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Literature summary extracted from

  • Sznajder, A.; Jendrossek, D.
    To be or not to be a poly(3-hydroxybutyrate) (PHB) depolymerase: PhaZd1 (PhaZ6) and PhaZd2 (PhaZ7) of Ralstonia eutropha, highly active PHB depolymerases with no detectable role in mobilization of accumulated PHB (2014), Appl. Environ. Microbiol., 80, 4936-4946.
    View publication on PubMedView publication on EuropePMC

Activating Compound

EC Number Activating Compound Comment Organism Structure
3.1.1.75 EDTA activates alone, but inhibits in presence of divalent cations Cupriavidus necator

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.1.1.75 gene H16_B2073, promoter determination, subcloning and expression in Escherichia coli strains JM109 and S17-1, expression of inactive yellow fluorescent protein-tagged mutant S190A, colocalization of the PhaZd1 fusions with PHB granules, expression of N-terminally His6-tagged wild-type and mutant enzymes in Escherichia coli Cupriavidus necator
3.1.1.75 gene H16_B2401, promoter determination, subcloning and expression in Escherichia coli strains JM109 and S17-1, expression of inactive C-terminally yellow fluorescent protein-tagged mutant S193A, no colocalization with PHB granules, expression of N-terminally His6-tagged wild-type and mutant enzymes in Escherichia coli Cupriavidus necator

Protein Variants

EC Number Protein Variants Comment Organism
3.1.1.75 additional information deletion of phaZd1 without effect on enzyme activity Cupriavidus necator
3.1.1.75 additional information deletion of phaZd2 without effect on enzyme activity Cupriavidus necator
3.1.1.75 S190A site-directed mutagenesis, inactive mutant Cupriavidus necator
3.1.1.75 S193A site-directed mutagenesis, inactive mutant Cupriavidus necator

Inhibitors

EC Number Inhibitors Comment Organism Structure
3.1.1.75 Ca2+ activates up to 10 mM, inhibits above; activates up to 10 mM, inhibits above Cupriavidus necator
3.1.1.75 DTE
-
 Cupriavidus necator
3.1.1.75 EDTA activates alone, but inhibits in presence of divalent cations; activates alone, but inhibits in presence of divalent cations Cupriavidus necator
3.1.1.75 iodoacetamide
-
 Cupriavidus necator
3.1.1.75 Mg2+ activates up to 10 mM, inhibits above; activates up to 10 mM, inhibits above Cupriavidus necator
3.1.1.75 additional information activation of PhaZd2 depolymerase activity by salts, but salts affect the activity of PhaZd2 with nPHB granules as substrate but have no significant influence on the hydrolysis of p-nitrophenyl esters such as p-nitrophenyl butyrate. Poor effect by DTT; activation of PhaZd2 depolymerase activity by salts, but salts affect the activity of PhaZd2 with nPHB granules as substrate but have no significant influence on the hydrolysis of p-nitrophenyl esters such as p-nitrophenylbutyrate. Poor effect by DTT Cupriavidus necator
3.1.1.75 PMSF
-
 Cupriavidus necator
3.1.1.75 SDS almost complete inhibition at 0.01% v/v; almost complete inhibition at 0.01% v/v Cupriavidus necator
3.1.1.75 Triton X-100 almost complete inhibition at 0.01% v/v; almost complete inhibition at 0.01% v/v Cupriavidus necator

Localization

EC Number Localization Comment Organism GeneOntology No. Textmining
3.1.1.75 intracellular
-
 Cupriavidus necator 5622
-
3.1.1.75 additional information the enzyme is not detected in the PHB granule fraction Cupriavidus necator
-
-

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
3.1.1.75 Ca2+ activates up to 10 mM, inhibits above Cupriavidus necator
3.1.1.75 K+ activates Cupriavidus necator
3.1.1.75 Mg2+ activates up to 10 mM, inhibits above Cupriavidus necator
3.1.1.75 additional information activation of PhaZd2 depolymerase activity by salts, but salts affect the activity of PhaZd2 with nPHB granules as substrate but have no significant influence on the hydrolysis of p-nitrophenyl esters such as p-nitrophenylbutyrate Cupriavidus necator
3.1.1.75 additional information slight activation of PhaZd2 depolymerase activity by salts, but salts affect the activity of PhaZd1 with nPHB granules as substrate but have no significant influence on the hydrolysis of p-nitrophenyl esters such as p-nitrophenylbutyrate Cupriavidus necator

