Substrate specificity of the alkane hydroxylase system of Pseudomonas oleovorans GPo1

Beilen, Jan B. van; Kingma, Jaap; Witholt, Bernard

doi:10.1016/0141-0229(94)90066-3

Cited by 126 publications

(116 citation statements)

References 14 publications

Supporting

Mentioning

108

Contrasting

Order By: Relevance

“…The alkB hydroxylase from Pseudomonas putida GPo1 is capable of oxidizing large (.C 12 ) linear alkanes and substituted cyclic alkanes at similar rates (van Beilen et al, 1994), while also oxidizing the smaller growth substrates propane and butane with high affinity (K s 66 and 13 mM, respectively) (Johnson & Hyman, 2006). The alkane hydroxylase from propaneutilizing Mycobacterium vaccae JOB5 has also recently been characterized as an alkB hydroxylase (Lopes Ferreira et al, 2007), and has a low reported K s for propane of 3.3-4.4 mM but high K s values for branched substrates tertbutyl alcohol and methyl tert-butyl ether (1.36 and 1.18 mM, respectively) .…”

Section: Substrate Specificitymentioning

confidence: 99%

Kinetic characterization of the soluble butane monooxygenase from Thauera butanivorans, formerly ‘Pseudomonas butanovora’

et al. 2009

View full text Add to dashboard Cite

Soluble butane monooxygenase (sBMO), a three-component di-iron monooxygenase complex expressed by the C 2 -C 9 alkane-utilizing bacterium Thauera butanivorans, was kinetically characterized by measuring substrate specificities for C 1 -C 5 alkanes and product inhibition profiles. sBMO has high sequence homology with soluble methane monooxygenase (sMMO) and shares a similar substrate range, including gaseous and liquid alkanes, aromatics, alkenes and halogenated xenobiotics. Results indicated that butane was the preferred substrate (defined by k cat : K m ratios). Relative rates of oxidation for C 1 -C 5 alkanes differed minimally, implying that substrate specificity is heavily influenced by differences in substrate K m values. The low micromolar K m for linear C 2 -C 5 alkanes and the millimolar K m for methane demonstrate that sBMO is two to three orders of magnitude more specific for physiologically relevant substrates of T. butanivorans. Methanol, the product of methane oxidation and also a substrate itself, was found to have similar K m and k cat values to those of methane. This inability to kinetically discriminate between the C 1 alkane and C 1 alcohol is observed as a steady-state concentration of methanol during the two-step oxidation of methane to formaldehyde by sBMO. Unlike methanol, alcohols with chain length C 2 -C 5 do not compete effectively with their respective alkane substrates. Results from product inhibition experiments suggest that the geometry of the active site is optimized for linear molecules four to five carbons in length and is influenced by the regulatory protein component B (butane monooxygenase regulatory component; BMOB). The data suggest that alkane oxidation by sBMO is highly specialized for the turnover of C 3 -C 5 alkanes and the release of their respective alcohol products. Additionally, sBMO is particularly efficient at preventing methane oxidation during growth on linear alkanes ¢C 2, despite its high sequence homology with sMMO. These results represent, to the best of our knowledge, the first kinetic in vitro characterization of the closest known homologue of sMMO.

show abstract

Section: Substrate Specificitymentioning

confidence: 99%

Kinetic characterization of the soluble butane monooxygenase from Thauera butanivorans, formerly ‘Pseudomonas butanovora’

et al. 2009

View full text Add to dashboard Cite

show abstract

“…Recently, enzyme systems related to the P. putida GPo1 alkane hydroxylase were cloned from bacterial strains belonging to several different genera [34,50,[52][53][54]. While the P. putida GPo1 system acts on alkanes ranging from pentane to dodecane (C 5 -C 12 ) [42,55], most related membrane-bound alkane hydroxylases solely hydroxylate alkanes containing more than 10 carbon atoms [34] (see below). Several cytochrome P450 isozymes involved in alkane assimilation were recently cloned from the yeasts Candida maltosa, Yarrowia lipolytica and Debaryomyces hansenii [29,56,57].…”

