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Literature summary extracted from

  • Florin, C.; Koehler, T.; Grandguillot, M.; Plesiat, P.
    Comamonas testosteroni 3-ketosteroid-DELTA4(5alpha)-dehydrogenase (1996), J. Bacteriol., 178, 3322-3330.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.3.99.5 expression in Escherichia coli JM109 carrying pXE25, activity is maximal 18times higher than the activity expressed by induced cells of Comamonas testosteroni. Overexpression in Pseudomonas putida KT2440 using pMMB208 as expression vector, activity is maximaly 264times higher than the activity expressed by induced cells of Comamonas testosteroni Comamonas testosteroni

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.3.99.5 0.0066
-
 1-Androstene-3,17-dione reaction with N,N,N‘,N‘-tetramethyl-1,4-phenylenediamine Comamonas testosteroni
1.3.99.5 0.0161
-
 5alpha-androstane-3,17-dione reaction with N,N,N‘,N‘-tetramethyl-1,4-phenylenediamine Comamonas testosteroni

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.3.99.5 58100
-
 x * 58100, calculation from nucleotide sequence Comamonas testosteroni
1.3.99.5 60000
-
 x * 60000, SDS-PAGE Comamonas testosteroni

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.3.99.5 testosterone + reduced acceptor Comamonas testosteroni key enzyme in the degradation of steroids having an A:B ring fusion in a trans configuration 5alpha-dihydrotestosterone + acceptor
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
1.3.99.5 Comamonas testosteroni
-
 formerly Pseudomonas testosteroni
-

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.3.99.5 1-androstene-3,17-dione + phenazine methosulfate acceptor: N,N,N‘,N‘-tetramethyl-1,4-phenylenediamine, menadione, phenazine methosulfate, cytochrome c Comamonas testosteroni 1,4-androstadiene-3,17-dione + reduced phenazine methosulfate
-
 ir
1.3.99.5 5alpha-1-androsten-3,17-dione + acceptor acceptor: N,N,N‘,N‘-tetramethyl-1,4-phenylenediamine, menadione, phenazine methosulfate, cytochrome c Comamonas testosteroni ? + reduced acceptor
-
 ir
1.3.99.5 5alpha-androstane-3,17-dione + N,N,N',N'-tetramethyl-1,4-phenylendiamine
-
 Comamonas testosteroni androst-4-ene-3,17-dione + reduced N,N,N',N'-tetramethyl-1,4-phenylendiamine
-
 r
1.3.99.5 testosterone + reduced acceptor key enzyme in the degradation of steroids having an A:B ring fusion in a trans configuration Comamonas testosteroni 5alpha-dihydrotestosterone + acceptor
-
 ?

Subunits

EC Number Subunits Comment Organism
1.3.99.5 ? x * 60000, SDS-PAGE Comamonas testosteroni
1.3.99.5 ? x * 58100, calculation from nucleotide sequence Comamonas testosteroni

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.3.99.5 40
-
 reaction with 1-androstene-3,17-dione and N,N,N‘,N‘-tetramethyl-1,4-phenylenediamine Comamonas testosteroni

Temperature Range [°C]

EC Number Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
1.3.99.5 25 50 25°C: about 50% of maximal activity, 50°C: about 20% of maximal activity, reaction with N,N,N‘,N‘-tetramethyl-1,4-phenylenediamine Comamonas testosteroni

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.3.99.5 8 9 reaction with 1-androstene-3,17-dione and N,N,N‘,N‘-tetramethyl-1,4-phenylenediamine Comamonas testosteroni

pH Range

EC Number pH Minimum pH Maximum Comment Organism
1.3.99.5 7 9 pH 7.0: about 40% of maximal activity, pH 8.0-9.0: maximal activity, reaction with Delta1-androstene-3,17-dione and N,N,N‘,N‘-tetramethyl-1,4-phenylenediamine Comamonas testosteroni