Literature summary for 5.3.3.1 extracted from

Sigala, P.; Caaveiro, J.; Ringe, D.; Petsko, G.; Herschlag, D.
Hydrogen bond coupling in the ketosteroid isomerase active site (2009), Biochemistry, 48, 6932-6939.

Cloned(Commentary)

Cloned (Comment)	Organism
expression of wild-type and mutant enzymes in Escherichia coli strain DH5alpha	Comamonas testosteroni
expression of wild-type and mutant enzymes in Escherichia coli strain DH5alpha	Pseudomonas putida

Crystallization (Commentary)

Crystallization (Comment)	Organism
mutant D103N/D40N bound to inhibitor equilenin, hanging drop vapor diffusion method, 0.002 ml of 25 mg/ml protein with equilenin in a molar ratio of 1:1.2 in 40 mM potassium phosphate, pH 7.2, are mixed with 0.002 ml of reservoir solution conraining 1.4 M ammonium sulfate, and 6.5% v/v 2-propanol, pH 7.0, room temperature, 1 week, X-ray diffraction structure determination and analysis at 1.1 A resolution	Pseudomonas putida

Crystallization (Comment)

Organism

mutant D103N/D40N bound to inhibitor equilenin, hanging drop vapor diffusion method, 0.002 ml of 25 mg/ml protein with equilenin in a molar ratio of 1:1.2 in 40 mM potassium phosphate, pH 7.2, are mixed with 0.002 ml of reservoir solution conraining 1.4 M ammonium sulfate, and 6.5% v/v 2-propanol, pH 7.0, room temperature, 1 week, X-ray diffraction structure determination and analysis at 1.1 A resolution

Pseudomonas putida

Protein Variants

Protein Variants	Comment	Organism
D103N/D40N	site-directed mutagenesis, 3,4,5-trifluorophenol bind as ionized phenolate to KSI containing the D103N mutation	Pseudomonas putida
Y57F/D40N	site-directed mutagenesis, 3,4,5-trifluorophenol bind as ionized phenolate to KSI containing the Y57F mutation	Comamonas testosteroni

Inhibitors

Inhibitors	Comment	Organism	Structure
equilenin	reaction intermediate analogue, binds to the active site, binding structure, overview	Comamonas testosteroni
equilenin	reaction intermediate analogue, binds to the active site, binding structure, overview	Pseudomonas putida

Organism

Organism	UniProt	Comment	Textmining
Comamonas testosteroni	-	-	-
Pseudomonas putida	P07445	-	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant wild-type and mutant enzymes from Escherichia coli strain DH5alpha to over 99% purity	Comamonas testosteroni
recombinant wild-type and mutant enzymes from Escherichia coli strain DH5alpha to over 99% purity	Pseudomonas putida

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
additional information	mechanism of KSI-catalyzed steroid isomerization, direct detection of physical coupling between the Y16 and D103 hydrogen bonds, ligand binding, overview	Pseudomonas putida	?	-	?
additional information	mechanism of KSI-catalyzed steroid isomerization, overview	Comamonas testosteroni	?	-	?

Synonyms

Synonyms	Comment	Organism
ketosteroid isomerase	-	Comamonas testosteroni
ketosteroid isomerase	-	Pseudomonas putida
KSI	-	Comamonas testosteroni
KSI	-	Pseudomonas putida