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Literature summary extracted from

  • Wei, W.; Wang, F.Q.; Fan, S.Y.; Wei, D.Z.
    Inactivation and augmentation of the primary 3-ketosteroid-DELTA1-dehydrogenase in Mycobacterium neoaurum NwIB-01: biotransformation of soybean phytosterols to 4-androstene-3,17-dione or 1,4-androstadiene-3,17-dione (2010), Appl. Environ. Microbiol., 76, 4578-4582.
    View publication on PubMedView publication on EuropePMC

Application

EC Number Application Comment Organism
1.3.99.4 synthesis engineered enzyme mutants, in which KsdDM is inactivated or augmented, are useful for production of phytosterols 4-androstene-3,17-dione or 1,4-androstadiene-3,17-dione, circumventing the difficulty of separating 4-androstene-3,17-dione from 1,4-androstadiene-3,17-dione, a key bottleneck to the microbial transformation of phytosterols in industry Mycolicibacterium neoaurum

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.3.99.4 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 Mycolicibacterium neoaurum

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.3.99.4 4-androstene-3,17-dione + FAD Mycolicibacterium neoaurum
-
 1,4-androstadiene-3,17-dione + FADH2
-
 ?
1.3.99.4 4-androstene-3,17-dione + FAD Mycolicibacterium neoaurum NwIB-01
-
 1,4-androstadiene-3,17-dione + FADH2
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
1.3.99.4 Mycolicibacterium neoaurum C6KID9
-
-
1.3.99.4 Mycolicibacterium neoaurum NwIB-01 C6KID9
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.3.99.4 recombinant His-tagged enzyme by nickel affinity chromatography Mycolicibacterium neoaurum

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
1.3.99.4 0.8
-
 soluble recombinant His-tagged enzyme, cell extract, pH 7.0, 30°C Mycolicibacterium neoaurum

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.3.99.4 4-androstene-3,17-dione + FAD
-
 Mycolicibacterium neoaurum 1,4-androstadiene-3,17-dione + FADH2
-
 ?
1.3.99.4 4-androstene-3,17-dione + FAD with artificial electron acceptors phenazine methosulfate and dichlorophenolindophenol Mycolicibacterium neoaurum 1,4-androstadiene-3,17-dione + FADH2
-
 ?
1.3.99.4 4-androstene-3,17-dione + FAD
-
 Mycolicibacterium neoaurum NwIB-01 1,4-androstadiene-3,17-dione + FADH2
-
 ?
1.3.99.4 4-androstene-3,17-dione + FAD with artificial electron acceptors phenazine methosulfate and dichlorophenolindophenol Mycolicibacterium neoaurum NwIB-01 1,4-androstadiene-3,17-dione + FADH2
-
 ?
1.3.99.4 9alpha-hydroxy-4-androstene-3,17-dione + FAD
-
 Mycolicibacterium neoaurum 9alpha-hydroxy-1,4-androstadiene-3,17-dione + FADH2
-
 ?
1.3.99.4 9alpha-hydroxy-4-androstene-3,17-dione + FAD
-
 Mycolicibacterium neoaurum NwIB-01 9alpha-hydroxy-1,4-androstadiene-3,17-dione + FADH2
-
 ?

Synonyms

EC Number Synonyms Comment Organism
1.3.99.4 3-ketosteroid-DELTA1-dehydrogenase
-
 Mycolicibacterium neoaurum
1.3.99.4 KsdDM
-
 Mycolicibacterium neoaurum
1.3.99.4 steroid-1-dehydrogenase
-
 Mycolicibacterium neoaurum

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.3.99.4 30
-
 assay at Mycolicibacterium neoaurum

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.3.99.4 7
-
 assay at Mycolicibacterium neoaurum

Cofactor

EC Number Cofactor Comment Organism Structure
1.3.99.4 FAD putative N-terminal FAD-binding motif in KsdDM is GSG(A/G)(A/G)(A/G)X17E Mycolicibacterium neoaurum