均为：MS＋NAA0.1 mg/L＋6-BA 1.0 mg/L;不定芽在1/2MS＋NAA 0.2 mg/L生根培养基的生根效果最佳,不仅根群质量好,且生根率也高达100%;此再生苗的移栽成活率也最高,在适宜条件下可达75%。