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
3.1.1.75 38400
-
 x * 38400, about, sequence calculation Cupriavidus necator
3.1.1.75 39200
-
 x * 39200, about, sequence calculation Cupriavidus necator

Organism

EC Number Organism UniProt Comment Textmining
3.1.1.75 Cupriavidus necator Q0JYJ1 gene H16_B2401
-
3.1.1.75 Cupriavidus necator Q0JZG9 gene H16_B2073
-
3.1.1.75 Cupriavidus necator H16 / ATCC 23440 / NCIB 10442 / S-10-1 Q0JYJ1 gene H16_B2401
-
3.1.1.75 Cupriavidus necator H16 / ATCC 23440 / NCIB 10442 / S-10-1 Q0JZG9 gene H16_B2073
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.1.1.75 recombinant N-terminally His6-tagged wild-type and mutant enzymes from Escherichia coli by nicke affinity chromatography Cupriavidus necator

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.1.1.75 4-nitrophenyl butyrate + H2O
-
 Cupriavidus necator 4-nitrophenol + butyrate
-
 ?
3.1.1.75 4-nitrophenyl butyrate + H2O
-
 Cupriavidus necator H16 / ATCC 23440 / NCIB 10442 / S-10-1 4-nitrophenol + butyrate
-
 ?
3.1.1.75 poly(3-hydroxybutyrate) + H2O
-
 Cupriavidus necator ?
-
 ?
3.1.1.75 poly(3-hydroxybutyrate) + H2O
-
 Cupriavidus necator H16 / ATCC 23440 / NCIB 10442 / S-10-1 ?
-
 ?

Subunits

EC Number Subunits Comment Organism
3.1.1.75 ? x * 38400, about, sequence calculation Cupriavidus necator
3.1.1.75 ? x * 39200, about, sequence calculation Cupriavidus necator

Synonyms

EC Number Synonyms Comment Organism
3.1.1.75 PhaZ6
-
 Cupriavidus necator
3.1.1.75 PhaZ7
-
 Cupriavidus necator
3.1.1.75 PhaZd1
-
 Cupriavidus necator
3.1.1.75 PhaZd2
-
 Cupriavidus necator
3.1.1.75 PHB depolymerase
-
 Cupriavidus necator
3.1.1.75 poly(3-hydroxybutyrate) depolymerase
-
 Cupriavidus necator

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.1.1.75 8.5
-
 assay at Cupriavidus necator

General Information

EC Number General Information Comment Organism
3.1.1.75 malfunction chromosomal deletion of phaZd1, phaZd2, or both depolymerase genes has no significant effect on PHB accumulation and mobilization during growth in nutrient broth or nutrient broth-gluconate medium Cupriavidus necator
3.1.1.75 additional information active site Ser190 Cupriavidus necator
3.1.1.75 additional information active site Ser193 Cupriavidus necator
3.1.1.75 physiological function PhaZd1 and is a poly(3-hydroxybutyrate) depolymerase with a high capacity to degrade poly(3-hydroxybutyrate) when artificially expressed but is apparently not involved in poly(3-hydroxybutyrate) mobilization in the wild-type. But constitutive expression of PhaZd1 reduces or even prevents the accumulation of poly(3-hydroxybutyrate) under poly(3-hydroxybutyrate)-permissive conditions in vivo Cupriavidus necator
3.1.1.75 physiological function PhaZd1 and is a poly(3-hydroxybutyrate) depolymerase with a high capacity to degrade poly(3-hydroxybutyrate) when artificially expressed but is apparently not involved in poly(3-hydroxybutyrate) mobilization in the wild-type. But constitutive expression of PhaZd2 reduces or even prevents the accumulation of poly(3-hydroxybutyrate) under poly(3-hydroxybutyrate)-permissive conditions in vivo Cupriavidus necator