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Literature summary for 3.7.1.4 extracted from

  • Han, J.T.; Zhang, S.P.; Jia, W.J.; Zhang, Z.; Wang, Y.; He, Y.X.
    Discovery and structural analysis of a phloretin hydrolase from the opportunistic human pathogen Mycobacterium abscessus (2019), FEBS J., 286, 1959-1971 .
    View publication on PubMed
Search for more literature for 3.7.1.4

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli BL21(DE3) cells Mycobacteroides abscessus

Crystallization (Commentary)

Crystallization (Comment) Organism
hanging drop vapor diffusion method, using 100 mM Na cacodylate pH 5.0, 1.0 M lithium chloride, 6.8% (w/v) polyethylene glycol 6000 Mycobacteroides abscessus

Protein Variants

Protein Variants Comment Organism
A213S the mutant completely loses catalytic activity towards phloretin Mycobacteroides abscessus
E164A the mutation completely abolishes the hydrolytic activity towards phloretin Mycobacteroides abscessus
E273A the mutation completely abolishes the hydrolytic activity towards phloretin Mycobacteroides abscessus
H118A the mutant completely loses catalytic activity towards phloretin Mycobacteroides abscessus
H133A the kcat value for phloretin is decreased at least by 1 order of magnitude, whereas the corresponding Km value is barely affected compared to the wild type enzyme Mycobacteroides abscessus
H160A the mutant completely loses catalytic activity towards phloretin Mycobacteroides abscessus
H160F the mutant completely loses catalytic activity towards phloretin Mycobacteroides abscessus
H217A the mutant has a Km value increased by 3fold and a kcat value decreased by 6fold compared to the wild type enzyme Mycobacteroides abscessus
H269A the kcat value for phloretin is decreased at least by 1 order of magnitude, whereas the corresponding Km value is barely affected compared to the wild type enzyme Mycobacteroides abscessus
I162A the mutant completely loses catalytic activity towards phloretin Mycobacteroides abscessus
Q266F the mutant completely loses catalytic activity towards phloretin Mycobacteroides abscessus
Y125A the mutant completely loses catalytic activity towards phloretin Mycobacteroides abscessus
Y232A the Km value for phloretin increases by 3fold and the kcat value decreases by 5fold compared to the wild type enzyme Mycobacteroides abscessus

Inhibitors

Inhibitors Comment Organism Structure
2,4-diacetylphloroglucinol competitive inhibition Mycobacteroides abscessus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0572
-
 phloretin mutant enzyme H133A, at pH 6.5 and 25°C Mycobacteroides abscessus
0.0812
-
 phloretin wild type enzyme, at pH 6.5 and 25°C Mycobacteroides abscessus
0.0894
-
 phloretin mutant enzyme H269A, at pH 6.5 and 25°C Mycobacteroides abscessus
0.26
-
 phloretin mutant enzyme Y232A, at pH 6.5 and 25°C Mycobacteroides abscessus
0.289
-
 phloretin mutant enzyme H217A, at pH 6.5 and 25°C Mycobacteroides abscessus
2.59
-
 monoacetylphloroglucinol wild type enzyme, at pH 6.5 and 25°C Mycobacteroides abscessus

Metals/Ions

Metals/Ions Comment Organism Structure
Zn2+ contains zinc Mycobacteroides abscessus

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
monoacetylphloroglucinol + H2O Mycobacteroides abscessus
-
 ?
-
 ?
monoacetylphloroglucinol + H2O Mycobacteroides abscessus 103
-
 ?
-
 ?
phloretin + H2O Mycobacteroides abscessus
-
 phloroglucinol + 3-(4-hydroxyphenyl)propionic acid
-
 ?
phloretin + H2O Mycobacteroides abscessus 103
-
 phloroglucinol + 3-(4-hydroxyphenyl)propionic acid
-
 ?

Organism

Organism UniProt Comment Textmining
Mycobacteroides abscessus
-
-
-
Mycobacteroides abscessus 103
-
-
-

Purification (Commentary)

Purification (Comment) Organism
Ni-NTA column chromatography and Superdex S200 gel filtration Mycobacteroides abscessus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
monoacetylphloroglucinol + H2O
-
 Mycobacteroides abscessus ?
-
 ?
monoacetylphloroglucinol + H2O
-
 Mycobacteroides abscessus 103 ?
-
 ?
phloretin + H2O
-
 Mycobacteroides abscessus phloroglucinol + 3-(4-hydroxyphenyl)propionic acid
-
 ?
phloretin + H2O
-
 Mycobacteroides abscessus 103 phloroglucinol + 3-(4-hydroxyphenyl)propionic acid
-
 ?

Synonyms

Synonyms Comment Organism
Phlg
-
 Mycobacteroides abscessus

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.05
-
 phloretin mutant enzyme H269A, at pH 6.5 and 25°C Mycobacteroides abscessus
0.07
-
 phloretin mutant enzyme H133A, at pH 6.5 and 25°C Mycobacteroides abscessus
0.13
-
 phloretin mutant enzyme H217A, at pH 6.5 and 25°C Mycobacteroides abscessus
0.16
-
 phloretin mutant enzyme Y232A, at pH 6.5 and 25°C Mycobacteroides abscessus
0.85
-
 phloretin wild type enzyme, at pH 6.5 and 25°C Mycobacteroides abscessus
49.6
-
 monoacetylphloroglucinol wild type enzyme, at pH 6.5 and 25°C Mycobacteroides abscessus

Ki Value [mM]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
0.0116
-
 2,4-diacetylphloroglucinol wild type enzyme, with phloretin as substrate, at pH 6.5 and 25°C Mycobacteroides abscessus
0.0453
-
 2,4-diacetylphloroglucinol wild type enzyme, with monoacetylphloroglucinol as substrate, at pH 6.5 and 25°C Mycobacteroides abscessus

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.45
-
 phloretin mutant enzyme H217A, at pH 6.5 and 25°C Mycobacteroides abscessus
0.56
-
 phloretin mutant enzyme H269A, at pH 6.5 and 25°C Mycobacteroides abscessus
0.62
-
 phloretin mutant enzyme Y232A, at pH 6.5 and 25°C Mycobacteroides abscessus
1.2
-
 phloretin mutant enzyme H133A, at pH 6.5 and 25°C Mycobacteroides abscessus
10.5
-
 phloretin wild type enzyme, at pH 6.5 and 25°C Mycobacteroides abscessus