BRENDA - Enzyme Database show
show all sequences of 1.14.14.84

Protein components of a cytochrome P-450 linalool 8-methyl hydroxylase

Ullah, A.H.J.; Murray, R.I.; Bhattacharyya, P.K.; Wagner, G.C.; Gunsalus, I.C.; J. Biol. Chem. 265, 1345-1351 (1990)

Data extracted from this reference:

Inhibitors
Inhibitors
Commentary
Organism
Structure
CO
complete inhibition; complete inhibition of the oxidation reactions
Pseudomonas putida
Localization
Localization
Commentary
Organism
GeneOntology No.
Textmining
cytoplasm
-
Pseudomonas putida
5737
-
Metals/Ions
Metals/Ions
Commentary
Organism
Structure
Fe2+
heme-containing cytochrome P450 enzyme; requirement of two enzyme components, iron-heme protein and iron-sulfur protein
Pseudomonas putida
Molecular Weight [Da]
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
additional information
-
multi-component enzyme consisting of LIN-reductase, Fe2-S2-redoxin and cytochrome LIN-P450. MW of the reductase: 43700 Da, analytical data from amino acid composition and prosthetic group quantification, 45000 Da, gel filtration. MW of Fe2-S2-redoxin: 11000 Da, gel filtration, 12800 Da, analytical data from amino acid composition and prosthetic group quantification. MW of cytochrome LIN-P450: 44800 Da, analytical data from amino acid composition and prosthetic group quantification, 45000 Da, gel filtration
Pseudomonas putida
43700
-
1 * 44000, SDS-PAGE, 1 * 43700, about, sequence calculation
Pseudomonas putida
44000
-
1 * 44000, SDS-PAGE, 1 * 43700, about, sequence calculation
Pseudomonas putida
45000
-
gel filtration
Pseudomonas putida
Natural Substrates/ Products (Substrates)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
3,7-dimethylocta-1,6-dien-3-ol + [reduced NADPH-hemoprotein reductase] + O2
Pseudomonas putida
linalool, initial reaction in linalool catabolism
(E)-3,7-dimethylocta-1,6-dien-3,8-diol + [oxidized NADPH-hemoprotein reductase] + H2O
-
-
?
3,7-dimethylocta-1,6-dien-3-ol + [reduced NADPH-hemoprotein reductase] + O2
Pseudomonas putida PpG777
linalool, initial reaction in linalool catabolism
(E)-3,7-dimethylocta-1,6-dien-3,8-diol + [oxidized NADPH-hemoprotein reductase] + H2O
-
-
?
linalool + 2 [reduced NADPH-hemoprotein reductase] + 2 O2
Pseudomonas putida
-
(6E)-8-oxolinalool + 2 [oxidized NADPH-hemoprotein reductase] + 3 H2O
-
-
?
linalool + 2 [reduced NADPH-hemoprotein reductase] + 2 O2
Pseudomonas putida PpG777
-
(6E)-8-oxolinalool + 2 [oxidized NADPH-hemoprotein reductase] + 3 H2O
-
-
?
