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1.3.99.4: 3-oxosteroid 1-dehydrogenase

This is an abbreviated version!
For detailed information about 3-oxosteroid 1-dehydrogenase, go to the full flat file.

Word Map on EC 1.3.99.4

Reaction

a 3-oxosteroid
+
acceptor
=
a 3-oxo-DELTA1-steroid
 +
reduced acceptor

Synonyms

1-ene-dehydrogenase, 3-keto-5alpha-steroid DELTA(1)-dehydrogenase, 3-ketosteroid 1(2)-dehydrogenase, 3-ketosteroid delta1-dehydrogenase, 3-ketosteroid-1-en-dehydrogenase, 3-ketosteroid-D1-dehydrogenase, 3-ketosteroid-DELTA1-dehydrogenase, 3-ketosteroid-DELTA1-dehydrogenases, 3-oxosteroid D1-dehydrogenase, 3-oxosteroid Delta1-dehydrogenase, 3-oxosteroid:(2,6-dichlorphenolindophenol) DELTA1-oxidoreductase, 3-oxosteroid:(2,6-dichlorphenolindophenol)D1-oxidoreductase, 4-en-3-oxosteroid:(acceptor)-1-en-oxido-reductase, 4-ene-3-oxosteroid: (acceptor)-1-ene-oxoreductase, 4-ene-3-oxosteroid:(acceptor)l-ene-oxidoreductase, AcmB, AcmB2, cholest-4-en-3-one DELTA1-dehydrogenase, D1-dehydrogenase, D1-steroid reductase, dehydrogenase, 3-oxo steroid Delta 1-, Delta1-dehydrogenase, DELTA1-KstD, DELTA1-KSTD1, DELTA1-KstDA, DELTA1-KstDR, DELTA1-steroid reductase, ketosteroid DELTA1-dehydrogenase, KsdD, KsdD1, KsdD2, KsdD3, KsdD5, KsdDM, KSDH, KstD, KSTD1, KSTD2, KSTD3, KSTD3 orthologue, KstDF, MnKstD1, MnKstD2, MsKstD1, MsKstD2, MsKstD3, MsKstD4, ReKstD1, Rv3537, steroid-1-dehydrogenase

ECTree

     1 Oxidoreductases
         1.3 Acting on the CH-CH group of donors
             1.3.99 With unknown physiological acceptors
                1.3.99.4 3-oxosteroid 1-dehydrogenase

