Literature summary 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.

Cloned(Commentary)

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

Protein Variants

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

KM Value [mM]

EC Number	KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism	Structure
2.3.1.253	0.013	-	malonyl-CoA	at pH 7.5 and 37°C	Pseudomonas fluorescens

Natural Substrates/ Products (Substrates)

EC Number	Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
2.3.1.253	3 malonyl-CoA	Pseudomonas fluorescens	preferred substrate	phloroglucinol + 3 CO2 + 3 CoA	-	?
2.3.1.253	3 malonyl-CoA	Pseudomonas fluorescens Pf-5	preferred substrate	phloroglucinol + 3 CO2 + 3 CoA	-	?

Organism

EC Number	Organism	UniProt	Comment	Textmining
2.3.1.253	Pseudomonas fluorescens	Q4K418	-	-
2.3.1.253	Pseudomonas fluorescens Pf-5	Q4K418	-	-

Purification (Commentary)

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

Substrates and Products (Substrate)

EC Number	Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
2.3.1.253	3 malonyl-CoA	preferred substrate	Pseudomonas fluorescens	phloroglucinol + 3 CO2 + 3 CoA	-	?
2.3.1.253	3 malonyl-CoA	preferred substrate	Pseudomonas fluorescens Pf-5	phloroglucinol + 3 CO2 + 3 CoA	-	?
2.3.1.253	acetoacetyl-CoA	1.6% activity compared to malonyl-CoA	Pseudomonas fluorescens	phloroglucinol + 4-hydroxy-6-methyl-2H-pyran-2-one + CO2 + CoA	-	?
2.3.1.253	acetoacetyl-CoA	1.6% activity compared to malonyl-CoA	Pseudomonas fluorescens Pf-5	phloroglucinol + 4-hydroxy-6-methyl-2H-pyran-2-one + CO2 + CoA	-	?
2.3.1.253	butyryl-CoA	0.2% activity compared to malonyl-CoA	Pseudomonas fluorescens	phloroglucinol + 4-hydroxy-6-propyl-2H-pyran-2-one + CO2 + CoA	-	?
2.3.1.253	butyryl-CoA	0.2% activity compared to malonyl-CoA	Pseudomonas fluorescens Pf-5	phloroglucinol + 4-hydroxy-6-propyl-2H-pyran-2-one + CO2 + CoA	-	?
2.3.1.253	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	-	?
2.3.1.253	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	-	?
2.3.1.253	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	-	?
2.3.1.253	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	-	?
2.3.1.253	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	?	-	?
2.3.1.253	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	?	-	?
2.3.1.253	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	-	?
2.3.1.253	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

EC Number	Synonyms	Comment	Organism
2.3.1.253	PhlD	-	Pseudomonas fluorescens

Temperature Stability [°C]

EC Number	Temperature Stability Minimum [°C]	Temperature Stability Maximum [°C]	Comment	Organism
2.3.1.253	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]

EC Number	Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism	Structure
2.3.1.253	0.4	-	malonyl-CoA	at pH 7.5 and 37°C	Pseudomonas fluorescens

kcat/KM [mM/s]

EC Number	kcat/KM Value [1/mMs^-1]	kcat/KM Value Maximum [1/mMs^-1]	Substrate	Comment	Organism	Structure
2.3.1.253	31.38	-	malonyl-CoA	at pH 7.5 and 37°C	Pseudomonas fluorescens

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Release 2023.1 (January 2023)