acI Actinobacteria Assemble a Functional Actinorhodopsin with Natively Synthesized Retinal
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acI Actinobacteria Assemble a Functional Actinorhodopsin with Natively Synthesized Retinal
  • Published Date:

    12/15/2018

  • Source:
    Appl Environ Microbiol. 2018 Dec 15; 84(24): e01678-18.
Filetype[PDF-1.19 MB]


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  • Description:
    Freshwater lakes harbor complex microbial communities, but these ecosystems are often dominated by acI Actinobacteria. Members of this cosmopolitan lineage are proposed to bolster heterotrophic growth using phototrophy because their genomes encode actino-opsins (actR). This model has been difficult to validate experimentally because acI Actinobacteria are not consistently culturable. Based primarily on genomes from single cells and metagenomes, we provide a detailed biosynthetic route for members of acI clades A and B to synthesize retinal and its carotenoid precursors. Consequently, acI cells should be able to natively assemble light-driven actinorhodopsins (holo-ActR) to pump protons, unlike many bacteria that encode opsins but may need to exogenously obtain retinal because they lack retinal machinery. Moreover, we show that all acI clades contain genes for a secondary branch of the carotenoid pathway, implying synthesis of a complex carotenoid. Transcription analysis of acI Actinobacteria in a eutrophic lake shows that all retinal and carotenoid pathway operons are transcribed and that actR is among the most highly transcribed of all acI genes. Furthermore, heterologous expression of acI retinal pathway genes showed that lycopene, retinal, and ActR can be made using the genes encoded in these organisms. Model cells producing ActR and the key acI retinal-producing β-carotene oxygenase formed holo-ActR and acidified solution during illumination. Taken together, our results prove that acI Actinobacteria containing both ActR and acI retinal production machinery have the capacity to natively synthesize a green light-dependent outward proton-pumping rhodopsin.
  • Pubmed ID:
    30315080
  • Pubmed Central ID:
    PMC6275354
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