Cloning and heterologous expression of a sulfur oxygenase/reductase gene from the thermoacidophilic archaeon Acidianus sp. S5 in Escherichia coli.
A thermoacidophilic, obligately chemolithotrophic, facultatively aerobic archaebacterium, Acidianus sp. S5, was isolated from acidothermal springs in southwest China. The sulfur oxygenase/reductase (SOR) gene of Acidianus sp. S5 was cloned and expressed in Escherichia coli. Several primers were designed and successfully applied for detection and cloning of the sor gene. A 3.7-kb EcoRI fragment containing the sor gene and three neighboring open reading frames was sequenced. Sequence analysis indicated that the sor gene of Acidianus sp. S5 showed 81% identity to the sor gene of Acidianus ambivalens. E. coli cells carrying the sor gene on pBV220SOR were able to overproduce SOR upon a temperature shift from 30 to 42 degrees C. SOR produced in E. coli catalyzes the oxidation of elemental sulfur and concomitant production of sulfite, thiosulfate and hydrogen sulfide. The recombinant enzyme exhibits the same catalytic properties as the one from Acidianus S5.
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State Key Laboratory of Institute of Microbiology, Chinese Academy of Sciences, 100080, Beijing, PR China.
Li
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Y
Zhou
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Liu
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S
D000595:Amino Acid Sequence; D019843:Archaeal Proteins; D003001:Cloning, Molecular; D004275:DNA, Ribosomal / Q000032:analysis; D015246:Deoxyribonuclease EcoRI / Q000378:metabolism; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics*; D015870:Gene Expression; D006863:Hydrogen-Ion Concentration; D008969:Molecular Sequence Data; D010084:Oxidation-Reduction; D010088:Oxidoreductases / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics*; D050862:Oxidoreductases Acting on Sulfur Group Donors; D010802:Phylogeny; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis; D017422:Sequence Analysis, DNA; D016943:Sulfolobaceae / Q000201:enzymology* / Q000235:genetics / Q000302:isolation & purification; D013455:Sulfur / Q000378:metabolism; D013696:Temperature
D019843:Archaeal Proteins; D004275:DNA, Ribosomal; D012336:RNA, Ribosomal, 16S; D011994:Recombinant Proteins; D013455:Sulfur; D010088:Oxidoreductases; D050862:Oxidoreductases Acting on Sulfur Group Donors; C058771:soR protein, Acidianus ambivalens; D015246:Deoxyribonuclease EcoRI