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Literature summary extracted from

  • Alonso, H.; Roujeinikova, A.
    Characterization and two-dimensional crystallization of membrane component AlkB of the medium-chain alkane hydroxylase system from Pseudomonas putida GPo1 (2012), Appl. Environ. Microbiol., 78, 7946-7953.
    View publication on PubMedView publication on EuropePMC

Application

EC Number Application Comment Organism
1.14.15.3 synthesis bacterial alkane hydroxylases have tremendous potential as biocatalysts for the stereo- and regioselective transformation of a wide range of chemically inert unreactive alkanes into valuable reactive chemical precursors Pseudomonas putida

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.14.15.3 gene alkB, expression of Strep-tagged enzyme in Escherichia coli strains BL21(DE3), C41(DE3), Rosetta-2(DE3), pLysS, and BL21-CondonPlus(DE3)-RIPL under different growth conditions Pseudomonas putida

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
1.14.15.3 2D crystallization Pseudomonas putida

Inhibitors

EC Number Inhibitors Comment Organism Structure
1.14.15.3 1-octyne acts as a mechanism-based inhibitor of AlkB Pseudomonas putida
1.14.15.3 n-decyl-beta-D-maltopyranoside
-
 Pseudomonas putida
1.14.15.3 n-dodecyl-beta-D-maltopyranoside
-
 Pseudomonas putida
1.14.15.3 n-dodecyl-N,N-dimethylamine-n-oxide
-
 Pseudomonas putida
1.14.15.3 octyl glucose neopentyl glycol
-
 Pseudomonas putida

Localization

EC Number Localization Comment Organism GeneOntology No. Textmining
1.14.15.3 membrane AlkB is the integral membrane component of the Pseudomonas putida alkane hydroxylase system Pseudomonas putida 16020
-

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.14.15.3 46000
-
 x * 46000, recombinant Strep-tagged AlkB, SDS-PAGE Pseudomonas putida

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.14.15.3 octane + reduced rubredoxin + O2 Pseudomonas putida
-
 1-octanol + oxidized rubredoxin + H2O
-
 ?
1.14.15.3 octane + reduced rubredoxin + O2 Pseudomonas putida GPo1 / ATCC29347
-
 1-octanol + oxidized rubredoxin + H2O
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
1.14.15.3 Pseudomonas putida
-
-
-
1.14.15.3 Pseudomonas putida GPo1 / ATCC29347
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.14.15.3 recombinant Strep-tagged enzyme from Escherichia coli strains BL21(DE3), C41(DE3), Rosetta-2(DE3), pLysS, and BL21-CondonPlus(DE3)-RIPL by Strep-tag affinity chromatography, ultrafiltration, and gel filtration Pseudomonas putida

Source Tissue

EC Number Source Tissue Comment Organism Textmining
1.14.15.3 additional information the alkane hydroxylase system of Pseudomonas putida GPo1 allows it to use alkanes as the sole source of carbon and energy Pseudomonas putida
-

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.14.15.3 octane + reduced rubredoxin + O2
-
 Pseudomonas putida 1-octanol + oxidized rubredoxin + H2O
-
 ?
1.14.15.3 octane + reduced rubredoxin + O2
-
 Pseudomonas putida GPo1 / ATCC29347 1-octanol + oxidized rubredoxin + H2O
-
 ?

Subunits

EC Number Subunits Comment Organism
1.14.15.3 ? x * 46000, recombinant Strep-tagged AlkB, SDS-PAGE Pseudomonas putida
1.14.15.3 More the physiologically active form of membrane-embedded AlkB may be a multimer, the monomeric form of AlkB produced by purification in n-decyl-beta-D-maltopyranoside, n-dodecyl-beta-D-maltopyranoside, octyl glucose neopentyl glycol, and n-dodecyl-N,N-dimethylamine-N-oxide is largely inactive. In contrast, detergents that do not disrupt the AlkB oligomeric state, i.e. decyl maltose neopentyl glycol, lauryl maltose neopentyl glycol, and octaethylene glycol monododecyl ether, preserve the enzyme activity at a level close to that of the detergent-free control sample Pseudomonas putida
1.14.15.3 trimer AlkB reconstituted into a lipid bilayer forms trimers Pseudomonas putida

Synonyms

EC Number Synonyms Comment Organism
1.14.15.3 AlkB
-
 Pseudomonas putida
1.14.15.3 medium-chain alkane hydroxylase
-
 Pseudomonas putida

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.14.15.3 22
-
 assay at room temperature Pseudomonas putida

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
1.14.15.3 65
-
 purified recombinant enzyme, 15 min, inactivation Pseudomonas putida

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.14.15.3 8
-
 assay at Pseudomonas putida

Cofactor

EC Number Cofactor Comment Organism Structure
1.14.15.3 rubredoxin
-
 Pseudomonas putida

General Information

EC Number General Information Comment Organism
1.14.15.3 physiological function the alkane hydroxylase system of Pseudomonas putida GPo1 allows it to use alkanes as the sole source of carbon and energy. The alkane hydroxylase system of Pseudomonas putida GPo1 comprises three protein components: AlkB, soluble NADH-rubredoxin reductase, and soluble electron transfer protein rubredoxin. AlkB transfers one oxygen atom from O2 to the alkane molecule, while the other oxygen is reduced to H2O using the electrons provided by NADH-rubredoxin reductase via rubredoxin Pseudomonas putida