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Pseudomonas putida
-
; i.e. Pseudomonas incognita
-
Pseudomonas putida PpG777
-
i.e. Pseudomonas incognita
-
Purification (Commentary)
Commentary
Organism
native enzyme by anion exchange chromatography, ammonium sulfate fractionation, a second step of anion exchange chromatography, affinity chromatography, a thrid step of anion exchange chromatography, and two further different steps of affinity chromatography to over 90% hommogeneity; purification of 3 enzyme components: iron-sulphur, FAD and P-450
Pseudomonas putida
Reaction
Reaction
Commentary
Organism
linalool + 2 [reduced NADPH-hemoprotein reductase] + 2 O2 = (6E)-8-oxolinalool + 2 [oxidized NADPH-hemoprotein reductase] + 3 H2O
mixed function monooxygenase consisting of LIN-reductase, Fe2S2-redoxin and cytochrome LIN P-450
Pseudomonas putida
Source Tissue
Source Tissue
Commentary
Organism
Textmining
additional information
strain PpG777 utilizes linalool as sole carbon and energy source
Pseudomonas putida
-
Specific Activity [micromol/min/mg]
Specific Activity Minimum [mol/min/mg]
Specific Activity Maximum [mol/min/mg]
Commentary
Organism
0.0042
-
reductase
Pseudomonas putida
0.014
-
purified enzyme, pH 7.4, temperature not specified in the publication
Pseudomonas putida
Storage Stability
Storage Stability
Organism
-196C, LIN-redoxin reductase and cytochrome LINP-450 in solution stable over a long period after ultrafiltration or dialysis and repeated freeze/thaw-cycles
Pseudomonas putida
0C, LIN-redoxin loses its prosthetic group, 5 mM DTT retards apoprotein formation
Pseudomonas putida
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
(6E)-8-hydroxylinalool + [reduced NADPH-hemoprotein reductase] + O2
-
390113
Pseudomonas putida
(6E)-8-oxolinalool + oxidized NADPH-hemoprotein reductase] + H2O
product analysis by mass spectrometry
-
-
?
(E)-3,7-dimethylocta-1,6-dien-3,8-diol + [reduced NADPH-hemoprotein reductase] + O2
8-hydroxylinalool
390113
Pseudomonas putida
8-oxolinalool + [oxidized NADPH-hemoprotein reductase] + H2O
-
390113
Pseudomonas putida
?
(E)-3,7-dimethylocta-1,6-dien-3,8-diol + [reduced NADPH-hemoprotein reductase] + O2
8-hydroxylinalool
390113
Pseudomonas putida PpG777
8-oxolinalool + [oxidized NADPH-hemoprotein reductase] + H2O
-
390113
Pseudomonas putida PpG777
?
3,7-dimethylocta-1,6-dien-3-ol + [reduced NADPH-hemoprotein reductase] + O2
-
390113
Pseudomonas putida
(E)-3,7-dimethylocta-1,6-dien-3,8-diol + [oxidized NADPH-hemoprotein reductase] + H2O
8-hydroxylinalool
390113
Pseudomonas putida
?
3,7-dimethylocta-1,6-dien-3-ol + [reduced NADPH-hemoprotein reductase] + O2
linalool, initial reaction in linalool catabolism
390113
Pseudomonas putida
(E)-3,7-dimethylocta-1,6-dien-3,8-diol + [oxidized NADPH-hemoprotein reductase] + H2O
-
-
-
?
3,7-dimethylocta-1,6-dien-3-ol + [reduced NADPH-hemoprotein reductase] + O2
-
390113
Pseudomonas putida PpG777
(E)-3,7-dimethylocta-1,6-dien-3,8-diol + [oxidized NADPH-hemoprotein reductase] + H2O
8-hydroxylinalool
390113
Pseudomonas putida PpG777
?
3,7-dimethylocta-1,6-dien-3-ol + [reduced NADPH-hemoprotein reductase] + O2
linalool, initial reaction in linalool catabolism
390113
Pseudomonas putida PpG777
(E)-3,7-dimethylocta-1,6-dien-3,8-diol + [oxidized NADPH-hemoprotein reductase] + H2O
-
-
-
?
linalool + 2 [reduced NADPH-hemoprotein reductase] + 2 O2
-
390113
Pseudomonas putida
(6E)-8-oxolinalool + 2 [oxidized NADPH-hemoprotein reductase] + 3 H2O
-
-
-
?
linalool + 2 [reduced NADPH-hemoprotein reductase] + 2 O2
overall reaction
390113
Pseudomonas putida
(6E)-8-oxolinalool + 2 [oxidized NADPH-hemoprotein reductase] + 3 H2O
product analysis by mass spectrometry
-
-
?
linalool + 2 [reduced NADPH-hemoprotein reductase] + 2 O2
-
390113
Pseudomonas putida PpG777
(6E)-8-oxolinalool + 2 [oxidized NADPH-hemoprotein reductase] + 3 H2O
-
-
-
?
linalool + [reduced NADPH-hemoprotein reductase] + O2
-
390113
Pseudomonas putida
(6E)-8-hydroxylinalool + [oxidized NADPH-hemoprotein reductase] + H2O
product analysis by mass spectrometry
-
-
?
additional information
the enzyme does not perform 10-methyl hydroxylation of linalool
390113
Pseudomonas putida
?
