Dataset Information

Draft Whole-Genome Sequence of the Green Sulfur Photosynthetic Bacterium Chlorobaculum sp. Strain 24CR, Isolated from the Carmel River.

ABSTRACT: Green sulfur bacteria are in the family Chlorobiaceae, which is composed of four distinct genera, namely, Chlorobaculum, Chlorobium, Prosthecochloris, and Chloroherpeton, with Chlorobium species being the most commonly represented in genome studies. We have now sequenced only the fourth species of Chlorobaculum, which established Chlorobaculum sp. 24CR as a separate species and should help characterize the genus.

SUBMITTER: Freed S

PROVIDER: S-EPMC6430317 | biostudies-literature | 2019 Mar

REPOSITORIES: biostudies-literature

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Draft Whole-Genome Sequence of the Green Sulfur Photosynthetic Bacterium Chlorobaculum sp. Strain 24CR, Isolated from the Carmel River.

Freed Shawn S Robertson Sydney S Meyer Terry T Kyndt John J

Microbiology resource announcements 20190321 12

Green sulfur bacteria are in the family Chlorobiaceae, which is composed of four distinct genera, namely, Chlorobaculum, Chlorobium, Prosthecochloris, and Chloroherpeton, with Chlorobium species being the most commonly represented in genome studies. We have now sequenced only the fourth species of Chlorobaculum, which established Chlorobaculum sp. 24CR as a separate species and should help characterize the genus. ...[more]

PMID: 30938700

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Project description:Malonyl-CoA reductase (MCR) is a NADPH-dependent bi-functional enzyme that performs alcohol dehydrogenase and aldehyde dehydrogenase (CoA-acylating) activities in the N- and C-terminal fragments, respectively. It catalyzes the two-step reduction of malonyl-CoA to 3-hydroxypropionate (3-HP), a key reaction in the autotrophic CO2 fixation cycles of Chloroflexaceae green non-sulfur bacteria and the archaea Crenarchaeota. However, the structural basis underlying substrate selection, coordination, and the subsequent catalytic reactions of full-length MCR is largely unknown. For the first time, we here determined the structure of full-length MCR from the photosynthetic green non-sulfur bacterium Roseiflexus castenholzii (RfxMCR) at 3.35 Å resolution. Furthermore, we determined the crystal structures of the N- and C-terminal fragments bound with reaction intermediates NADP+ and malonate semialdehyde (MSA) at 2.0 Å and 2.3 Å, respectively, and elucidated the catalytic mechanisms using a combination of molecular dynamics simulations and enzymatic analyses. Full-length RfxMCR was a homodimer of two cross-interlocked subunits, each containing four tandemly arranged short-chain dehydrogenase/reductase (SDR) domains. Only the catalytic domains SDR1 and SDR3 incorporated additional secondary structures that changed with NADP+-MSA binding. The substrate, malonyl-CoA, was immobilized in the substrate-binding pocket of SDR3 through coordination with Arg1164 and Arg799 of SDR4 and the extra domain, respectively. Malonyl-CoA was successively reduced through protonation by the Tyr743-Arg746 pair in SDR3 and the catalytic triad (Thr165-Tyr178-Lys182) in SDR1 after nucleophilic attack from NADPH hydrides. IMPORTANCE The bi-functional MCR catalyzes NADPH-dependent reduction of malonyl-CoA to 3-HP, an important metabolic intermediate and platform chemical, from biomass. The individual MCR-N and MCR-C fragments, which contain the alcohol dehydrogenase and aldehyde dehydrogenase (CoA-acylating) activities, respectively, have previously been structurally investigated and reconstructed into a malonyl-CoA pathway for the biosynthetic production of 3-HP. However, no structural information for full-length MCR has been available to illustrate the catalytic mechanism of this enzyme, which greatly limits our capacity to increase the 3-HP yield of recombinant strains. Here, we report the cryo-electron microscopy structure of full-length MCR for the first time and elucidate the mechanisms underlying substrate selection, coordination, and catalysis in the bi-functional MCR. These findings provide a structural and mechanistic basis for enzyme engineering and biosynthetic applications of the 3-HP carbon fixation pathways.

Dataset Information

Draft Whole-Genome Sequence of the Green Sulfur Photosynthetic Bacterium Chlorobaculum sp. Strain 24CR, Isolated from the Carmel River.

Publications

Draft Whole-Genome Sequence of the Green Sulfur Photosynthetic Bacterium <i>Chlorobaculum</i> sp. Strain 24CR, Isolated from the Carmel River.

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