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Dissimilatory hexaheme c nitrite reductase of “Spirillum” strain 5175: purification and properties

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Abstract

When grown with nitrate as terminal electron acceptor both the soluble (periplasm, cytoplasm) and the membrane fraction of “Spirillum” strain 5175 exhibited high nitrite reductase activity. The nitrite reductase obtained from the soluble fraction was purified 76-fold to electrophoretical homogeneity. The enzyme reduced nitrite to ammonia with a specific activity of 723 μmol NO sup-inf2 × (mg protein × min)-1. The molecular mass was 58±1 kDa by SDS-PAGE compared to 59±2 kDa determined by size exclusion chromatography under nondenaturing conditions. The enzyme (as isolated) contained 5.97±0.15 heme c molecules/Mr 58 kDa. The absorption spectrum was typical for c-type cytochrome with maxima at 280, 408, 532 and 610 nm (oxidized) and at 420, 523 and 553 nm (dithionite-reduced). The enzyme (as isolated) exhibited a complex set of high-spin and lowspin ferric heme resonances with g-values at 9.82, 3,85, 3.31, 2.95, 2.30 and 1.49 in agreement with data reported for electron paramagnetic resonance spectra of nitrite reductases from Desulfovibrio desulfuricans, Wolinella succinogenes and Escherichia coli.

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Abbreviations

DNRA:

dissimilatory nitrate reduction to ammonia

EPR:

electron paramagnetic resonance

PAGE:

polyacrylamide gel electrophoresis

NaPi :

sodium phosphate

SDS:

sodium dodecylsulfate

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  1. Fakultät Biologie, Universität Konstanz, W-7750, Konstanz, Federal Republic of Germany

    Wolfram Schumacher & Peter M. H. Kroneck

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  1. Wolfram Schumacher
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  2. Peter M. H. Kroneck
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Schumacher, W., Kroneck, P.M.H. Dissimilatory hexaheme c nitrite reductase of “Spirillum” strain 5175: purification and properties. Arch. Microbiol. 156, 70–74 (1991). https://doi.org/10.1007/BF00418190

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  • DOI: https://doi.org/10.1007/BF00418190

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