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Literature summary for 2.3.1.253 extracted from

  • Zha, W.; Rubin-Pitel, S.B.; Zhao, H.
    Characterization of the substrate specificity of PhlD, a type III polyketide synthase from Pseudomonas fluorescens (2006), J. Biol. Chem., 281, 32036-32047.
    View publication on PubMed
Search for more literature for 2.3.1.253

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli BL21(DE3) cells Pseudomonas fluorescens

Protein Variants

Protein Variants Comment Organism
A185D inactive Pseudomonas fluorescens
A60L the mutant still retains the wild type activity to react with lauroyl-CoA Pseudomonas fluorescens
H24L the mutant shows reduced reactivity toward lauroyl-CoA and has lost the ability to produce phloroglucinol Pseudomonas fluorescens
H24V the mutation significantly reduces the reactivity of the enzyme with lauroyl-CoA while still retaining its physiological activity to synthesize phloroglucinol Pseudomonas fluorescens
L59M the mutation significantly reduces the reactivity of the enzyme with lauroyl-CoA while still retaining its physiological activity to synthesize phloroglucinol Pseudomonas fluorescens
M21I the mutation significantly reduces the reactivity of the enzyme with lauroyl-CoA while still retaining its physiological activity to synthesize phloroglucinol Pseudomonas fluorescens
M21T the mutant still retains the wild type activity to react with lauroyl-CoA Pseudomonas fluorescens

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.013
-
 malonyl-CoA at pH 7.5 and 37°C Pseudomonas fluorescens

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
3 malonyl-CoA Pseudomonas fluorescens preferred substrate phloroglucinol + 3 CO2 + 3 CoA
-
 ?
3 malonyl-CoA Pseudomonas fluorescens Pf-5 preferred substrate phloroglucinol + 3 CO2 + 3 CoA
-
 ?

Organism

Organism UniProt Comment Textmining
Pseudomonas fluorescens Q4K418
-
-
Pseudomonas fluorescens Pf-5 Q4K418
-
-

Purification (Commentary)

Purification (Comment) Organism
Ni-NTA agarose resin column chromatography Pseudomonas fluorescens

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3 malonyl-CoA preferred substrate Pseudomonas fluorescens phloroglucinol + 3 CO2 + 3 CoA
-
 ?
3 malonyl-CoA preferred substrate Pseudomonas fluorescens Pf-5 phloroglucinol + 3 CO2 + 3 CoA
-
 ?
acetoacetyl-CoA 1.6% activity compared to malonyl-CoA Pseudomonas fluorescens phloroglucinol + 4-hydroxy-6-methyl-2H-pyran-2-one + CO2 + CoA
-
 ?
acetoacetyl-CoA 1.6% activity compared to malonyl-CoA Pseudomonas fluorescens Pf-5 phloroglucinol + 4-hydroxy-6-methyl-2H-pyran-2-one + CO2 + CoA
-
 ?
butyryl-CoA 0.2% activity compared to malonyl-CoA Pseudomonas fluorescens phloroglucinol + 4-hydroxy-6-propyl-2H-pyran-2-one + CO2 + CoA
-
 ?
butyryl-CoA 0.2% activity compared to malonyl-CoA Pseudomonas fluorescens Pf-5 phloroglucinol + 4-hydroxy-6-propyl-2H-pyran-2-one + CO2 + CoA
-
 ?
decanoyl-CoA 18% activity compared to malonyl-CoA Pseudomonas fluorescens 4-hydroxy-6-octyl-2H-pyran-2-one + 4-hydroxy-6-(2-oxoundecyl)-2H-pyran-2-one + 4-hydroxyl-6-(2',4',6'-trioxopentadecyl)-2-pyrone + CO2 + CoA
-
 ?
decanoyl-CoA 18% activity compared to malonyl-CoA Pseudomonas fluorescens Pf-5 4-hydroxy-6-octyl-2H-pyran-2-one + 4-hydroxy-6-(2-oxoundecyl)-2H-pyran-2-one + 4-hydroxyl-6-(2',4',6'-trioxopentadecyl)-2-pyrone + CO2 + CoA
-
 ?
hexanoyl-CoA 8% activity compared to malonyl-CoA Pseudomonas fluorescens 4-hydroxy-6-pentyl-2H-pyran-2-one + 4-hydroxy-6-(2-oxoheptyl)-2H-pyran-2-one + CO2 + CoA
-
 ?
lauroyl-CoA 7.6% activity compared to malonyl-CoA Pseudomonas fluorescens 6-decyl-4-hydroxy-2H-pyran-2-one + 4-hydroxy-6-(2-oxotridecyl)-2H-pyran-2-one + CO2 + CoA
-
 ?
additional information the enzyme shows barely detectable activity toward acetyl-CoA, isobutyryl-CoA, benzoyl-CoA, and crotonyl-CoA. No activity with myristoyl-CoA and palmitoyl-CoA Pseudomonas fluorescens ?
-
 ?
additional information the enzyme shows barely detectable activity toward acetyl-CoA, isobutyryl-CoA, benzoyl-CoA, and crotonyl-CoA. No activity with myristoyl-CoA and palmitoyl-CoA Pseudomonas fluorescens Pf-5 ?
-
 ?
octanoyl-CoA 7% activity compared to malonyl-CoA Pseudomonas fluorescens 6-heptyl-4-hydroxy-2H-pyran-2-one + 4-hydroxy-6-(2-oxononyl)-2H-pyran-2-one + 4-hydroxyl-6-(2',4',6'-trioxotridecyl)-2-pyrone + 4-hydroxyl-6-(2',4',6',8'-tetraoxopentadecyl)-2-pyrone + CO2 + CoA
-
 ?
phenylacetyl-CoA 3.3% activity compared to malonyl-CoA Pseudomonas fluorescens 6-(cyclohepta-1,3,6-trien-1-yl)-4-hydroxy-2H-pyran-2-one + 6-[2-(cyclohepta-1,3,6-trien-1-yl)-2-oxoethyl]-4-hydroxy-2H-pyran-2-one + CO2 + CoA
-
 ?

Synonyms

Synonyms Comment Organism
PhlD
-
 Pseudomonas fluorescens

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
25 37 the times required to reduce the activity of the enzyme by 50% at 25, 30, and 37°C are 128, 74, and 7.2 min, respectively Pseudomonas fluorescens

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.4
-
 malonyl-CoA at pH 7.5 and 37°C Pseudomonas fluorescens

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
31.38
-
 malonyl-CoA at pH 7.5 and 37°C Pseudomonas fluorescens