Literature summary for 5.3.3.1 extracted from

Hutchinson, M.H.; Morreale, G.; Middelberg, A.P.; Chase, H.A.
Production of enzymatically active ketosteroid isomerase following insoluble expression in Escherichia coli (2006), Biotechnol. Bioeng., 95, 724-733.

Cloned(Commentary)

Cloned (Comment)	Organism
expression of C-terminally His6-tagged enzyme as insoluble protein in inclusion bodies in Escherichia coli strain BLR(DE3)	Comamonas testosteroni

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
5-Androstene-3,17-dione	Comamonas testosteroni	-	4-Androstene-3,17-dione	-	?

Organism

Organism	UniProt	Comment	Textmining
Comamonas testosteroni	P00947	i.e. Pseudomonas testosteroni	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant C-terminally His6-tagged enzyme from Escherichia coli strain BLR(DE3) to homogeneity by expanded bed adsorption column utilizing immobilized metal affinity chromatography, this integrated process greatly simplifies the recovery and purification of inclusion body proteins by removing the need for mechanical cell disruption, repeated inclusion body centrifugation, and difficult clarification operations, chemical extraction is as effective as homogenization at releasing the inclusion body proteins from the bacterial cells, over 99% purity of the enzyme, method optimization and validation, overview	Comamonas testosteroni

Purification (Comment)

Organism

recombinant C-terminally His6-tagged enzyme from Escherichia coli strain BLR(DE3) to homogeneity by expanded bed adsorption column utilizing immobilized metal affinity chromatography, this integrated process greatly simplifies the recovery and purification of inclusion body proteins by removing the need for mechanical cell disruption, repeated inclusion body centrifugation, and difficult clarification operations, chemical extraction is as effective as homogenization at releasing the inclusion body proteins from the bacterial cells, over 99% purity of the enzyme, method optimization and validation, overview

Comamonas testosteroni

Renatured (Commentary)

Renatured (Comment)	Organism
dilution refolding of recombinant enzyme from inclusion bodies, denatured recombinant C-terminally His6-tagged enzyme 10fold with refolding buffer containing 50 mM potassium phosphate, pH 7.0, 4°C, incubation at 4°C for 12 h, urea 0.8 M	Comamonas testosteroni

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
5-Androstene-3,17-dione	-	Comamonas testosteroni	4-Androstene-3,17-dione	-	?

Synonyms

Synonyms	Comment	Organism
DELTA5-3-ketosteroid isomerase	-	Comamonas testosteroni
ketosteroid isomerase	-	Comamonas testosteroni
KSI	-	Comamonas testosteroni

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
30	-	assay at	Comamonas testosteroni

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
7	-	assay at	Comamonas testosteroni