Engineering of the Long-Main-Chain Monomer-Incorporating Polyhydroxyalkanoate Synthase PhaC AR for the Biosynthesis of Poly[( R )-3-hydroxybutyrate- co -6-hydroxyhexanoate].

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  • Additional Information
    • Source:
      Publisher: American Chemical Society Country of Publication: United States NLM ID: 100892849 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1526-4602 (Electronic) Linking ISSN: 15257797 NLM ISO Abbreviation: Biomacromolecules Subsets: MEDLINE
    • Publication Information:
      Original Publication: Washington, DC : American Chemical Society, c2000-
    • Subject Terms:
      Acyltransferases*/genetics ; Acyltransferases*/metabolism ; Escherichia coli*/genetics ; Escherichia coli*/metabolism ; Caproates*/chemistry ; Caproates*/metabolism; Protein Engineering/methods ; Polyesters/chemistry ; Polyesters/metabolism ; Mutagenesis, Site-Directed ; Polyhydroxyalkanoates/chemistry ; Polyhydroxyalkanoates/biosynthesis ; Bacterial Proteins/genetics ; Bacterial Proteins/metabolism ; Bacterial Proteins/chemistry
    • Abstract:
      Polyhydroxyalkanoate (PHA) synthases (PhaCs) are useful and versatile tools for the production of aliphatic polyesters. Here, the chimeric PHA synthase PhaC AR was engineered to increase its capacity to incorporate unusual 6-hydroxyhexanoate (6HHx) units. Mutations at positions 149 and 314 in PhaC AR were previously found to increase the incorporation of an analogous natural monomer, 3-hydroxyhexanoate (3HHx). We attempted to repurpose the mutations to produce 6HHx-containing polymers. Site-directed saturation mutants at these positions were applied for P(3HB- co -6HHx) synthesis in Escherichia coli . As a result, the N149D and F314Y mutants effectively increased the 6HHx fraction. Moreover, the pairwise NDFY mutation further increased the 6HHx fraction, which reached 22 mol %. This increase was presumably caused by altered enzyme activity rather than altered expression levels, as assessed based on immunoblot analysis. The glass transition temperature and crystallinity of P(3HB- co -6HHx) decreased as the 6HHx fraction increased.
    • Accession Number:
      EC 2.3.- (Acyltransferases)
      EC 2.3.1.- (poly(3-hydroxyalkanoic acid) synthase)
      0 (Caproates)
      0 (Polyesters)
      0 (Polyhydroxyalkanoates)
      0 (Bacterial Proteins)
    • Publication Date:
      Date Created: 20240408 Date Completed: 20240513 Latest Revision: 20241030
    • Publication Date:
      20241031
    • Accession Number:
      10.1021/acs.biomac.4c00116
    • Accession Number:
      38588330