Molecular analysis of the bacterial diversity in a specialized consortium for diesel oil degradation

Paixão, Douglas Antônio Alvaredo; Dimitrov, Maurício R.; Pereira, Rodrigo Matheus; Accorsini, Fábio Raphael; Vidotti, Maria Benincasa; Lemos, Eliana Gertrudes de Macedo

doi:10.1590/s0100-06832010000300019

Cited by 16 publications

(9 citation statements)

References 43 publications

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“…As in a previous work, a soil sample was collected from a region contaminated with petroleum hydrocarbons from a factory of automotive lubricants in the city of Ribeirão Preto, São Paulo State, Brazil [ 21 ]. The soil was used to develop a microbial consortium at the Department of Biology at the University of São Paulo State, Jaboticabal Campus [ 21 ], which had the bacterial diversity analyzed [ 22 ]. From this microbial consortium, a metagenomic library of fosmid vectors was built in the Biochemistry Laboratory of Microorganisms and Plants (LBMP) in the Department of Technology at the same university.…”

Section: Methodsmentioning

confidence: 99%

Est16, a New Esterase Isolated from a Metagenomic Library of a Microbial Consortium Specializing in Diesel Oil Degradation

et al. 2015

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Lipolytic enzymes have attracted attention from a global market because they show enormous biotechnological potential for applications such as detergent production, leather processing, cosmetics production, and use in perfumes and biodiesel. Due to the intense demand for biocatalysts, a metagenomic approach provides methods of identifying new enzymes. In this study, an esterase designated as Est16 was selected from 4224 clones of a fosmid metagenomic library, revealing an 87% amino acid identity with an esterase/lipase (accession number ADM63076.1) from an uncultured bacterium. Phylogenetic studies showed that the enzyme belongs to family V of bacterial lipolytic enzymes and has sequence and structural similarities with an aryl-esterase from Pseudomonas fluorescens and a patented Anti-Kazlauskas lipase (patent number US20050153404). The protein was expressed and purified as a highly soluble, thermally stable enzyme that showed a preference for basic pH. Est16 exhibited activity toward a wide range of substrates and the highest catalytic efficiency against p-nitrophenyl butyrate and p-nitrophenyl valerate. Est16 also showed tolerance to the presence of organic solvents, detergents and metals. Based on molecular modeling, we showed that the large alpha-beta domain is conserved in the patented enzymes but not the substrate pocket. Here, it was demonstrated that a metagenomic approach is suitable for discovering the lipolytic enzyme diversity and that Est16 has the biotechnological potential for use in industrial processes.

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Section: Methodsmentioning

confidence: 99%

Est16, a New Esterase Isolated from a Metagenomic Library of a Microbial Consortium Specializing in Diesel Oil Degradation

et al. 2015

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“…This strain was isolated from an activated sludge for its ability to degrade linear alkylbenzene sulfonate surfactants (Schleheck et al 2004(Schleheck et al , 2007. The genus Parvibaculum includes hydrocarbon-degrading species, which were detected in several hydrocarbon-contaminated environments (Alonso-Gutiérrez et al 2009;Paixão et al 2010;Wang et al 2010). Consequently, the enrichment of genus Parvibaculum suggests that the hydrocarbon-degrading and/or biosurfactants-producing bacteria may exist in the microcosms treated with crude oil.…”

Section: Phylogenetic Analysis Of Major Dgge Bands After 30 Days Of Imentioning

confidence: 99%

Synergistic effect of crude oil plus dispersant on bacterial community in a louisiana salt marsh sediment

