Wakeman Kathryn, Auvinen Hannele, Johnson D Barrie
School of Biological Sciences, Bangor University, Wales LL572UW, United Kingdom.
Biotechnol Bioeng. 2008 Nov 1;101(4):739-50. doi: 10.1002/bit.21951.
The evolution of microbial populations involved in simulated-heap leaching of a polymetallic black schist sulfide ore (from the recently-commissioned Talvivaara mine, Finland) was monitored in aerated packed bed column reactors over a period of 40 weeks. The influence of ore particle size (2-6.5 mm and 6.5-12 mm) on changes in composition of the bioleaching microflora and mineral leaching dynamics in columns was investigated and compared to fine-grain (<2 microm) ore that was bioprocessed in shake flask cultures. Both column reactors and shake flasks were inoculated with 24 different species and strains of mineral-oxidizing and other acidophilic micro-organisms, and maintained at 37 degrees C. Mineral oxidation was most rapid in shake flask cultures, with about 80% of both manganese and nickel and 68% of zinc being leached within 6 weeks, though relatively little of the copper present in the ore was solubilised. The microbial consortium that emerged from the original inoculum was relatively simple in shake flasks, and was dominated by the iron-oxidizing autotroph Leptospirillum ferriphilum, with smaller numbers of Acidimicrobium ferrooxidans, Acidithiobacillus caldus and Leptospirillum ferrooxidans. Both metal recovery and (for the most part) total numbers of prokaryotes were greater in the column reactor containing the medium-grain than that containing the coarse-grain ore. The bioleaching communities in the columns displayed temporal changes in composition and differed radically from those in shake flask cultures. While iron-oxidizing chemoautotrophic bacteria were always the most numerically dominant bacteria in the medium-grain column bioreactor, there were major shifts in the most abundant species present, with the type strain of Acidithiobacillus ferrooxidans dominating in the early phase of the experiment and other bacteria (At. ferrooxidans NO37 and L. ferriphilum) dominating from week 4 to week 40. With the coarse-grain column bioreactor, similar transitions in populations of iron-oxidizing chemoautotrophs were observed, though heterotrophic acidophiles were often the most abundant bacteria found in mineral leach liquors. Four bacteria not included in the mixed culture used to inoculate the columns were detected by biomolecular techniques and three of these (all Alicyclobacillus-like Firmicutes) were isolated as pure cultures. The fourth bacterium, identified from a clone library, was related to the Gram-positive sulfate reducer Desulfotomaculum salinum. All four were considered to have been present as endospores on the dried ore, which was not sterilized in the column bioreactors. Two of the Alicyclobacillus-like isolates were found, transiently, in large numbers in mineral leachates. The data support the hypothesis that temporal and spatial heterogeneity in mineral heaps create conditions that favour different mineral-oxidizing microflora, and that it is therefore important that sufficient microbial diversity is present in heaps to optimize metal extraction.
在充气填充床柱式反应器中,对参与多金属黑色片岩硫化矿(来自芬兰最近投产的塔尔维瓦拉矿)模拟堆浸的微生物种群演变进行了为期40周的监测。研究了矿石粒度(2 - 6.5毫米和6.5 - 12毫米)对柱式生物浸出微生物群落组成变化和矿物浸出动力学的影响,并与在摇瓶培养中进行生物处理的细粒(<2微米)矿石进行了比较。柱式反应器和摇瓶均接种了24种不同的矿物氧化及其他嗜酸微生物物种和菌株,并保持在37摄氏度。在摇瓶培养中矿物氧化最快,6周内约80%的锰和镍以及68%的锌被浸出,不过矿石中存在的铜相对较少被溶解。从原始接种物中出现的微生物群落,在摇瓶中相对简单,以铁氧化自养菌嗜铁钩端螺旋菌为主,嗜酸氧化亚铁微菌、嗜热嗜酸硫杆菌和嗜酸氧化亚铁钩端螺旋菌数量较少。含有中粒矿石的柱式反应器中金属回收率和(在大多数情况下)原核生物总数均高于含有粗粒矿石的柱式反应器。柱式生物浸出群落的组成随时间变化,与摇瓶培养中的群落有根本差异。虽然铁氧化化学自养菌在中粒柱式生物反应器中始终是数量上占主导的细菌,但存在的最优势物种发生了重大变化,在实验早期氧化亚铁硫杆菌的模式菌株占主导,而从第4周到第40周其他细菌(氧化亚铁硫杆菌NO37和嗜铁钩端螺旋菌)占主导。在粗粒柱式生物反应器中,观察到铁氧化化学自养菌种群有类似转变,不过异养嗜酸菌在矿物浸出液中往往是最丰富的细菌。通过生物分子技术检测到四种未包含在用于接种柱式反应器的混合培养物中的细菌,其中三种(均为类 Alicyclobacillus 的厚壁菌门细菌)被分离为纯培养物。从克隆文库鉴定出的第四种细菌与革兰氏阳性硫酸盐还原菌盐脱硫肠状菌有关。所有四种细菌被认为是以干燥矿石上的芽孢形式存在,柱式生物反应器中未对矿石进行灭菌处理。在矿物浸出液中短暂发现大量类 Alicyclobacillus 分离物中的两种。数据支持这样的假设,即矿堆中的时间和空间异质性创造了有利于不同矿物氧化微生物群落的条件,因此矿堆中存在足够的微生物多样性对于优化金属提取很重要。