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Literature summary for 3.7.1.4 extracted from

  • Frank, A.; Siirola, E.; Kroutil, W.; Grogan, G.
    Mutational analysis of the C-C bond cleaving enzyme phloretin hydrolase from Eubacterium ramulus (2014), Topics Catal., 57, 376-384.
No PubMed abstract available
Search for more literature for 3.7.1.4

Cloned(Commentary)

Cloned (Comment) Organism
gene phy, DNA and amino acid sequence determination and analysis, recombinant expression of wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) Eubacterium ramulus

Protein Variants

Protein Variants Comment Organism
A126S site-directed mutagenesis Eubacterium ramulus
F218Y site-directed mutagenesis Eubacterium ramulus
additional information mutation of His123, His251, Glu154 and Glu255 (conserved zinc binding residues) results in variants that were either poorly expressed, or of much reduced activity. Mutation of Tyr115 and His203, thought to bind the phenol groups in the 1-and 3-positions of the phloroglucinol ring respectively, results in variants of 15-fold reduced activity and an inactive variant Eubacterium ramulus

Metals/Ions

Metals/Ions Comment Organism Structure
Zn2+ a Zn-dependent C-C hydrolase, His123, His251, Glu154 and Glu255 are conserved zinc binding residues Eubacterium ramulus

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
30000
-
 2 * 30000, recombinant enzyme, SDS-PAGE Eubacterium ramulus
55000
-
 recombinant enzyme, gel filtration Eubacterium ramulus

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
phloretin + H2O Eubacterium ramulus
-
 phloretate + phloroglucinol
-
 ?

Organism

Organism UniProt Comment Textmining
Eubacterium ramulus Q715L4 Eubacterium ramulus is part of the natural gut flora in humans, gene phy
-

Purification (Commentary)

Purification (Comment) Organism
recombinant wild-type and mutant enzymes from Escherichia coli strain Bl21(DE3) Eubacterium ramulus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
additional information no activity with 1,3-diphenyl 1,3-propanedione and diacetylphloroglucinol, a substrate of diacetyl phloroglucinol hydrolase, Phlg Eubacterium ramulus ?
-
 ?
phloretin + H2O
-
 Eubacterium ramulus phloretate + phloroglucinol
-
 ?
phloretin + H2O phloretin hydrolase catalyzes the hydrolysis of the dihydrochalcone phloretin to phloroglucinol and phloretic acid, performing a formal retro-Friedel-Crafts acylation reaction on its substrate, it is a C-C bond cleaving enzyme Eubacterium ramulus phloretate + phloroglucinol
-
 ?

Subunits

Subunits Comment Organism
homodimer 2 * 30000, recombinant enzyme, SDS-PAGE Eubacterium ramulus

Synonyms

Synonyms Comment Organism
Phy
-
 Eubacterium ramulus

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
30
-
 assay at Eubacterium ramulus

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
 assay at Eubacterium ramulus

General Information

General Information Comment Organism
additional information comparison of the enzyme phloretin hydrolase Phy from Eubacterium ramulus with the diacetyl phloroglucinol hydrolase (Phlg) from Pseudomonas fluorescens, which catalyses a similar, hydrolytic, de-acylation of its substrate, homology modeling of Phy based on the structure of Phlg, PDB ID 3HWP Eubacterium ramulus
physiological function the enzyme catalyses the degradation of plant-derived dihydrochalcone phloretin Eubacterium ramulus