N-terminal truncation of PhaC BP-M-CPF4 and its effect on PHA production.

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  • Additional Information
    • Source:
      Publisher: BioMed Central Country of Publication: England NLM ID: 101139812 Publication Model: Electronic Cited Medium: Internet ISSN: 1475-2859 (Electronic) Linking ISSN: 14752859 NLM ISO Abbreviation: Microb Cell Fact Subsets: MEDLINE
    • Publication Information:
      Original Publication: London : BioMed Central, [2002-
    • Subject Terms:
      Polyhydroxyalkanoates*/metabolism ; Cupriavidus necator*/genetics ; Cupriavidus necator*/metabolism; 3-Hydroxybutyric Acid ; Caproates/metabolism ; Hydroxybutyrates/metabolism ; Acyltransferases/genetics ; Acyltransferases/metabolism ; Cytoplasmic Granules
    • Abstract:
      Background: Among the polyhydroxyalkanoate (PHA), poly[(R)-3-hydroxybutyrate-co-(R)-3-hydroxyhexanoate] [P(3HB-co-3HHx)] is reported to closely resemble polypropylene and low-density polyethylene. Studies have shown that PHA synthase (PhaC) from mangrove soil (PhaC BP-M-CPF4 ) is an efficient PhaC for P(3HB-co-3HHx) production and N-termini of PhaCs influence its substrate specificity, dimerization, granule morphology, and molecular weights of PHA produced. This study aims to further improve PhaC BP-M-CPF4 through N-terminal truncation.
      Results: The N-terminal truncated mutants of PhaC BP-M-CPF4 were constructed based on the information of the predicted secondary and tertiary structures using PSIPRED server and AlphaFold2 program, respectively. The N-terminal truncated PhaC BP-M-CPF4 mutants were evaluated in C. necator mutant PHB - 4 based on the cell dry weight, PHA content, 3HHx molar composition, molecular weights, and granule morphology of the PHA granules. The results showed that most transformants harbouring the N-terminal truncated PhaC BP-M-CPF4 showed a reduction in PHA content and cell dry weight except for PhaC BP-M-CPF4 G8. PhaC BP-M-CPF4 G8 and A27 showed an improved weight-average molecular weight (M w ) of PHA produced due to lower expression of the truncated PhaC BP-M-CPF4 . Transformants harbouring PhaC BP-M-CPF4 G8, A27, and T74 showed a reduction in the number of granules. PhaC BP-M-CPF4 G8 produced higher M w PHA in mostly single larger PHA granules with comparable production as the full-length PhaC BP-M-CPF4 .
      Conclusion: This research showed that N-terminal truncation had effects on PHA accumulation, substrate specificity, M w , and granule morphology. This study also showed that N-terminal truncation of the amino acids that did not adopt any secondary structure can be an alternative to improve PhaCs for the production of PHA with higher M w in mostly single larger granules.
      (© 2024. The Author(s).)
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    • Grant Information:
      203/PBIOLOGI/67811001 Ministry of Higher Education, titled "Soil Analysis and Value-Addition to Oil Palm Trunk (OPT) and sap through Biotechnology" as well as Science and Technology Research Partnership for Sustainable Development (SATREPS); 304/PBIOLOGI/6501281/F117 Zacros (Fujimori Kogyo Co. Ltd.)
    • Contributed Indexing:
      Keywords: Molecular weights; N-terminal truncation; PHA granule morphology; Poly(3-hydroxybutyrate-co-3-hydroxyhexanoate); Polyhydroxyalkanoate synthase; phaC gene expression
    • Accession Number:
      0 (Polyhydroxyalkanoates)
      TZP1275679 (3-Hydroxybutyric Acid)
      0 (Caproates)
      0 (Hydroxybutyrates)
      EC 2.3.- (Acyltransferases)
    • Publication Date:
      Date Created: 20240215 Date Completed: 20240219 Latest Revision: 20240219
    • Publication Date:
      20240219
    • Accession Number:
      PMC10867992
    • Accession Number:
      10.1186/s12934-024-02329-w
    • Accession Number:
      38360657