Characterization of the PduS cobalamin reductase of Salmonella enterica and its role in the Pdu microcompartment.

Journal of Bacteriology
Shouqiang Cheng, Thomas A Bobik

Abstract

Salmonella enterica degrades 1,2-propanediol (1,2-PD) in a coenzyme B12 (adenosylcobalamin, AdoCbl)-dependent fashion. Salmonella obtains AdoCbl by assimilation of complex precursors, such as vitamin B12 and hydroxocobalamin. Assimilation of these compounds requires reduction of their central cobalt atom from Co3+ to Co2+ to Co+, followed by adenosylation to AdoCbl. In this work, the His6-tagged PduS cobalamin reductase from S. enterica was produced at high levels in Escherichia coli, purified, and characterized. The anaerobically purified enzyme reduced cob(III)alamin to cob(II)alamin at a rate of 42.3±3.2 μmol min(-1) mg(-1), and it reduced cob(II)alamin to cob(I)alamin at a rate of 54.5±4.2 nmol min(-1) mg(-1) protein. The apparent Km values of PduS-His6 were 10.1±0.7 μM for NADH and 67.5±8.2 μM for hydroxocobalamin in cob(III)alamin reduction. The apparent Km values for cob(II)alamin reduction were 27.5±2.4 μM with NADH as the substrate and 72.4±9.5 μM with cob(II)alamin as the substrate. High-performance liquid chromatography (HPLC) and mass spectrometry (MS) indicated that each monomer of PduS contained one molecule of noncovalently bound flavin mononucleotide (FMN). Genetic studies showed that a pduS deletion decreased t...Continue Reading

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Citations

Sep 28, 2010·The Journal of Biological Chemistry·Christopher S CrowleyTodd O Yeates
Apr 16, 2013·Journal of Bacteriology·Douglas L Huseby, John R Roth
May 19, 2012·Nature Chemical Biology·Christina M AgapakisPamela A Silver
Jan 1, 2013·Current Opinion in Biotechnology·Edward Y Kim, Danielle Tullman-Ercek
Dec 3, 2014·Trends in Microbiology·Cheryl A Kerfeld, Onur Erbilgin
Sep 18, 2012·Journal of Biotechnology·Stefanie FrankMartin J Warren
Jun 10, 2014·Bioscience Reports·Andrew D LawrenceMartin J Warren
Aug 8, 2014·Journal of Molecular Biology·Michael C ThompsonTodd O Yeates
Oct 24, 2014·PLoS Computational Biology·Seth D AxenCheryl A Kerfeld
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Feb 18, 2014·Journal of Bacteriology·Elitza I TochevaGrant J Jensen
Sep 4, 2014·Microbiology and Molecular Biology Reviews : MMBR·Chiranjit ChowdhuryThomas A Bobik
Jun 27, 2019·Biochemical Society Transactions·Sara Planamente, Stefanie Frank
Apr 26, 2020·Nature Communications·Mengru YangLu-Ning Liu
Oct 9, 2020·EcoSal Plus·Katie L StewartThomas A Bobik

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