一种浸矿混合菌种的筛选、鉴定及浸矿的研究

doi:10.13560/j.cnki.biotech.bull.1985.2016.08.026

摘要/Abstract

摘要： 旨在从某低品位硫化铜矿酸性矿坑水中分离获得浸矿效果优良的细菌,同时对细菌的浸矿行为进行初步的研究。采集福建某低品位硫化铜矿酸性矿坑水样品,9K培养基富集获得混合菌种FIM-201304。利用9K固体平板,从混合菌FIM-201304中分离获得优势浸矿菌D-1。应用高通量Illumina Miseq测序技术分析混合菌FIM-201304 的群落结构。通过表型特征和16S rRNA基因序列分析鉴定菌株D-1。采用摇瓶培养对比研究混合菌和单菌对低品位硫化铜矿和低品位硫化镍矿的浸出效果。结果显示,高通量测序技术分析结果表明,混合菌FIM-201304 的优势菌是嗜酸氧化亚铁硫杆菌（Acidithiobacillus ferrooxidans,A. f errooxidans）（丰度占比85.02%）、铜绿色假单胞菌（Pseudomonas sp.）（丰度占比10.07%）、嗜酸异养菌（Acidiphilium acidophilum,A.acidophilum）（丰度占比1.30%）和鞘氨醇单胞菌（Sphingomonas sp.）（丰度占比1.21%）。表型特征和16S rRNA 基因序列分析确定菌株D-1属于氧化亚铁硫杆菌。摇瓶实验结果表明,浸出反应20 d后,接种混合菌的浸出体系中铜、镍浸出率分别为65%和56%,相同条件下,接种单菌的浸出体系中铜、镍的浸出率为41%和36%,接种混合菌的浸出体系比接种单菌的浸出体系中铜、镍浸出率分别提高了24%和20%。混合菌对矿石的浸出效果显著优于单菌。异养菌促进了自养菌对矿石中金属元素的浸出。

关键词: 嗜酸氧化亚铁硫杆菌, 生物浸出, 混合菌, 高通量测序, 16S rRNA 基因

Abstract: It is aimed to screen the highly efficient leaching bacteria from acid mining water of low-grade copper sulfide mine,and also preliminarily investigate the bioleaching capacities of the bacteria. Firstly,a mixed culture FIM-201304 was obtained by 9K enrichment from acid mining water of a low-grade copper sulfide mine in Fujian province. Then,one dominated bacterium D-1 was isolated from the FIM-201304 using 9K solid culture medium. Sequencing technology by high-throughput Illumina Miseq was used to analyze the community structure of FIM-201304. D-1 was identified by phenotypic characteristics and 16S rRNA gene sequencing analysis. Flask experiment was utilized to compare the effects of leaching low-grade copper- and nickel-sulfide between the mixed culture FIM-201304 and a single bacterium D-1. Results from high-throughput sequencing analysis showed that the mixed culture FIM-201304 contained the dominated bacteria of Acidithiobacillus ferrooxidans（Abundance accounted for 85.02%）,Pseudomonas sp.（Abundance accounted for 10.07%）,Acidiphilium acidophilum（Abundance accounted for 1.30%）and Sphingomonas sp.（Abundance accounted for 1.21%）. According to phenotypic characteristics and 16S rRNA gene sequence analysis,strain D-1 was identified as Acidithiobacillus ferrooxidans.The results after 20 days of bioleaching with flasks showed that leaching ratio of copper and nickel by FIM-201304 was 65% and 56% respectively,while the leaching ratio by D-1 was 41% and 36% under the same conditions,i.e.,the bioleaching ratio of copper and nickel by mixed culture increased 24% and 20% compared to that by single bacterium,respectively. Conclusively,the mixed culture resulted in the better leaching effect as heterotrophic bacteria promoted the metal-leaching capacity of autotrophic bacteria from the ore.

Key words: Acidithiobacillus ferrooxidans, bioleaching, mixed culture, high-throughput sequencing, 16S rRNA gene

聂毅磊,陈宏,罗立津,贾纬,陈星伟. 一种浸矿混合菌种的筛选、鉴定及浸矿的研究[J]. 生物技术通报, 2016, 32(8): 177-183.

NIE Yi-lei ,CHEN Hong ,LUO Li-jin, JIA Wei ,CHEN Xing-wei. Screening and Identification of Mixed Culture,and Its Bioleaching Capacity[J]. Biotechnology Bulletin, 2016, 32(8): 177-183.

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