Mutational Analysis of the Nor Gene Cluster which Encodes Nitric‐Oxide Reductase from Paracoccus denitrificans

作者：Anthonius P. N. De Boer, John van der Oost, Willem N. M. Reijnders, Hans Victor Westerhoff, Adriaan H. Stouthamer, Rob J. M. van Spanning · 发表于：European Journal of Biochemistry · 年份：1996 · DOI：10.1111/j.1432-1033.1996.0592r.x · 被引用次数：123 · 研究领域：Microbial Fuel Cells and Bioremediation、Photosynthetic Processes and Mechanisms、Wastewater Treatment and Nitrogen Removal

The genes that encode the hc-type nitric-oxide reductase from Paracoccus denitrificans have been identified. They are part of a cluster of six genes (norCBQDEF) and are found near the gene cluster that encodes the cd1-type nitrite reductase, which was identified earlier [de Boer, A. P. N., Reijnders, W. N. M., Kuenen, J. G., Stouthamer, A. H. & van Spanning, R. J. M. (1994) Isolation, sequencing and mutational analysis of a gene cluster involved in nitrite reduction in Paracoccus denitrificans, Antonie Leeu wenhoek 66, 111-127]. norC and norB encode the cytochrome-c-containing subunit II and cytochrome b-containing subunit I of nitric-oxide reductase (NO reductase), respectively. norQ encodes a protein with an ATP-binding motif and has high similarity to NirQ from Pseudomonas stutzeri and Pseudomonas aeruginosa and CbbQ from Pseudomonas hydrogenothermophila. norE encodes a protein with five putative transmembrane alpha-helices and has similarity to CoxIII, the third subunit of the aa3-type cytochrome-c oxidases. norF encodes a small protein with two putative transmembrane alpha-helices. Mutagenesis of norC, norB, norQ and norD resulted in cells unable to grow anaerobically. Nitrite reductase and NO reductase (with succinate or ascorbate as substrates) and nitrous oxide reductase (with succinate as substrate) activities were not detected in these mutant strains. Nitrite extrusion was detected in the medium, indicating that nitrate reductase was active. The norQ and norD mutant...