Bioleaching of chalcocite by mixed microorganisms subjected to mutation

Mixed microorganisms with elevated activity of chalcocite-leaching were screened by mutation methods. The original microorganisms collected from acid mine drainage of different sites were mixed and then treated with mutagens NO₂⁻, diethyl sulfate (DES), UV and their combinations, respectively. Five groups of mixed microorganisms with much stronger ore-leaching ability were obtained by screening on the leaching media. Among them, group E of mixed microorganisms (treated with 1% DES for 60 min) with the best performance on chalcocite-leaching, increases the content of Cu²⁺ by 101.4% in 20 d of leaching compared with the control culture. In addition, group E is more tolerant to Cu²⁺ in media than the control without mutation treatment. Analysis for the diversity of microbial clones indicates that half of operational taxonomic units (OTUs) in group E are Acidithiobacillus ferrooxidans. These observations suggest that group E might have potentials for industrial application. Foundation item: Project(50321402) supported by the National Natural Science Foundation of China; Project(2004CB619201) supported by the Major State Basic Research and Development Program of China     

RAPD用于20株嗜酸氧化亚铁硫杆菌菌株亲缘性研究

为研究中国20株嗜酸氧化亚铁硫杆菌菌株的基因多样性,以及基因指纹技术分类与嗜酸氧化亚铁硫杆菌生理生化特性的相关性,使用20种随机引物,通过PCR扩增20株菌株的基因组DNA,并分析凝胶电泳条带的有无,使用Jaccard系数和UPGMA分析菌株间相关性。20种引物产生的条带将20株菌株分成了5组,菌株间相关性系数最小为35.1%(YTW与DXS),而相似系数最大为96.9%(BY-0502与DXS,DY26与DY30)。结果表明:20株来自不同地区的嗜酸氧化亚铁硫杆菌株具有较高的多样性,并且亲缘性分类与各菌株取样地以及最大铜离子耐受能力分类,有较高的分类相似性,说明RAPD技术拥有在嗜酸氧化亚铁硫杆菌内分类鉴定菌株的能力。     

两株不同铁氧化细菌合成的沉淀差异性分析

通过ICP-AES、SEM、XRD和FTIR等方法，对不同铁氧化细菌在不同温度条件下所合成的赭黄色高铁沉淀进行了分析。结果表明：无论是嗜铁钩端螺旋菌DY菌株（Leptospirillum ferriphium），还是嗜酸氧化亚铁硫杆菌GF菌株（Acidithiobacillus ferrooxidans）合成的沉淀都是黄钾铁矾和黄铵铁矾的混合物；其中，GF合成的沉淀混合物中黄钾铁矾的含量比DY合成的黄钾铁矾约高出5．53％；而DY合成的沉淀中，黄铵铁矾的含量比GF合成的沉淀中的黄铵铁矾的含量约高出15．24％。     

混合高温菌浸出黄铜矿及浸出过程中微生物群落的演替

研究3株极端嗜热古菌(金属硫叶菌,Sulfolobus metallicus JCM 9184;瑟杜生金属球菌,Metallosphaera sedula JCM 9185和万座酸菌,Acidianus manzaensis YN25)在不同起始pH值和不同温度条件下对黄铜矿的混合浸出,并对浸矿过程中混合菌群落的动态演替进行分析.结果表明:在起始pH 1.5时的铜浸出率明显高于在起始pH 2.5时的铜浸出率,而65 ℃条件下的铜浸出率高于75 ℃时的铜浸出率.利用限制性长度多态性(RFLP)分析65 ℃、起始pH 1.5条件下的微生物群落演替,结果显示:在黄铜矿的浸出前期Sulfolobus metallicus是占据优势的菌种,而到后期Acidianus manzaensis的比例则会上升,并最后取代Sulfolobus metallicus成为优势种.     

