Abstract:Rhodococcus sp. 1BN was isolated from a contaminated site and showed various biodegradative capabilities. Besides naphthalene, strain 1BN degraded medium- (C6) and long-chain alkanes (C16-C28), benzene and toluene, alone or when the hydrocarbons were mixed in equal proportions. The nucleotide sequence of an alk polymerase chain reaction (PCR) fragment revealed a 59% nucleotide homology to the Pseudomonas oleovorans alkB gene. The nar fragments were highly homologous to genes coding for large and small subunits… Show more
“…Recent advances in characterizing alkane metabolism in a number of organisms have allowed the production of a variety of primers to detect, for example, the alkB gene from P. putida GPo1 (573). As more strains are tested and more probes are produced, it is becoming clear that, while different alkane hydroxylases can be found in phylogenetically distant microorganisms (19), many probes will only provide information on the presence of a similar gene in closely related strains. Thus, the usefulness of such gene probes will grow as the diversity of genes responsible for hy-VOL.…”
Recent advances in molecular biology have extended our understanding of the metabolic processes related to microbial transformation of petroleum hydrocarbons. The physiological responses of microorganisms to the presence of hydrocarbons, including cell surface alterations and adaptive mechanisms for uptake and efflux of these substrates, have been characterized. New molecular techniques have enhanced our ability to investigate the dynamics of microbial communities in petroleum-impacted ecosystems. By establishing conditions which maximize rates and extents of microbial growth, hydrocarbon access, and transformation, highly accelerated and bioreactor-based petroleum waste degradation processes have been implemented. Biofilters capable of removing and biodegrading volatile petroleum contaminants in air streams with short substrate-microbe contact times (<60 s) are being used effectively. Microbes are being injected into partially spent petroleum reservoirs to enhance oil recovery. However, these microbial processes have not exhibited consistent and effective performance, primarily because of our inability to control conditions in the subsurface environment. Microbes may be exploited to break stable oilfield emulsions to produce pipeline quality oil. There is interest in replacing physical oil desulfurization processes with biodesulfurization methods through promotion of selective sulfur removal without degradation of associated carbon moieties. However, since microbes require an environment containing some water, a two-phase oil-water system must be established to optimize contact between the microbes and the hydrocarbon, and such an emulsion is not easily created with viscous crude oil. This challenge may be circumvented by application of the technology to more refined gasoline and diesel substrates, where aqueous-hydrocarbon emulsions are more easily generated. Molecular approaches are being used to broaden the substrate specificity and increase the rates and extents of desulfurization. Bacterial processes are being commercialized for removal of H2S and sulfoxides from petrochemical waste streams. Microbes also have potential for use in removal of nitrogen from crude oil leading to reduced nitric oxide emissions provided that technical problems similar to those experienced in biodesulfurization can be solved. Enzymes are being exploited to produce added-value products from petroleum substrates, and bacterial biosensors are being used to analyze petroleum-contaminated environments
“…Recent advances in characterizing alkane metabolism in a number of organisms have allowed the production of a variety of primers to detect, for example, the alkB gene from P. putida GPo1 (573). As more strains are tested and more probes are produced, it is becoming clear that, while different alkane hydroxylases can be found in phylogenetically distant microorganisms (19), many probes will only provide information on the presence of a similar gene in closely related strains. Thus, the usefulness of such gene probes will grow as the diversity of genes responsible for hy-VOL.…”
Recent advances in molecular biology have extended our understanding of the metabolic processes related to microbial transformation of petroleum hydrocarbons. The physiological responses of microorganisms to the presence of hydrocarbons, including cell surface alterations and adaptive mechanisms for uptake and efflux of these substrates, have been characterized. New molecular techniques have enhanced our ability to investigate the dynamics of microbial communities in petroleum-impacted ecosystems. By establishing conditions which maximize rates and extents of microbial growth, hydrocarbon access, and transformation, highly accelerated and bioreactor-based petroleum waste degradation processes have been implemented. Biofilters capable of removing and biodegrading volatile petroleum contaminants in air streams with short substrate-microbe contact times (<60 s) are being used effectively. Microbes are being injected into partially spent petroleum reservoirs to enhance oil recovery. However, these microbial processes have not exhibited consistent and effective performance, primarily because of our inability to control conditions in the subsurface environment. Microbes may be exploited to break stable oilfield emulsions to produce pipeline quality oil. There is interest in replacing physical oil desulfurization processes with biodesulfurization methods through promotion of selective sulfur removal without degradation of associated carbon moieties. However, since microbes require an environment containing some water, a two-phase oil-water system must be established to optimize contact between the microbes and the hydrocarbon, and such an emulsion is not easily created with viscous crude oil. This challenge may be circumvented by application of the technology to more refined gasoline and diesel substrates, where aqueous-hydrocarbon emulsions are more easily generated. Molecular approaches are being used to broaden the substrate specificity and increase the rates and extents of desulfurization. Bacterial processes are being commercialized for removal of H2S and sulfoxides from petrochemical waste streams. Microbes also have potential for use in removal of nitrogen from crude oil leading to reduced nitric oxide emissions provided that technical problems similar to those experienced in biodesulfurization can be solved. Enzymes are being exploited to produce added-value products from petroleum substrates, and bacterial biosensors are being used to analyze petroleum-contaminated environments
“…Comparison of the three alkane hydroxylase sequences showed that four histidine-containing motifs were well conserved, three of which were also present in the distantly related membranebound desaturases [51]. Based on two of the motifs, highly degenerate primers were developed that amplified internal gene fragments of alkB homologs from gram-negative as well as gram-positive strains [32,50,53,69] (Table 3). In the initial PCR study, most strains that were able to grow on alkanes yielded alkB gene fragments [50], and the same was found for a set of Rhodococcus strains isolated from soil in Bremen, Germany [86,87].…”
Section: Diversity Of Membrane-bound Alkane Hydroxylase Systemsmentioning
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
“…Sequencing of these PCR products after cloning confirmed their homology to alkB. Derived protein sequences showed highest homologies to AlkB from other bacterial gene (Smits et al, 1999;Khalameyzer et al, 1999;Andreoni et al, 2000;Kloos et al, 2006) …”
Section: Evaluation Of the Detection Methods With The Reference Isolatesmentioning
Biodegradation is the chemical breakdown of materials by a physiological environment. The term is often used in relation to ecology, waste management and environmental remediation. A specific gene from bacteria is identified and sequenced which has the capacity of oil degradation without any obvious strain specific discrimination using a combination of PCR and hybridization. The parameters of biodegradation that is culturing and PCR technique provide useful information for an assessment of the intrinsic biodegradation potential that is present at a site. PCR amplification products in the plasmid DNA of Pseudomonas aeruginosa was transformed into competent Escherichia coli cells. Thus the E. coli cells were conferred with oil degrading property and this was confirmed by growing them in Bushnell Hass medium along with petroleum oil. The E. coli cells were found to be catabolizing the oil. Results show that the capability for alkane degradation is a common trait in microbial communities. The method can be a very useful tool for the fast estimation of the biodegradation potential at polluted sites.
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