Cloned

Cloned on EC 1.3.99.4 - 3-oxosteroid 1-dehydrogenase

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CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
expressed in Bacillus subtilis strain WB600
-
expressed in Escherichia coli BL21(DE3) cells
expression in Escherichia coli
expression in Streptomyces lividans
-
gene acmB, a synthetic DELTA1-KstD, developed based on the AcmB gene of Sterolibacterium denitrificans, stable recombinant expression in Escherichia coli and in human Hep-3B and U-937 cells. The enzyme, when expressed in human cells, introduces a double bond between the C-1 and C-2 atoms of 3-oxosteroids. This represents an integral step needed for cholestane A-ring aromatization and B-ring opening for which humans have no known orthologue, coexpression with cholesterol-3-hydroxy-dehydrogenase (CholD) and 3-oxosteroid-9alpha-hydroxylase (Kst-9alphaH)
gene acmB, recombinant expression in Escherichia coli
gene amcB, recombinant enzyme expression in Escherichia coli
gene ksdD, DNA and amino acid sequence determination and analysis, phylogenetic analysis and tree, recombinant overexpression of His-tagged enzyme in Escherichia coli strain BL21(DE3)
gene ksdD, phylogenetic analysis, expression in Bacillus subtilis strain 168 with 18fold higher activity compared to wild-type Mycobacterium neoaurum strain JC-12
-
gene kstd, DNA and amino acid sequence determination and analysis, phylogenetic analysis and tree, recombinant overexpression of His-tagged enzyme in Escherichia coli strain BL21(DE3)
gene kstD, DNA and amino acid sequence determination and analysis, phylogenetic tree, heterologous expression of the His-tagged enzyme, by low-temperature induction at 22°C, the recombinant protein appears partly as soluble protein
gene kstD, DNA and amino acid sequence determination and analysis, recombinant expression in Escherichia coli strain XL10-Gold
gene kstD1, a synthetic DELTA1-KstD, developed based on the KstD1 gene of Rhodococcus erythropolis, stable recombinant expression of the enzyme as an N-terminal FLAG His-patch thioredoxin fusion protein in Escherichia coli and in human Hep-3B and U-937 cells. The enzyme, when expressed in human cells, introduces a double bond between the C-1 and C-2 atoms of 3-oxosteroids. This represents an integral step needed for cholestane A-ring aromatization and B-ring opening for which humans have no known orthologue, coexpression with cholesterol-3-hydroxy-dehydrogenase (CholD) and 3-oxosteroid-9alpha-hydroxylase (Kst-9alphaH)
gene kstD1, heterologous expression of the His-tagged DELTA1-KSTD1 protein in Escherichia coli strain BL21(DE3)
gene kstD1, recombinant enzyme expression in Escherichia coli strain BL21(DE3)
gene kstd2, DNA and amino acid sequence determination and analysis, phylogenetic analysis and tree, recombinant overexpression of His-tagged enzyme in Escherichia coli strain BL21(DE3)
gene KSTD2, DNA and amino acid sequence determination and analysis, recombinant expression of N-terminally or C-terminally His-tagged enzyme in Escherichia coli strain BL21(DE3)
gene kstD3, cloning and expression as a soluble protein in Escherichia coli
-
gene kstd3, DNA and amino acid sequence determination and analysis, phylogenetic analysis and tree, recombinant overexpression of His-tagged enzyme in Escherichia coli strain BL21(DE3)
gene kstDF, DNA and amino acid sequence determination, analysis, and comparison, expression in Escherichia coli strain BL21(lambdaDE3) and in Pichia pastoris strains KM71 and GS115 cytoplasm, both intracellularly and extracelluarly, the latter is inactive, method optimization
-
into the pJam2 expression vector under the acetamidase promoter and introduced into the deltaksdD-1 mutant
-
isozyme KsdD3, sequence comparisons and phylogenetic analysis, genetic organization of the five KsdD homologues and in silico analysis of ksdD promoter regions, the putative promoter sequence for ksdD3 ORF lies in the intergenic region between bphE3 and ksdD3 ORFs, quantitative real time PCR enzyme expression analysis, recombinant expression of codon-optimized His-tagged enzyme in Escherichia coli strain BL21(DE3)
-
isozyme KsdD5, sequence comparisons and phylogenetic analysis, genetic organization of the five KsdD homologues in silico analysis of ksdD promoter regions, quantitative real time PCR enzyme expression analysis, recombinant expression of codon-optimized His-tagged enzyme in Escherichia coli strain BL21(DE3)
-
isozyme KstD1, DNA and amino acid sequence determination and analysis, sequence comparisons of isozyme KstD1-3 and other KstD enzymes, phylogenetic analysis, quantitative RT-PCR enzyme expression analysis, recombinant expression in Escherichia coli strain BL21(DE3) and Bacillus subtilis strain 6051a, subcloning in Escherichia coli strain DH5alpha, isozyme KstD1 overexpression in the DELTAkstD1 deletion mutant strain
isozyme KstD2, DNA and amino acid sequence determination and analysis, sequence comparisons of isozyme KstD1-3 and other KstD enzymes, phylogenetic analysis, quantitative RT-PCR enzyme expression analysis, recombinant expression in Escherichia coli strain BL21(DE3) and Bacillus subtilis strain 6051a, subcloning in Escherichia coli strain DH5alpha, isozyme KstD2 overexpression in the DELTAkstD1 deletion mutant strain
isozyme KstD3, DNA and amino acid sequence determination and analysis, sequence comparisons of isozyme KstD1-3 and other KstD enzymes, phylogenetic analysis, quantitative RT-PCR enzyme expression analysis, recombinant expression in Escherichia coli strain BL21(DE3) and Bacillus subtilis strain 6051a, subcloning in Escherichia coli strain DH5alpha, isozyme KstD3 overexpression in the DELTAkstD1 deletion mutant strain
recombinant expression of His-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3)
site directed mutagenesis of steroid binding site, Y-121 involved in catalytic function, Y-116, Y-104 involved in substrate binding
-
under control of the Phsp promoter and introduced into mutant cells of Mycobacterium smegmatis
-
unmarked gene deletion, biochemical evidence for a second 3-ketosteroid-DELTA1-dehydrogenase
-