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5.3.3.1: steroid DELTA-isomerase

This is an abbreviated version!
For detailed information about steroid DELTA-isomerase, go to the full flat file.

Word Map on EC 5.3.3.1

Reaction

a 3-oxo-DELTA5-steroid
=
a 3-oxo-DELTA4-steroid

Synonyms

3-Keto-DELTA5-steroid isomerase, 3-Ketosteroid DELTA5-->DELTA4-isomerase, 3-ketosteroid isomerase, 3-Oxo steroid DELTA4-DELTA5-isomerase, 3-Oxo-delta5 steroid isomerase, 3-oxo-DELTA5-steroid isomerase, 3-Oxosteroid DELTA4-DELTA5-isomerase, 3-Oxosteroid DELTA5-DELTA4-isomerase, 3-Oxosteroid isomerase, 3beta-HSD, 3beta-HSD/isomerase, 3beta-hydroxysteroid dehydrogenase/5-ene-4-ene isomerase, 3beta-hydroxysteroid dehydrogenase/DELTA5-DELTA4 isomerase, 3beta-hydroxysteroid dehydrogenase/isomerase, 3beta-hydroxysteroid dehydrogenase/isomerase type 1, 3beta-hydroxysteroid dehydrogenase/isomerase type 2, 3KSI, 5-Ene-4-ene isomerase, 5-Pregnene-3,20-dione isomerase, delta 5-3-ketosteroid isomerase, DELTA-3-ketosteroid isomerase, Delta-5-3-ketosteroid isomerase, DELTA5(or DELTA4)-3-keto steroid isomerase, DELTA5-3-keto steroid isomerase, DELTA5-3-ketosteroid isomerase, DELTA5-3-oxosteroid isomerase, DELTA5-ketosteroid isomerase, DELTA5-steroid isomerase, glutathione transferase A3-3, GST A3-3, Hydroxysteroid isomerase, Isomerase, steroid DELTA, ketosteroid isomerase, KSI, More, Steroid 5-->4-isomerase, Steroid isomerase, TI, TI-WT, type I 3beta-hydroxysteroid dehydrogenase/isomerase

ECTree

     5 Isomerases
         5.3 Intramolecular oxidoreductases
             5.3.3 Transposing C=C bonds
                5.3.3.1 steroid DELTA-isomerase

Purification

Purification on EC 5.3.3.1 - steroid DELTA-isomerase

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PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
15N labelled KSI is purified from Escherichia coli BL21 DE3 grown in minimal medium (M9) containing 15N ammonium chloride as the nitrogen source.
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3beta-hydroxy-5-ene steroid dehydrogenase/isomerase enzyme system
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3beta-hydroxy-5-ene-steroid dehydrogenase/steroid DELTA5-DELTA4-ene isomerase enzyme complex
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affinity chromatography
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ammonium sulfate precipitation and phenyl Sepharose 6 column chromatography
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copurifies with 3beta-hydroxysteroid oxidase activity
Nocardia rhodochrous
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DELTA5-3-beta-hydroxysteroid dehydrogenase-isomerase system
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enzyme complex of 3beta-hydroxy-5-ene-steroid dehydrogenase and steroid 5 to 4 isomerase
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glutathione-Sepharose column chromatography
modified enzyme in which the membrane-spanning domain, residues 283-310, of the enzyme protein is deleted in the cDNA is expressed by baculovirus in the cytosol instead of in the microsomes and mitochondria of the Sf9 cells
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mutant enzymes D36A/K37R, D241N, D257L, D258L and D265N are expressed in a baculovirus/Sf9 cell system
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mutant enzymes D40N and Y16F
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recombinant C-terminally His6-tagged enzyme from Escherichia coli strain BLR(DE3) to homogeneity by expanded bed adsorption column utilizing immobilized metal affinity chromatography with denaturant concentration in the feed stream from 8 to 0 M urea, chemical extraction from cells, method optimization and validation, overview
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
recombinant enzyme from Escherichia coli strain BLR(DE3) to homogeneity by nickel affinity chromatography after dilution refolding, method optimization, overview
recombinant type 1 isozyme, wild-type and mutant enzymes, from Spodoptera frugiperda Sf9 cells
recombinant wild-type and mutant enzymes from Escherichia coli strain DH5alpha to over 99% purity
the 3beta-hydroxysteroid dehydrogenase cannot be separated from the isomerase
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