Mechanism of H2S Oxidation by the Dissimilatory Perchlorate-Reducing Microorganism Azospira suillum PS

Mehta-Kolte, Misha G.; Loutey, Dana; Wang, Ouwei; Youngblut, Matthew D.; Hubbard, Christopher G.; Wetmore, Kelly M.; Conrad, Mark E.; Coates, John D.

doi:10.1128/mbio.02023-16

Mechanism of H₂S Oxidation by the Dissimilatory Perchlorate-Reducing Microorganism Azospira suillum PS

Journal Article · Mon Feb 20 23:00:00 EST 2017 · mBio (Online)

DOI:https://doi.org/10.1128/mbio.02023-16· OSTI ID:1626142

Mehta-Kolte, Misha G. ^[1]; Loutey, Dana ^[2]; Wang, Ouwei ^[3]; Youngblut, Matthew D. ^[2]; Hubbard, Christopher G. ^[4]; Wetmore, Kelly M. ^[2]; Conrad, Mark E. ^[4]; Coates, John D. ^[5]

Univ. of California, Berkeley, CA (United States). Energy Biosciences Inst.; DOE/OSTI
Univ. of California, Berkeley, CA (United States). Energy Biosciences Inst.
Univ. of California, Berkeley, CA (United States). Energy Biosciences Inst.; Univ. of California, Berkeley, CA (United States). Plant and Microbial Biology Dept.
Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States). Earth and Environmental Sciences Area
Univ. of California, Berkeley, CA (United States). Energy Biosciences Inst.; Univ. of California, Berkeley, CA (United States). Plant and Microbial Biology Dept.

The genetic and biochemical basis of perchlorate-dependent H₂S oxidation (PSOX) was investigated in the dissimilatory perchlorate-reducing microorganism (DPRM) Azospira suillum PS (PS). Previously, it was shown that all known DPRMs innately oxidize H₂S, producing elemental sulfur (S^o). Although the process involving PSOX is thermodynamically favorable (ΔG^°' = -206 kJ ∙ mol^-1 H₂S), the underlying biochemical and genetic mechanisms are currently unknown. Interestingly, H₂S is preferentially utilized over physiological electron donors such as lactate or acetate although no growth benefit is obtained from the metabolism. Here, we determined that PSOX is due to a combination of enzymatic and abiotic interactions involving reactive intermediates of perchlorate respiration. Using various approaches, including barcode analysis by sequencing (Bar-seq), transcriptome sequencing (RNA-seq), and proteomics, along with targeted mutagenesis and biochemical characterization, we identified all facets of PSOX in PS. In support of our proposed model, deletion of identified upregulated PS genes traditionally known to be involved in sulfur redox cycling (e.g., Sox, sulfide:quinone reductase [SQR]) showed no defect in PSOX activity. Proteomic analysis revealed differential abundances of a variety of stress response metal efflux pumps and divalent heavy-metal transporter proteins, suggesting a general toxicity response. Furthermore, in vitro biochemical studies demonstrated direct PSOX mediated by purified perchlorate reductase (PcrAB) in the absence of other electron transfer proteins. The results of these studies support a model in which H₂S oxidation is mediated by electron transport chain short-circuiting in the periplasmic space where the PcrAB directly oxidizes H₂S to S^o. The biogenically formed reactive intermediates (ClO₂^- and O₂) subsequently react with additional H₂S, producing polysulfide and S^o as end products.

View Accepted Manuscript (DOE)

Research Organization:: Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States)

Sponsoring Organization:: USDOE Office of Science (SC)

Grant/Contract Number:: AC02-05CH11231

OSTI ID:: 1626142

Journal Information:: mBio (Online), Journal Name: mBio (Online) Journal Issue: 1 Vol. 8; ISSN 2150-7511

Publisher:: American Society for Microbiology (ASM)Copyright Statement

Country of Publication:: United States

Language:: English

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