Section: Streptomycesmentioning

confidence: 99%

“…P. putida GPo1 was found to oxidize many other substrates as well (including ethylsubstituted heterocyclic compounds, allyl phenyl or allyl benzyl ethers, thioethers, branched alkanes, etc.) with excellent regio-and/or stereoselectivity [16,55,[108][109][110][111]. Screening of a collection of alkane-degrading strains for the regio-and stereoselective hydroxylation of N-benzylpyrrolidine showed that 25-30% of the strains oxidized this substrate to N-benzyl-3-hydroxypyrrolidine with varying enantiomeric excess [110].…”

Section: Applications Of Alkane Hydroxylase Systemsmentioning

confidence: 99%

Diversity of Alkane Hydroxylase Systems in the Environment

Beilen

Duetz

et al. 2003

Oil & Gas Science and Technology - Rev. IFP

320

227

View full text Add to dashboard Cite

Résumé -Diversité des systèmes alcane hydroxylase dans l'environnement -La première étape dans la dégradation aérobie des alcanes par les bactéries, les levures et les champignons est catalysée par des oxygénases, une classe d'enzymes capables d'introduire des atomes d'oxygène issus de l'oxygène moléculaire dans le substrat alcane. Ces enzymes jouent un rôle important dans la biodégradation du pétrole et dans la biodégradation cométabolique de composés tels que le trichloroéthylène et les éthers-carburants. De plus, ce sont des biocatalyseurs très utiles qui peuvent également servir de modèles pour caractériser une réaction chimique difficile : l'activation régio-et stéréospécifique de la liaison C-H. Plusieurs autres classes d'enzymes catalysent l'oxydation des alcanes. Les souches de levures capables de dégrader les alcanes contiennent plusieurs alcanes hydroxylases appartenant à la superfamille des P450, alors que différentes bactéries contiennent des enzymes similaires au système alcane hydroxylase membranaire de Pseudomonas putida GPo1. Les alcanes à courte chaîne sont probablement oxydés par des alcanes hydroxylases solubles similaires aux méthanes monooxygénases. La présence dans l'environnement de ces oxygénases a été étudiée dans des échantillons de sols et aquifères uniquement pour les alcanes hydroxylases associés aux membranes. Abstract -Diversity of Alkane Hydroxylase Systems in the Environment

show abstract

“…In only one study using P. putida strain PpS81, which lacks alcohol dehydrogenase activity and carried the plasmids with a DNA region of about 29-kbp containing AH system genes, was the product of AH activity detected using the whole-cell reaction. 5,6) In contrast, only the long DNA region responsible for AH activity was characterized in this study.…”

mentioning

confidence: 99%

“…6) But again, AH activity was detected only indirectly, this time by measuring cooxidation of NADH. The extent of the AH reaction was very limited, possibly due to the insolubility of alkanes and enzymes in water.…”

mentioning

confidence: 99%

Biotransformation of Various Alkanes Using theEscherichia coliExpressing an Alkane Hydroxylase System fromGordoniasp. TF6

Fujii¹,

Narikawa²,

Takeda³

et al. 2004

Bioscience, Biotechnology, and Biochemistry

View full text Add to dashboard Cite

Substrate specificity of the alkane hydroxylase system of Pseudomonas oleovorans GPo1

Cited by 126 publications

References 14 publications

Kinetic characterization of the soluble butane monooxygenase from Thauera butanivorans, formerly ‘Pseudomonas butanovora’

Kinetic characterization of the soluble butane monooxygenase from Thauera butanivorans, formerly ‘Pseudomonas butanovora’

Diversity of Alkane Hydroxylase Systems in the Environment

Biotransformation of Various Alkanes Using theEscherichia coliExpressing an Alkane Hydroxylase System fromGordoniasp. TF6

Contact Info

Product

Resources

About