Cyanobacterial Sfp-type phosphopantetheinyl transferases functionalize carrier proteins of diverse biosynthetic pathways.

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  • Additional Information
    • Source:
      Publisher: Nature Publishing Group Country of Publication: England NLM ID: 101563288 Publication Model: Electronic Cited Medium: Internet ISSN: 2045-2322 (Electronic) Linking ISSN: 20452322 NLM ISO Abbreviation: Sci Rep Subsets: MEDLINE
    • Publication Information:
      Original Publication: London : Nature Publishing Group, copyright 2011-
    • Subject Terms:
      Bacterial Proteins*/chemistry ; Bacterial Proteins*/genetics ; Bacterial Proteins*/metabolism ; Biosynthetic Pathways* ; Carrier Proteins*/chemistry ; Carrier Proteins*/genetics ; Carrier Proteins*/metabolism ; Synechocystis*/enzymology ; Synechocystis*/genetics ; Transferases (Other Substituted Phosphate Groups)*/chemistry ; Transferases (Other Substituted Phosphate Groups)*/genetics ; Transferases (Other Substituted Phosphate Groups)*/metabolism; Bacillus subtilis/enzymology ; Bacillus subtilis/genetics ; Polyketides/chemistry ; Polyketides/metabolism ; Streptomyces/enzymology ; Streptomyces/genetics
    • Abstract:
      Cyanobacteria produce structurally and functionally diverse polyketides, nonribosomal peptides and their hybrids. Sfp-type phosphopantetheinyl transferases (PPTases) are essential to the production of these compounds via functionalizing carrier proteins (CPs) of biosynthetic megaenzymes. However, cyanobacterial Sfp-type PPTases remain poorly characterized, posing a significant barrier to the exploitation of cyanobacteria for biotechnological and biomedical applications. Herein, we describe the detailed characterization of multiple cyanobacterial Sfp-type PPTases that were rationally selected. Biochemical characterization of these enzymes along with the prototypic enzyme Sfp from Bacillus subtilis demonstrated their varying specificities toward 11 recombinant CPs of different types of biosynthetic pathways from cyanobacterial and Streptomyces strains. Kinetic analysis further indicated that PPTases possess the higher binding affinity and catalytic efficiency toward their cognate CPs in comparison with noncognate substrates. Moreover, when chromosomally replacing the native PPTase gene of Synechocystis sp. PCC6803, two selected cyanobacterial PPTases and Sfp supported the growth of resulted mutants. Cell lysates of the cyanobacterial mutants further functionalized recombinant CP substrates. Collectively, these studies reveal the versatile catalysis of selected cyanobacterial PPTases and provide new tools to synthesize cyanobacterial natural products using in vitro and in vivo synthetic biology approaches.
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    • Accession Number:
      0 (Bacterial Proteins)
      0 (Carrier Proteins)
      0 (Polyketides)
      0 (phosphopantetheinyl transferase)
      EC 2.7.8.- (Transferases (Other Substituted Phosphate Groups))
    • Publication Date:
      Date Created: 20170921 Date Completed: 20190625 Latest Revision: 20190625
    • Publication Date:
      20231215
    • Accession Number:
      PMC5605751
    • Accession Number:
      10.1038/s41598-017-12244-3
    • Accession Number:
      28928426