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Literature summary extracted from

  • Du, J.; Rehm, B.
    Purification of target proteins from intracellular inclusions mediated by intein cleavable polyhydroxyalkanoate synthase fusions (2017), Microb. Cell Fact., 16, 184 .
    View publication on PubMedView publication on EuropePMC

Application

EC Number Application Comment Organism
2.3.1.304 biotechnology recombinant protein production and purification from Escherichia coli is often accompanied with expensive and complicated procedures, especially for therapeutic proteins. By using an intein cleavable polyhydroxyalkanoate synthase fusion, recombinant proteins can be first produced and sequestered on a natural resin, the polyhydroxyalkanoate (PHA) inclusions, then separated from contaminating host proteins via simple PHA bead isolation steps, and finally purified by specific release into the soluble fraction induced by a pH reduction Cupriavidus necator

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.3.1.304 recombinant expression of the enzyme used as a fusion protein with a self-cleaving intein linker for production of the fused proteins, e.g. Aequorea victoria green fluorescent protein (GFP), Mycobacterium tuberculosis vaccine candidate Rv1626, the immunoglobulin G (IgG) binding ZZ domain of protein A derived from Staphylococcus aureus, human tumor necrosis factor alpha (TNFalpha), human granulocyte colony-stimulating factor (G-CSF), and human interferon alpha 2beta (IFNalpha2beta) in the Escherichia coli system. A pH or thiol inducible intein is employed in combination with PHA beads for target protein production and purification, method evaluation, overview Cupriavidus necator

Protein Variants

EC Number Protein Variants Comment Organism
2.3.1.304 additional information translationally fusing a target protein to PHA synthase using a self-cleaving intein as linker, intracellular production of PHA beads is achieved. Upon isolation of respective PHA beads the soluble pure target protein is released by a simple pH shift to pH 6.0. The utility of this approach is exemplified by producing six target proteins, including Aequorea victoria green fluorescent protein (GFP), Mycobacterium tuberculosis vaccine candidate Rv1626, the immunoglobulin G (IgG) binding ZZ domain of protein A derived from Staphylococcus aureus, human tumor necrosis factor alpha (TNFalpha), human granulocyte colony-stimulating factor (G-CSF), and human interferon alpha 2beta (IFNalpha2beta) Cupriavidus necator

Organism

EC Number Organism UniProt Comment Textmining
2.3.1.304 Cupriavidus necator P23608 i.e. Ralstonia eutropha
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2.3.1.304 Cupriavidus necator ATCC 17699 P23608 i.e. Ralstonia eutropha
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2.3.1.304 Cupriavidus necator DSM 428 P23608 i.e. Ralstonia eutropha
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2.3.1.304 Cupriavidus necator Stanier 337 P23608 i.e. Ralstonia eutropha
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Synonyms

EC Number Synonyms Comment Organism
2.3.1.304 PHA synthase
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 Cupriavidus necator
2.3.1.304 polyhydroxyalkanoate synthase
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 Cupriavidus necator