Literature summary for 3.1.2.25 extracted from

Khandokar, Y.; Srivastava, P.; Sarker, S.; Swarbrick, C.; Aragao, D.; Cowieson, N.; Forwood, J.
Structural and functional characterization of the PaaI thioesterase from Streptococcus pneumoniae reveals a dual specificity for phenylacetyl-CoA and medium-chain fatty Acyl-CoAs and a novel CoA-induced fit mechanism (2016), J. Biol. Chem., 291, 1866-1876 .

Cloned(Commentary)

Cloned (Comment)	Organism
expressed in Escherichia coli BL21(DE3) cells	Streptococcus pneumoniae

Crystallization (Commentary)

Crystallization (Comment)	Organism
hanging drop vapor diffusion method. Crystals of wild type enzyme and mutants N37A, D52A, and T68A are obtained in conditions composed of 200 mM ammonium tartrate dibasic, 20% (w/v) PEG 3350, 100 mM Tris, pH 7.5; 200 mM lithium sulfate, 100 mM Tris-HCl, 30% (w/v) PEG 4000, 200 mM magnesium chloride, PEG 400, 100 mM HEPES, pH 7.5; 200 mM magnesium chloride, 14% (w/v) PEG 3350, 100 mM Tris, pH 6.0, respectively	Streptococcus pneumoniae

Crystallization (Comment)

Organism

hanging drop vapor diffusion method. Crystals of wild type enzyme and mutants N37A, D52A, and T68A are obtained in conditions composed of 200 mM ammonium tartrate dibasic, 20% (w/v) PEG 3350, 100 mM Tris, pH 7.5; 200 mM lithium sulfate, 100 mM Tris-HCl, 30% (w/v) PEG 4000, 200 mM magnesium chloride, PEG 400, 100 mM HEPES, pH 7.5; 200 mM magnesium chloride, 14% (w/v) PEG 3350, 100 mM Tris, pH 6.0, respectively

Streptococcus pneumoniae

Protein Variants

Protein Variants	Comment	Organism
D52A	the mutant has completely abolished activity	Streptococcus pneumoniae
N37A	the mutant has completely abolished activity	Streptococcus pneumoniae
T68A	the mutant shows a marked reduction in activity compared to the wild type enzyme	Streptococcus pneumoniae

Organism

Organism	UniProt	Comment	Textmining
Streptococcus pneumoniae	A0A0M3KL39	-	-
Streptococcus pneumoniae ATCC BAA-334	A0A0M3KL39	-	-

Purification (Commentary)

Purification (Comment)	Organism
HisTrap affinity column chromatography and gel filtration	Streptococcus pneumoniae

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
acetyl-CoA + H2O	very low activity	Streptococcus pneumoniae	acetate + CoA	-	?
acetyl-CoA + H2O	very low activity	Streptococcus pneumoniae ATCC BAA-334	acetate + CoA	-	?
arachidonyl-CoA + H2O	very low activity	Streptococcus pneumoniae	arachidonate + CoA	-	?
butyryl-CoA + H2O	low activity	Streptococcus pneumoniae	butanoate + CoA	-	?
butyryl-CoA + H2O	low activity	Streptococcus pneumoniae ATCC BAA-334	butanoate + CoA	-	?
decanoyl-CoA + H2O	highest activity	Streptococcus pneumoniae	decanoate + CoA	-	?
dodecanoyl-CoA + H2O	high activity	Streptococcus pneumoniae	dodecanoate + CoA	-	?
hexanoyl-CoA + H2O	-	Streptococcus pneumoniae	hexanoate + CoA	-	?
additional information	no activity with malonyl-CoA, palmitoyl-CoA, and stearoyl-CoA	Streptococcus pneumoniae	?	-	?
additional information	no activity with malonyl-CoA, palmitoyl-CoA, and stearoyl-CoA	Streptococcus pneumoniae ATCC BAA-334	?	-	?
myristoyl-CoA + H2O	very low activity	Streptococcus pneumoniae	myristate + CoA	-	?
myristoyl-CoA + H2O	very low activity	Streptococcus pneumoniae ATCC BAA-334	myristate + CoA	-	?
octanoyl-CoA + H2O	high activity	Streptococcus pneumoniae	octanoate + CoA	-	?
palmitoyl-CoA + H2O	-	Streptococcus pneumoniae	palmitate + CoA	-	?
palmitoyl-CoA + H2O	-	Streptococcus pneumoniae ATCC BAA-334	palmitate + CoA	-	?
phenylacetyl-CoA + H2O	low activity. The wild type enzyme displays a greater activity for phenylacetyl-CoA than very short and long chain saturated fatty acyl-CoA substrates	Streptococcus pneumoniae	phenylacetate + CoA	-	?

Subunits

Subunits	Comment	Organism
tetramer	-	Streptococcus pneumoniae

Synonyms

Synonyms	Comment	Organism
PaaI	-	Streptococcus pneumoniae
PaaI thioesterase	-	Streptococcus pneumoniae

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism	Structure
6.5	-	phenylacetyl-CoA	wild type enzyme, at pH 7.6 and 21°C	Streptococcus pneumoniae
32.8	-	decanoyl-CoA	wild type enzyme, at pH 7.6 and 21°C	Streptococcus pneumoniae

kcat/KM [mM/s]

kcat/KM Value [1/mMs^-1]	kcat/KM Value Maximum [1/mMs^-1]	Substrate	Comment	Organism	Structure
72	-	phenylacetyl-CoA	wild type enzyme, at pH 7.6 and 21°C	Streptococcus pneumoniae
180	-	decanoyl-CoA	wild type enzyme, at pH 7.6 and 21°C	Streptococcus pneumoniae