Yin Huaqun, Zhang Xian, Li Xiaoqi, He Zhili, Liang Yili, Guo Xue, Hu Qi, Xiao Yunhua, Cong Jing, Ma Liyuan, Niu Jiaojiao, Liu Xueduan
School of Minerals Processing and Bioengineering, Central South University, Changsha, China.
BMC Microbiol. 2014 Jul 4;14:179. doi: 10.1186/1471-2180-14-179.
Acidithiobacillus thiooxidans (A. thiooxidans), a chemolithoautotrophic extremophile, is widely used in the industrial recovery of copper (bioleaching or biomining). The organism grows and survives by autotrophically utilizing energy derived from the oxidation of elemental sulfur and reduced inorganic sulfur compounds (RISCs). However, the lack of genetic manipulation systems has restricted our exploration of its physiology. With the development of high-throughput sequencing technology, the whole genome sequence analysis of A. thiooxidans has allowed preliminary models to be built for genes/enzymes involved in key energy pathways like sulfur oxidation.
The genome of A. thiooxidans A01 was sequenced and annotated. It contains key sulfur oxidation enzymes involved in the oxidation of elemental sulfur and RISCs, such as sulfur dioxygenase (SDO), sulfide quinone reductase (SQR), thiosulfate:quinone oxidoreductase (TQO), tetrathionate hydrolase (TetH), sulfur oxidizing protein (Sox) system and their associated electron transport components. Also, the sulfur oxygenase reductase (SOR) gene was detected in the draft genome sequence of A. thiooxidans A01, and multiple sequence alignment was performed to explore the function of groups of related protein sequences. In addition, another putative pathway was found in the cytoplasm of A. thiooxidans, which catalyzes sulfite to sulfate as the final product by phosphoadenosine phosphosulfate (PAPS) reductase and adenylylsulfate (APS) kinase. This differs from its closest relative Acidithiobacillus caldus, which is performed by sulfate adenylyltransferase (SAT). Furthermore, real-time quantitative PCR analysis showed that most of sulfur oxidation genes were more strongly expressed in the S0 medium than that in the Na2S2O3 medium at the mid-log phase.
Sulfur oxidation model of A. thiooxidans A01 has been constructed based on previous studies from other sulfur oxidizing strains and its genome sequence analyses, providing insights into our understanding of its physiology and further analysis of potential functions of key sulfur oxidation genes.
嗜酸氧化硫硫杆菌(A. thiooxidans)是一种化能自养极端微生物,广泛应用于铜的工业回收(生物浸出或生物采矿)。该生物体通过自养利用元素硫和还原态无机硫化合物(RISCs)氧化产生的能量来生长和存活。然而,缺乏基因操作体系限制了我们对其生理学的探索。随着高通量测序技术的发展,嗜酸氧化硫硫杆菌的全基因组序列分析使得参与硫氧化等关键能量途径的基因/酶的初步模型得以建立。
对嗜酸氧化硫硫杆菌A01的基因组进行了测序和注释。它包含参与元素硫和RISCs氧化的关键硫氧化酶,如硫双加氧酶(SDO)、硫化物醌还原酶(SQR)、硫代硫酸盐:醌氧化还原酶(TQO)、连四硫酸盐水解酶(TetH)、硫氧化蛋白(Sox)系统及其相关的电子传递成分。此外,在嗜酸氧化硫硫杆菌A01的基因组草图序列中检测到了硫氧还蛋白还原酶(SOR)基因,并进行了多序列比对以探索相关蛋白序列组的功能。另外,在嗜酸氧化硫硫杆菌的细胞质中发现了另一条推定途径,该途径通过磷酸腺苷磷酸硫酸(PAPS)还原酶和腺苷硫酸(APS)激酶催化亚硫酸盐生成硫酸盐作为最终产物。这与其亲缘关系最近的嗜热嗜酸硫杆菌不同,后者是通过硫酸腺苷转移酶(SAT)来完成的。此外,实时定量PCR分析表明,在对数中期,大多数硫氧化基因在S0培养基中的表达比在Na2S2O3培养基中更强。
基于之前对其他硫氧化菌株的研究及其基因组序列分析,构建了嗜酸氧化硫硫杆菌A01的硫氧化模型,为我们理解其生理学以及进一步分析关键硫氧化基因的潜在功能提供了见解。