-
-
-
-
additional information
the enzyme does not perform 10-methyl hydroxylation of linalool
390113
Pseudomonas putida PpG777
?
-
-
-
-
Subunits
Subunits
Commentary
Organism
monomer
1 * 44000, SDS-PAGE, 1 * 43700, about, sequence calculation
Pseudomonas putida
More
enzyme consists of LIN-reductase, MW 44000 Da, and Fe2S2-redoxin, MW 10700 Da, and cytochrome LIN-P450, MW 47000 Da, SDS-PAGE
Pseudomonas putida
Turnover Number [1/s]
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
2
-
NADH
substrate-stimulated NADH oxidation using excess amounts of P-450 and FAD component of the enzyme
Pseudomonas putida
32
-
NADH
substrate-stimulated NADH oxidation using excess amounts of iron-sulfur and FAD component of the enzyme
Pseudomonas putida
43
-
NADH
substrate-stimulated NADH oxidation using excess amounts of iron sulfur and P-450 component of the enzyme
Pseudomonas putida
pH Optimum
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
7.4
-
assay at
Pseudomonas putida
Cofactor
Cofactor
Commentary
Organism
Structure
cytochrome P450
-
Pseudomonas putida
heme
-
Pseudomonas putida
redoxin
an Fe2/S2/Cys4 protein
Pseudomonas putida
pI Value
Organism
Commentary
pI Value Maximum
pI Value
Pseudomonas putida
sequence calculation
-
4.65
Cofactor (protein specific)
Cofactor
Commentary
Organism
Structure
cytochrome P450
-
Pseudomonas putida
heme
-
Pseudomonas putida
redoxin
an Fe2/S2/Cys4 protein
Pseudomonas putida
Inhibitors (protein specific)
Inhibitors
Commentary
Organism
Structure
CO
complete inhibition; complete inhibition of the oxidation reactions
Pseudomonas putida
Localization (protein specific)
Localization
Commentary
Organism
GeneOntology No.
Textmining
cytoplasm
-
Pseudomonas putida
5737
-
Metals/Ions (protein specific)
Metals/Ions
Commentary
Organism
Structure
Fe2+
heme-containing cytochrome P450 enzyme; requirement of two enzyme components, iron-heme protein and iron-sulfur protein
Pseudomonas putida
Molecular Weight [Da] (protein specific)
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
additional information
-
multi-component enzyme consisting of LIN-reductase, Fe2-S2-redoxin and cytochrome LIN-P450. MW of the reductase: 43700 Da, analytical data from amino acid composition and prosthetic group quantification, 45000 Da, gel filtration. MW of Fe2-S2-redoxin: 11000 Da, gel filtration, 12800 Da, analytical data from amino acid composition and prosthetic group quantification. MW of cytochrome LIN-P450: 44800 Da, analytical data from amino acid composition and prosthetic group quantification, 45000 Da, gel filtration
Pseudomonas putida
43700
-
1 * 44000, SDS-PAGE, 1 * 43700, about, sequence calculation
Pseudomonas putida
44000
-
1 * 44000, SDS-PAGE, 1 * 43700, about, sequence calculation
Pseudomonas putida
45000
-
gel filtration
Pseudomonas putida
Natural Substrates/ Products (Substrates) (protein specific)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
3,7-dimethylocta-1,6-dien-3-ol + [reduced NADPH-hemoprotein reductase] + O2
Pseudomonas putida
linalool, initial reaction in linalool catabolism
(E)-3,7-dimethylocta-1,6-dien-3,8-diol + [oxidized NADPH-hemoprotein reductase] + H2O
-
-
?
