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Literature summary 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.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
5.3.3.1 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)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
5.3.3.1 5-Androstene-3,17-dione Comamonas testosteroni
-
4-Androstene-3,17-dione
-
?

Organism

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

Purification (Commentary)

EC Number Purification (Comment) Organism
5.3.3.1 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)

EC Number Renatured (Comment) Organism
5.3.3.1 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)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
5.3.3.1 5-Androstene-3,17-dione
-
Comamonas testosteroni 4-Androstene-3,17-dione
-
?

Synonyms

EC Number Synonyms Comment Organism
5.3.3.1 DELTA5-3-ketosteroid isomerase
-
Comamonas testosteroni
5.3.3.1 ketosteroid isomerase
-
Comamonas testosteroni
5.3.3.1 KSI
-
Comamonas testosteroni

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
5.3.3.1 30
-
assay at Comamonas testosteroni

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
5.3.3.1 7
-
assay at Comamonas testosteroni