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Literature summary for 1.1.1.218 extracted from

  • Bruce, N.C.; Wilmot, C.J.; Jordan, K.N.; Stephens, L.D.G.; Lowe, Ch.R.
    Microbial degradation of the morphine alkaloids (1991), Biochem. J., 274, 875-880.
No PubMed abstract available

General Stability

General Stability Organism
2-mercaptoethanol stabilizes Pseudomonas putida
dithiothreitol stabilizes Pseudomonas putida

Inhibitors

Inhibitors Comment Organism Structure
1,10-phenanthroline
-
Pseudomonas putida
2,2'-dipyridyl
-
Pseudomonas putida
CuSO4
-
Pseudomonas putida
iodoacetate
-
Pseudomonas putida
additional information no inhibition by EDTA, NEM, hydroxyquinoline Pseudomonas putida
NEM inhibit activity at much higher concentration (e.g. at 1 mM) Pseudomonas putida

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.044
-
codeine with NADP+ Pseudomonas putida
0.35
-
NADP+
-
Pseudomonas putida
0.46
-
morphine cosubstrate NADP+ Pseudomonas putida
2.91
-
Dihydrocodeine cosubstrate NADP+ Pseudomonas putida

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
31000
-
denatured enzyme, SDS-PAGE Pseudomonas putida
32000
-
native enzyme, gel filtration Pseudomonas putida
310000
-
1 * 310000, SDS-PAGE Pseudomonas putida

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
morphine + NADP+ Pseudomonas putida highly specific alkaloid dehydrogenase, oxidizing only the C-6 hydroxy group of morphine and codeine, morphine as the sole carbon and energy source morphinone + NADPH
-
?
morphine + NADP+ Pseudomonas putida M10 highly specific alkaloid dehydrogenase, oxidizing only the C-6 hydroxy group of morphine and codeine, morphine as the sole carbon and energy source morphinone + NADPH
-
?

Organism

Organism UniProt Comment Textmining
Pseudomonas putida
-
M10
-
Pseudomonas putida M10
-
M10
-

Purification (Commentary)

Purification (Comment) Organism
gel filtration, various chromatographic techniques Pseudomonas putida

Specific Activity [micromol/min/mg]

Specific Activity Minimum [┬Ámol/min/mg] Specific Activity Maximum [┬Ámol/min/mg] Comment Organism
0.08
-
morphine, NADP+ Pseudomonas putida

Storage Stability

Storage Stability Organism
-80┬░C, 10% loss of activity over 2 months Pseudomonas putida

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
codeine + NAD(P)+ oxidation at 120% the rate of morphine oxidation Pseudomonas putida codeinone + NAD(P)H at pH 6.5, + NADPH, reduction of codeinone to codeine r
codeine + NAD(P)+ oxidation at 120% the rate of morphine oxidation Pseudomonas putida M10 codeinone + NAD(P)H at pH 6.5, + NADPH, reduction of codeinone to codeine r
dihydrocodeine + NADP+
-
Pseudomonas putida dihydrocodeinone + NADPH
-
?
dihydrocodeine + NADP+
-
Pseudomonas putida M10 dihydrocodeinone + NADPH
-
?
morphine + NADP+ enzyme activity only with NADP+ Pseudomonas putida morphinone + NADPH + H+
-
?
morphine + NADP+ enzyme activity only with NADP+ Pseudomonas putida M10 morphinone + NADPH + H+
-
?
morphine + NADP+ highly specific alkaloid dehydrogenase, oxidizing only the C-6 hydroxy group of morphine and codeine, morphine as the sole carbon and energy source Pseudomonas putida morphinone + NADPH
-
?
morphine + NADP+ highly specific alkaloid dehydrogenase, oxidizing only the C-6 hydroxy group of morphine and codeine, morphine as the sole carbon and energy source Pseudomonas putida M10 morphinone + NADPH
-
?

Subunits

Subunits Comment Organism
monomer 1 * 310000, SDS-PAGE Pseudomonas putida

Temperature Optimum [┬░C]

Temperature Optimum [┬░C] Temperature Optimum Maximum [┬░C] Comment Organism
30
-
enzyme assay Pseudomonas putida

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
6.5
-
in presence of NADPH reduction of codeinone to codeine, in MOPS buffer Pseudomonas putida
9.5
-
with NADP+ as cofactor, in glycine-NaOH buffer, oxidizing the C-6 hydroxy group of morphine and codeine Pseudomonas putida

Cofactor

Cofactor Comment Organism Structure
NADP+ the Pseudomonas enzyme acts only with NADP+ Pseudomonas putida