Liquid-nitrogen cryopreservation of three kinds of autotrophicbioleaching bacteria

Three kinds of autotrophic bioleaching bacteria strains, including mesophilic and acidophilic ferrous ion-oxidizing bacteria Acidithiobacillus ferrooxidans （A. ferrooxidans）, mesophilic and acidophilic sulfur-oxidizing bacteria Acidithiobacillus thiooxidans （A. thiooxidans）, and moderately thermophilic sulfur-oxidizing bacteria Acidianus brierleyi, were cryopreserved in liquid nitrogen and their ferrous ion- or sulfur-oxidizing activities were investigated and compared with the original ones. The results revealed that ferrous ion/sulfur oxidation activities of the strains were almost equal before and after cryopreservation. Glycerin was used as cryoprotective agent. In conclusion, liquid-nitrogen cryopreservation is a simple and effective method for autotrophic bioleaching microorganisms.     

Acidithiobacillus ferrooxidans菌株的分离及阴离子对其亚铁氧化活性的影响

采用连续稀释法分别从江西德兴铜矿、湖北大冶铜矿和广西大厂这3处酸性矿坑水样品中分离得到三株嗜酸性短杆状细菌,其形态及生理特征与嗜酸性氧化亚铁硫杆菌相似,16SrRNA系统学分析进一步证实这种细菌属于硫杆菌属的嗜酸性氧化亚铁硫杆菌。研究表明：阴离子对嗜酸氧化亚铁硫杆菌的Fe^2＋氧化能力具有一定影响,其影响顺序由大到小依次为：N03〉Cl^-〉SO4^2-;不同菌株对阴离子的耐受能力存在明显的差异。     

基于正交设计法的西藏玉龙铜矿酸浸工艺研究

为了提高西藏玉龙铜矿铜氧化矿的酸浸效率,采用正交设计法优化了酸浸过程中的各项参数,以期提高铜浸出效率,为中试及工程化生产提供理论依据.首先研究了氧化铜矿硫酸浸出的单因素最佳区间,然后针对温度、初始酸浓度、浸出时间、液固比和磨矿细度等多因素进行了正交优化.结果表明:这些因素对浸出过程均有影响,其中温度影响最大.通过参数优化,最终确定搅拌浸出最优工艺条件为:温度30℃、初始酸浓度100 g/L、浸矿时间2 h、液固比(指质量比)2∶1、磨矿细度为粒径≤74μm的物粒占65%.此时,铜氧化矿中铜浸出率可以达到79.45%.     

Mutagenic breeding of silver-resistant Acidithiobacillusferrooxidans and exploration of resistant mechanism

The silver-resistant Acidithiobacillus ferrooxidans were isolated from 22 acid mine drainage （AMD） samples collected from Dexing Copper Mine and Chengmen Mountain Mine, Jiangxi Province, China. Isolate DX16 is obtained from the sample taken from Dexing Copper Mine and still carries out ferrous ion oxidation when incubated in 9K medium containing silver nitrate （240 mg/L）. While isolate HI, a less resistant strain taken from Yin Mountain Mine, has a tolerate level of only 60 mg/L. Based on 16S ribosomal DNA sequencing, both bacterial 16SrDNA sequences are 100% similar to Acidithiobacillus ferrooxidans ATCC 23270. Through ultraviolet irradiation induced mutations, isolate mDX16 that is obtained from DX16 carries out ferrous ion oxidation when incubated in 9K medium containing higher concentration of silver nitrate （250 mg/L）. When silver-resistant gene （SilC） analysis is carried out on the two isolates, it is seen that this gene was absent in both.     

喜温嗜酸硫杆菌YN12菌株的鉴定及其镉抗性能

自云南酸性热泉水样中分离出一株中度嗜热硫氧化菌YN12.对其形态特征和生理生化特性以及16S rDNA序列分析结果证明,该菌株归属于喜温嗜酸硫杆菌(Acidithiobacillus caldus).重金属抗性实验表明,YN12菌株对3CdSO4-8H2O具有超强抗性,其最高初始Cd2 耐受浓度达4.8 g/L.在此基础上,不断提升3CdSO4-8H2O浓度,其最终Cd2 耐受浓度可达31.5 g /L(相当于3CdSO4-8H2O 210 g/L).在该最终Cd2 耐受浓度下,经过连续3代的适应性生长,YN12菌株的生长速度和硫氧化活性均能得到较好的恢复.