Literature summary for 2.3.1.151 extracted from

Klundt, T.; Bocola, M.; Luetge, M.; Beuerle, T.; Liu, B.; Beerhues, L.
A single amino acid substitution converts benzophenone synthase into phenylpyrone synthase (2009), J. Biol. Chem., 284, 30957-30964.

Search for more literature for 2.3.1.151

Cloned(Commentary)

Cloned (Comment)	Organism
wild-type benzophenone synthase and enzyme mutants are heterologously expressed as N-terminally His6-tagged proteins in Escherichia coli	Hypericum androsaemum

Protein Variants

Protein Variants	Comment	Organism
T135A	inactive enzyme	Hypericum androsaemum
T135F	mutant functionally resemble the wild-type enzyme, albeit with reduced catalytic activities	Hypericum androsaemum
T135G	inactive enzyme	Hypericum androsaemum
T135I	inactive enzyme	Hypericum androsaemum
T135L	a point mutation in the active site cavity transforms benzophenone synthase into a functional phenylpyrone synthase (PPS). The dramatic change in both substrate and product specificities of benzophenone synthase is rationalized by homology modeling. The mutation may open a new pocket that accommodates the phenyl moiety of the triketide intermediate but limits polyketide elongation to two reactions, resulting in phenylpyrone formation. kcat (substrate: benzoyl-CoA): 0.0086/sec, Km (benzoyl-CoA): 0.001 mM, Km (malonyl-CoA): 8.7 mM. In contrast to the wild-type enzyme, the T135L mutant forms phenylpyrone as a major product and only traces of 2,4,6-trihydroxybenzophenone when incubated with benzoyl-CoA and malonyl-CoA. T135L mutant is almost inactive with 3-hydroxybenzoyl-CoA	Hypericum androsaemum
T135N	inactive enzyme	Hypericum androsaemum
T135S	mutant functionally resemble the wild-type enzyme, albeit with reduced catalytic activities	Hypericum androsaemum
T135V	inactive enzyme	Hypericum androsaemum
T135Y	inactive enzyme	Hypericum androsaemum

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism	Structure
0.001	-	benzoyl-CoA	mutant T135L	Hypericum androsaemum
0.0086	-	benzoyl-CoA	wild-type	Hypericum androsaemum
0.0087	-	malonyl-CoA	mutant T135L	Hypericum androsaemum
0.031	-	malonyl-CoA	wild-type	Hypericum androsaemum

Molecular Weight [Da]

Molecular Weight [Da]	Molecular Weight Maximum [Da]	Comment	Organism
43000	-	SDS-PAGE, molecular weight of mutant T135L	Hypericum androsaemum

Organism

Organism	UniProt	Comment	Textmining
Hypericum androsaemum	-	-	-

Purification (Commentary)

Purification (Comment)	Organism
purified on a nickel-nitrilotriacetic acid affinity matrix	Hypericum androsaemum

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
2 malonyl-CoA + benzoyl-CoA	relative activity: 100%, wildtype benzophenone synthase forms 2,4,6-trihydroxybenzophenone as a major product when incubated with benzoyl-CoA and malonyl-CoA, along with small amounts of 6-phenyl-4-hydroxy-2-pyrone	Hypericum androsaemum	3 CoA + 2,4,6-trihydroxybenzophenone + 2 CO2	-	?
3 malonyl-CoA + 3-hydroxybenzoyl-CoA	relative activity: 57%	Hypericum androsaemum	4 CoA + 2,3',4,6-tetrahydroxybenzophenone + 3 CO2	-	?
3 malonyl-CoA + benzoyl-CoA	relative activity: 100%, wildtype benzophenone synthase forms 2,4,6-trihydroxybenzophenone as a major product when incubated with benzoyl-CoA and malonyl-CoA, along with small amounts of 6-phenyl-4-hydroxy-2-pyrone	Hypericum androsaemum	4 CoA + 6-phenyl-4-hydroxy-2-pyrone + 3 CO2	-	?

Synonyms

Synonyms	Comment	Organism
benzophenone synthase	-	Hypericum androsaemum
BPS	-	Hypericum androsaemum

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
30	-	assay at	Hypericum androsaemum

Temperature Range [°C]

Temperature Minimum [°C]	Temperature Maximum [°C]	Comment	Organism
35	40	optimal temperature	Hypericum androsaemum

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism	Structure
0.00685	-	benzoyl-CoA	mutant T135L	Hypericum androsaemum
0.055	-	benzoyl-CoA	wild-type	Hypericum androsaemum

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
7	-	assay at	Hypericum androsaemum
7.5	-	optimal pH	Hypericum androsaemum

kcat/KM [mM/s]

kcat/KM Value [1/mMs^-1]	kcat/KM Value Maximum [1/mMs^-1]	Substrate	Comment	Organism	Structure
6.414	-	benzoyl-CoA	wild-type	Hypericum androsaemum
6.833	-	benzoyl-CoA	mutant T135L	Hypericum androsaemum

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Release 2023.1 (January 2023)