3,7-dimethylocta-1,6-dien-3-ol + [reduced NADPH-hemoprotein reductase] + O2
Pseudomonas putida PpG777
linalool, initial reaction in linalool catabolism
(E)-3,7-dimethylocta-1,6-dien-3,8-diol + [oxidized NADPH-hemoprotein reductase] + H2O
-
-
?
linalool + 2 [reduced NADPH-hemoprotein reductase] + 2 O2
Pseudomonas putida
-
(6E)-8-oxolinalool + 2 [oxidized NADPH-hemoprotein reductase] + 3 H2O
-
-
?
linalool + 2 [reduced NADPH-hemoprotein reductase] + 2 O2
Pseudomonas putida PpG777
-
(6E)-8-oxolinalool + 2 [oxidized NADPH-hemoprotein reductase] + 3 H2O
-
-
?
Purification (Commentary) (protein specific)
Commentary
Organism
native enzyme by anion exchange chromatography, ammonium sulfate fractionation, a second step of anion exchange chromatography, affinity chromatography, a thrid step of anion exchange chromatography, and two further different steps of affinity chromatography to over 90% hommogeneity; purification of 3 enzyme components: iron-sulphur, FAD and P-450
Pseudomonas putida
Source Tissue (protein specific)
Source Tissue
Commentary
Organism
Textmining
additional information
strain PpG777 utilizes linalool as sole carbon and energy source
Pseudomonas putida
-
Specific Activity [micromol/min/mg] (protein specific)
Specific Activity Minimum [mol/min/mg]
Specific Activity Maximum [mol/min/mg]
Commentary
Organism
0.0042
-
reductase
Pseudomonas putida
0.014
-
purified enzyme, pH 7.4, temperature not specified in the publication
Pseudomonas putida
Storage Stability (protein specific)
Storage Stability
Organism
-196C, LIN-redoxin reductase and cytochrome LINP-450 in solution stable over a long period after ultrafiltration or dialysis and repeated freeze/thaw-cycles
Pseudomonas putida
0C, LIN-redoxin loses its prosthetic group, 5 mM DTT retards apoprotein formation
Pseudomonas putida
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
(6E)-8-hydroxylinalool + [reduced NADPH-hemoprotein reductase] + O2
-
390113
Pseudomonas putida
(6E)-8-oxolinalool + oxidized NADPH-hemoprotein reductase] + H2O
product analysis by mass spectrometry
-
-
?
(E)-3,7-dimethylocta-1,6-dien-3,8-diol + [reduced NADPH-hemoprotein reductase] + O2
8-hydroxylinalool
390113
Pseudomonas putida
8-oxolinalool + [oxidized NADPH-hemoprotein reductase] + H2O
-
390113
Pseudomonas putida
?
(E)-3,7-dimethylocta-1,6-dien-3,8-diol + [reduced NADPH-hemoprotein reductase] + O2
8-hydroxylinalool
390113
Pseudomonas putida PpG777
8-oxolinalool + [oxidized NADPH-hemoprotein reductase] + H2O
-
390113
Pseudomonas putida PpG777
?
3,7-dimethylocta-1,6-dien-3-ol + [reduced NADPH-hemoprotein reductase] + O2
-
390113
Pseudomonas putida
(E)-3,7-dimethylocta-1,6-dien-3,8-diol + [oxidized NADPH-hemoprotein reductase] + H2O
8-hydroxylinalool
390113
Pseudomonas putida
?
3,7-dimethylocta-1,6-dien-3-ol + [reduced NADPH-hemoprotein reductase] + O2
linalool, initial reaction in linalool catabolism
390113
Pseudomonas putida
(E)-3,7-dimethylocta-1,6-dien-3,8-diol + [oxidized NADPH-hemoprotein reductase] + H2O
-
-
-
?
3,7-dimethylocta-1,6-dien-3-ol + [reduced NADPH-hemoprotein reductase] + O2
-
390113
Pseudomonas putida PpG777
(E)-3,7-dimethylocta-1,6-dien-3,8-diol + [oxidized NADPH-hemoprotein reductase] + H2O
8-hydroxylinalool
390113
Pseudomonas putida PpG777
?