Al-Jawasim¹,

Yu²,

Park³

2015

FEMS Microbiology Letters

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A combined effect of crude oil plus dispersant (Corexit 9500A) significantly altered indigenous bacterial communities in a Louisiana salt marsh sediment after 30 days of incubation; the crude oil and/or Corexit 9500A treatments triggered shifts in bacterial communities and the shifted bacterial structure by crude oil plus Corexit 9500A was considerably different from those by either crude oil or Corexit 9500A. However, the synergistic effect of crude oil plus Corexit 9500A was not observed after 7 days of incubation; the bacterial community was slightly shifted by Corexit 9500A and the crude oil did not trigger any bacterial community shift after 7 days of incubation. DNA sequencing data indicated that Chromobacterium species was enriched in the Corexit 9500A microcosms after 7 days of incubation, while Pseudomonas, Advenella, Acidocella and Dyella spp. were enriched after 30 days of incubation. Parvibaculum was a dominant species in the crude oil microcosms after 30 days of incubation. Rhodanobacter, Dyella and Frateuria spp. were dominant in crude oil plus Corexit 9500A microcosms after 30 days of incubation. Our data show that the effect of crude oil plus Corexit 9500A on bacterial community is synergistic, and thus the dispersant effect should be considered with the spilled oil to correctly evaluate the environmental impact.

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“…In order to identify enzymes for different applications, a fosmid metagenomic library consisting of 4224 clones was previously constructed in the Laboratory of Biochemistry of Microorganisms and Plants at UNESP/Jaboticabal-Brazil. The soil was collected in a region contaminated with petroleum hydrocarbons from a former automotive lubricant factory in the city of Ribeirão Preto, São Paulo State, Brazil (Accorsini et al 2012), and was used to develop a microbial consortium that was specialized for diesel oil degradation (Paixão et al 2010) and for the subsequent construction of the metagenomic library. Recently, we have performed a screening for clones exhibiting lipolytic activity on agar containing tributyrin as the indicator substrate, which resulted in the identification of 30 positive clones.…”

Section: Introductionmentioning

confidence: 99%

Characterization of EST3: a metagenome-derived esterase with suitable properties for biotechnological applications

Maester

Pereira

Sierra

et al. 2016

Appl Microbiol Biotechnol

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Metagenomic libraries from diverse environments have been extensive sources of many lipases and esterases; nevertheless, most of these enzymes remain biochemically uncharacterized. We previously built a metagenomic fosmid library from a microbial consortium specialized for diesel oil degradation and tested it for lipolytic activity. In the present study, we identified the PL14.H10 clone that was subcloned and sequenced, which enabled the identification of the EST3 protein. This enzyme exhibited 74 % amino acid identity with the uncharacterized alpha/beta hydrolase from Parvibaculum lavamentivorans [GenBank: WP012110575.1] and was classified into lipolytic enzyme family IV. Biochemical characterization revealed that EST3 presents high activity in a wide range of temperature with highest activity from 41 to 45 °C. Also, this thermostable esterase acts from mild acidic to alkaline conditions with an optimum pH of 6.0. The enzyme exhibited activity against p-nitrophenyl esters of different chain lengths and highest catalytic efficiency against p-nitrophenyl caprylate. The activity of the protein was increased in the presence of 0.5 mM of Mn(+2), Li(+), EDTA, and 1 % of CTAB and exhibited half of the activity in the presence of 10 % methanol and ethanol. Moreover, the homology model of EST3 was built and compared to other esterases, revealing a substrate channel that should fit a wide range of substrates. Taken together, the data presented in this work reveal the unique and interesting characteristics of EST3 that might be explored for further use in biotechnological applications.

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Molecular analysis of the bacterial diversity in a specialized consortium for diesel oil degradation

Cited by 16 publications

References 43 publications

Est16, a New Esterase Isolated from a Metagenomic Library of a Microbial Consortium Specializing in Diesel Oil Degradation

Est16, a New Esterase Isolated from a Metagenomic Library of a Microbial Consortium Specializing in Diesel Oil Degradation

Synergistic effect of crude oil plus dispersant on bacterial community in a louisiana salt marsh sediment

Characterization of EST3: a metagenome-derived esterase with suitable properties for biotechnological applications

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