Schmidt Astrid, Schmidt André, Haferburg Götz, Kothe Erika
Microbial Phytopathology, Institute of Microbiology, Friedrich-Schiller-University, Jena, Germany.
J Basic Microbiol. 2007 Feb;47(1):56-62. doi: 10.1002/jobm.200610213.
Heavy metal tolerant and resistant strains of streptomycetes isolated from a former uranium mining site were screened for their superoxide dismutase expression. From the strains tolerating high concentrations of different heavy metals, one was selected for its tolerance of concentrations of heavy metals (Ni, Cu, Cd, Cr, Mn, Zn, Fe). This strain, Streptomyces acidiscabies E13, was chosen for the purpose of superoxide dismutase analysis. Gel electrophoresis and activity staining revealed only one each of a nickel (NiSOD) and an iron (FeZnSOD) containing superoxide dismutase as shown by differential enzymatic repression studies. The gene for nickel containing superoxide dismutase, sodN, was cloned and sequenced from this strain. The genomic sequence shows 92.7% nucleotide identity and 96.1% amino acid identity to sodN of S. coelicolor. Expression can be activated by nickel as well as other heavy metals and active enzyme is produced in media lacking nickel but containing copper, iron or zinc. Thus, the selected strain is well suited for further characterization of the enzyme encoded by sodN.
对从一个以前的铀矿开采场地分离出的耐重金属和抗重金属链霉菌菌株进行了超氧化物歧化酶表达筛选。从耐受不同高浓度重金属的菌株中,选择了一株对多种重金属(镍、铜、镉、铬、锰、锌、铁)具有耐受性的菌株。该菌株,即疮痂链霉菌E13,被选用于超氧化物歧化酶分析。凝胶电泳和活性染色显示,如差异酶抑制研究所表明的那样,仅存在一种含镍超氧化物歧化酶(NiSOD)和一种含铁超氧化物歧化酶(FeZnSOD)。从该菌株中克隆并测序了含镍超氧化物歧化酶的基因sodN。基因组序列与天蓝色链霉菌的sodN显示出92.7%的核苷酸同一性和96.1%的氨基酸同一性。镍以及其他重金属均可激活该基因的表达,并且在不含镍但含有铜、铁或锌的培养基中可产生活性酶。因此,所选菌株非常适合对sodN编码的酶进行进一步表征。