3,7-dimethylocta-1,6-dien-3-ol + [reduced NADPH-hemoprotein reductase] + O2
linalool, initial reaction in linalool catabolism
390113
Pseudomonas putida PpG777
(E)-3,7-dimethylocta-1,6-dien-3,8-diol + [oxidized NADPH-hemoprotein reductase] + H2O
-
-
-
?
linalool + 2 [reduced NADPH-hemoprotein reductase] + 2 O2
-
390113
Pseudomonas putida
(6E)-8-oxolinalool + 2 [oxidized NADPH-hemoprotein reductase] + 3 H2O
-
-
-
?
linalool + 2 [reduced NADPH-hemoprotein reductase] + 2 O2
overall reaction
390113
Pseudomonas putida
(6E)-8-oxolinalool + 2 [oxidized NADPH-hemoprotein reductase] + 3 H2O
product analysis by mass spectrometry
-
-
?
linalool + 2 [reduced NADPH-hemoprotein reductase] + 2 O2
-
390113
Pseudomonas putida PpG777
(6E)-8-oxolinalool + 2 [oxidized NADPH-hemoprotein reductase] + 3 H2O
-
-
-
?
linalool + [reduced NADPH-hemoprotein reductase] + O2
-
390113
Pseudomonas putida
(6E)-8-hydroxylinalool + [oxidized NADPH-hemoprotein reductase] + H2O
product analysis by mass spectrometry
-
-
?
additional information
the enzyme does not perform 10-methyl hydroxylation of linalool
390113
Pseudomonas putida
?
-
-
-
-
additional information
the enzyme does not perform 10-methyl hydroxylation of linalool
390113
Pseudomonas putida PpG777
?
-
-
-
-
Subunits (protein specific)
Subunits
Commentary
Organism
monomer
1 * 44000, SDS-PAGE, 1 * 43700, about, sequence calculation
Pseudomonas putida
More
enzyme consists of LIN-reductase, MW 44000 Da, and Fe2S2-redoxin, MW 10700 Da, and cytochrome LIN-P450, MW 47000 Da, SDS-PAGE
Pseudomonas putida
Turnover Number [1/s] (protein specific)
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
2
-
NADH
substrate-stimulated NADH oxidation using excess amounts of P-450 and FAD component of the enzyme
Pseudomonas putida
32
-
NADH
substrate-stimulated NADH oxidation using excess amounts of iron-sulfur and FAD component of the enzyme
Pseudomonas putida
43
-
NADH
substrate-stimulated NADH oxidation using excess amounts of iron sulfur and P-450 component of the enzyme
Pseudomonas putida
pH Optimum (protein specific)
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
7.4
-
assay at
Pseudomonas putida
pI Value (protein specific)
Organism
Commentary
pI Value Maximum
pI Value
Pseudomonas putida
sequence calculation
-
4.65
General Information
General Information
Commentary
Organism
metabolism
the enzyme catalyzes the first step in linalool catabolism
Pseudomonas putida
additional information
analysis of the heme-thiolate reaction center, molecular domains of substrate specificity, redox-pair interactions, and the regulation of the reaction cycle, overview
Pseudomonas putida
General Information (protein specific)
General Information
Commentary
Organism
metabolism
the enzyme catalyzes the first step in linalool catabolism
Pseudomonas putida
additional information
analysis of the heme-thiolate reaction center, molecular domains of substrate specificity, redox-pair interactions, and the regulation of the reaction cycle, overview
Pseudomonas putida
Other publictions for EC 1.14.14.84
No.
