n-Alkane assimilation and tert-butyl alcohol (TBA) oxidation capacity in Mycobacterium austroafricanum strains

Ferreira, Nicolas Lopes; Mathis, Hugues; Labbé, Diane; Monot, Frédéric; Greer, Charles W.; Fayolle-Guichard, Françoise

doi:10.1007/s00253-007-0892-1

Cited by 38 publications

(32 citation statements)

References 38 publications

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“…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) . The soluble haem-containing CYP153A6 alkane hydroxylase from Mycobacterium sp.…”

Section: Substrate Specificitymentioning

confidence: 99%

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

et al. 2009

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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.

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Section: Substrate Specificitymentioning

confidence: 99%

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

et al. 2009

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“…Nevertheless, in the last two years, some AlkB hydroxylases have been reported to be induced by gaseous alkanes (C 2 -C 4 ). Mycobacterium austroafricanum can grow on C 2 -C 16 and the alkB gene expression, involved in tert-butyl alcohol (TBA) co-metabolism, was induced not only after growth on n-hexane and n-hexadecane but also after growth on propane ( [67]. The involvement of AlkB-type hydroxylase in gaseous alkanes metabolism has been also reported in Pseudomonas putida GPo1 [165].…”

Section: The Membrane-bound Protein Non-heme Iron Oxygenasesmentioning

confidence: 86%

“…TF6 alkane-1-monooxygenase [336] was 76% and to Mycobacterium austroafricanum alkane 1-monooxygenase [67] was 59%. The similarity to the Geobacillus sp.…”

Section: The Alkb Gene Cluster In Rhodococcus Sp Bcp1mentioning

confidence: 95%

“…All SDIMO groups so far examined include enzymes capable of co-metabolically oxidizing halogenated pollutants such as trichloroethene (TCE) and vinyl chloride (VC) [150][151][152][153]. Some members showed also the ability to co-metabolize methyl tert-butyl ether (MTBE) [67]. Potential applications in bioremediation and biocatalysis are a strong incentive to study the details of cometabolic oxidations by alkane hydroxylase systems.…”

Section: Significance Of Sdimo Hydroxylases In Bioremediationmentioning

confidence: 99%

“…It is important to emphasize how expanding knowledge about alkane hydroxylation systems blur the boundaries of the hydroxylase classification into enzymes oxidizing gaseous alkanes and enzymes oxidizing liquid alkanes [65]. For instance, AlkB-like monooxygenase typically oxidizes liquid alkanes but, lately, few members of this family of enzymes were described to be also induced by propane [66,67]. This underlines the diversity that can exist the proteomic analysis of its cytoplasmic and membrane proteins after the growth on nhexadecane showed the presence of multiple alkane hydroxylase systems belonging to different families [75] involved in the primary oxidation of a broad range of alkanes.…”

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Monooxygenases involved in the n-alkanes metabolism by Rhodococcus sp. BCP1: molecular characterization and expression of alkB gene

Cappelletti

Fedi

Honda

et al. 2010

Journal of Biotechnology

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Discussion 107 Summary 113Chapter 5 -Analysis of the alkB gene expression Summary 157Chapter 6 -Proteomic analysis of n-alkanes growth of Rhodococcus sp. BCP1 Chapter 1 -General Introduction PrefaceThe quality of life on Earth is tightly related with the overall quality of the environment [1]. During the last century it was commonly believed that the abundance of land and resources were unlimited and it was rarely acknowledged that the production, use, and disposal of hazardous substances had environmental and health effects [1][2][3]. Owing to this, human activity has deliberately or inadvertently released into waters and soil a variety of toxic compounds as refrigerants, paints, solvents, herbicides and pesticides that can cause considerable environmental pollution and human health problems as a result of their persistence, toxicity, and transformation into hazardous metabolites [4,5]. Very soon, however, it became clear how the carelessness and negligence in handling the industrial wastes and gas emissions had caused dramatic effect on the nature and on human health [1].Nowadays, the concern about the global warming and the climate changing induced the government regulatory agencies to establish procedures for assessing the environmental impact and health hazards of chemicals and for controlling/limiting their utilization.International efforts have been applied to remedy many contaminated sites all over the world, either as a response to the risk of adverse health or environmental effects caused by contamination or to enable the site to be redeveloped for use [5]. In response to public and government concern and because of the intriguing research problems presented, environmental scientists, biologists and chemists have been giving increased attention to identifying and determining the behavior and fate of organic compounds in natural ecosystems [5].Bioremediation procedures offer an economical and effective possibility to degrade or render innocuous toxic pollutants using natural biological activity [1]. By definition, bioremediation implies the use of living organisms, primarily microorganisms, to degrade the Since contaminant compounds are transformed by living organisms through reactions that take place as a part of their metabolic processes, environmental biotechnology focuses on the development of techniques to optimize environmental parameters to allow these metabolic pathways to proceed faster and, therefore, to improve the bio-degradation [1]. As a result, the removal of the contaminated soil can be necessary to let the biodegradation proceed in controlled sites (such as bioreactors). These ex situ approaches provide optimal conditions out one single step of the entire bio-degradation process. Hydrocarbons in the environmentHydrocarbons are energy-rich organic compounds consisting of carbon and hydrogen atoms and they can be saturated (only single C-H bounds) or unsaturated (one ore more double or triple C-H bounds). Alkanes are saturated hydrocarbons with the general formula C n H 2n+2 ; they can b...

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Biochemical characterization of two recombinant ferredoxin reductases from Alcanivorax borkumensis SK2

Teimoori

Ahmadian

Madadkar-Sobhani

2012

Biotech and App Biochem

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Alcanivorax borkumensis strain SK2 is a cosmopolitan oil-degrading oligotrophic marine γ-proteobacterium that exclusively uses petroleum hydrocarbons as sources of carbon and energy. Its ubiquity and unusual physiology suggest its global importance in the removal of hydrocarbons from polluted marine systems. The genome of A. borkumensis SK2 was recently sequenced. Two ferredoxin-nicotinamide adenine dinucleotide phosphate (NADPH) reductase genes (ABO_0145 and ABO_0203) have been annotated for this bacterium. In the present study, the expression, purification, and kinetic properties of these two genes were explored by constructing the prokaryotic expression vectors (pET21a) for the first time. Isopropyl β-D-thiogalactoside (0.5 mM) was used for induction of exponentially growing cells (30 °C, overnight). Most of the proteins were expressed in inclusion body. Partial purification of recombinant enzymes was performed by ion-exchange chromatography on a DEAE-sepharose column using only one linear gradient of sodium chloride ranging between 0 and 500 mM. The recombinant enzymes displayed reductase activity, which was optimal at pH 6.0 and 45 °C. Ferredoxin-NADPH reductases exhibited several outstanding properties that made them excellent model proteins to address broad biological questions. This study serves as the basis for further investigations of the biotechnological potential of these enzymes.

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n-Alkane assimilation and tert-butyl alcohol (TBA) oxidation capacity in Mycobacterium austroafricanum strains

Cited by 38 publications

References 38 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’

Monooxygenases involved in the n-alkanes metabolism by Rhodococcus sp. BCP1: molecular characterization and expression of alkB gene

Biochemical characterization of two recombinant ferredoxin reductases from Alcanivorax borkumensis SK2

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