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Literature summary for 3.7.1.24 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 3.7.1.24

Cloned(Commentary)

Cloned (Comment) Organism
gene phlg, sequence comparison Pseudomonas fluorescens

Protein Variants

Protein Variants Comment Organism
E160A site-directed mutagenesis, almost inactive enzyme Pseudomonas fluorescens
E274A site-directed mutagenesis, a mutant of very low activity Pseudomonas fluorescens
H214A site-directed mutagenesis, the mutant shows an unaltered KM but reduced catalytic efficiency with diacetyl phloroglucinol compared to the wild-type Pseudomonas fluorescens
H270A site-directed mutagenesis, inactive enzyme Pseudomonas fluorescens
additional information mutation of His123, His251, Glu154 and Glu255 (conserved zinc binding residues) results in variants that are 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 15fold reduced activity and an inactive variant Pseudomonas fluorescens
Y121A site-directed mutagenesis, the mutant shows reduced KM and catalytic efficiency with diacetyl phloroglucinol compared to the wild-type Pseudomonas fluorescens

Metals/Ions

Metals/Ions Comment Organism Structure
Zn2+ a Zn-dependent C-C hydrolase, bound in the active site, chelated by four amino acid side-chains, His270, His129, Glu274 and Glu160, might also have structural role in Phlg Pseudomonas fluorescens

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2,4-diacetylphloroglucinol + H2O Pseudomonas fluorescens
-
 2-acetylphloroglucinol + acetate
-
 ?

Organism

Organism UniProt Comment Textmining
Pseudomonas fluorescens Q4K423
-
-

Reaction

Reaction Comment Organism Reaction ID
2,4-diacetylphloroglucinol + H2O = 2-acetylphloroglucinol + acetate catalytic reaction mechanism, overview. A water molecule, bound by the zinc, is responsible for initiating the CĀ–C bond cleavage reaction through nucleophilic attack at the acyl carbonyl carbon atom Pseudomonas fluorescens
a

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2,4-diacetylphloroglucinol + H2O
-
 Pseudomonas fluorescens 2-acetylphloroglucinol + acetate
-
 ?
2,4-diacetylphloroglucinol + H2O modeling of a theoretical transition state for the substrate hydrolysis into the active site revealing that a water molecule, bound by the zinc, can be responsible for initiating the C-C bond cleavage reaction through nucleophilic attack at the acyl carbonyl carbon atom. Residue Asn132 in Phlg is thought to make a hydrogen bond to the acyl group of the native substrate in the enzyme. Tyr121 is thought to hydrogen bond to the phenol group in the 1-position of the aromatic ring between the two acetyl groups in the substrate Pseudomonas fluorescens 2-acetylphloroglucinol + acetate
-
 ?

Synonyms

Synonyms Comment Organism
diacetyl phloroglucinol hydrolase
-
 Pseudomonas fluorescens
diacetylphloroglucinol hydrolase
-
 Pseudomonas fluorescens
Phlg
-
 Pseudomonas fluorescens

General Information

General Information Comment Organism
evolution the C-C hydrolase enzyme exhibits a Bet v1-like fold rather than the alpha/beta hydrolase fold common to C-C hydrolases Pseudomonas fluorescens
malfunction mutation of His123, His251, Glu154 and Glu255 (conserved zinc binding residues) results in variants that are 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 15fold reduced activity and an inactive variant Pseudomonas fluorescens
additional information the enzyme structure, PDB ID 3HWP, is used for a homology model of phloretin hydrolase, Phy, EC 3.7.1.4, from Eubacterium ramulus. Active site structure of the enzyme Phlg, overview Pseudomonas fluorescens