1st author
Pub Med
title
organims
journal
volume
pages
year
Activating Compound
Application
Cloned(Commentary)
Crystallization (Commentary)
Engineering
General Stability
Inhibitors
KM Value [mM]
Localization
Metals/Ions
Molecular Weight [Da]
Natural Substrates/ Products (Substrates)
Organic Solvent Stability
Organism
Oxidation Stability
Posttranslational Modification
Purification (Commentary)
Reaction
Renatured (Commentary)
Source Tissue
Specific Activity [micromol/min/mg]
Storage Stability
Substrates and Products (Substrate)
Subunits
Temperature Optimum [C]
Temperature Range [C]
Temperature Stability [C]
Turnover Number [1/s]
pH Optimum
pH Range
pH Stability
Cofactor
Ki Value [mM]
pI Value
IC50 Value
Activating Compound (protein specific)
Application (protein specific)
Cloned(Commentary) (protein specific)
Cofactor (protein specific)
Crystallization (Commentary) (protein specific)
Engineering (protein specific)
General Stability (protein specific)
IC50 Value (protein specific)
Inhibitors (protein specific)
Ki Value [mM] (protein specific)
KM Value [mM] (protein specific)
Localization (protein specific)
Metals/Ions (protein specific)
Molecular Weight [Da] (protein specific)
Natural Substrates/ Products (Substrates) (protein specific)
Organic Solvent Stability (protein specific)
Oxidation Stability (protein specific)
Posttranslational Modification (protein specific)
Purification (Commentary) (protein specific)
Renatured (Commentary) (protein specific)
Source Tissue (protein specific)
Specific Activity [micromol/min/mg] (protein specific)
Storage Stability (protein specific)
Substrates and Products (Substrate) (protein specific)
Subunits (protein specific)
Temperature Optimum [C] (protein specific)
Temperature Range [C] (protein specific)
Temperature Stability [C] (protein specific)
Turnover Number [1/s] (protein specific)
pH Optimum (protein specific)
pH Range (protein specific)
pH Stability (protein specific)
pI Value (protein specific)
Expression
General Information
General Information (protein specific)
Expression (protein specific)
KCat/KM [mM/s]
KCat/KM [mM/s] (protein specific)
701873
Bell
A cytochrome P450 class I elec ...
Novosphingobium aromaticivorans, Novosphingobium aromaticivorans ATCC 700278D-5
Appl. Microbiol. Biotechnol.
86
163-175
2010
-
-
1
-
1
-
-
-
-
1
-
2
-
2
-
-
1
-
-
-
-
-
3
-
1
-
-
-
1
-
-
2
-
-
-
-
-
1
2
-
1
-
-
-
-
-
-
1
-
2
-
-
-
1
-
-
-
-
3
-
1
-
-
-
1
-
-
-
-
1
1
-
-
-
717712
Ropp
Cloning and expression of a me ...
Pseudomonas putida, Pseudomonas putida PpG777
J. Bacteriol.
175
6028-6037
1993
-
-
1
-
-
-
-
-
-
1
-
2
-
2
-
-
1
-
-
-
-
-
2
-
-
-
-
-
-
-
-
2
-
-
-
-
-
1
2
-
-
-
-
-
-
-
-
1
-
2
-
-
-
1
-
-
-
-
2
-
-
-
-
-
-
-
-
-
-
3
3
-
-
-
390114
Bernhardt
Reconstitution of cytochrome P ...
Pseudomonas putida
Biochem. Biophys. Res. Commun.
187
310-317
1992
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
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-
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-
-
-
-
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-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
390113
Ullah
Protein components of a cytoch ...
Pseudomonas putida, Pseudomonas putida PpG777
J. Biol. Chem.
265
1345-1351
1990
-
-
-
-
-
-
1
-
1
1
4
4
-
3
-
-
1
1
-
1
2
2
13
2
-
-
-
3
1
-
-
3
-
1
-
-
-
-
3
-
-
-
-
1
-
-
1
1
4
4
-
-
-
1
-
1
2
2
13
2
-
-
-
3
1
-
-
1
-
2
2
-
-
-
390112
Bhattacharyya
-
Chemical probes into the activ ...
Pseudomonas putida, Pseudomonas putida PpG777
Proc. Indian Acad. Sci. Chem. Sci.
93
1289-1304
1984
-
-
-
-
-
-
-
-
-
-
-
-
-
3
-
-
-
-
-
-
-
-
6
-
1
-
-
1
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
6
-
1
-
-
1
1
-
-
-
-
-
-
-
-
-