RESUMO
The wax ester (WE) and triacylglycerol (TAG) biosynthetic potential of marine microorganisms is poorly understood at the microbial community level. The goal of this work was to uncover the prevalence and diversity of bacteria with the potential to synthesize these neutral lipids in coastal sediments of two high latitude environments, and to characterize the gene clusters related to this process. Homolog sequences of the key enzyme, the wax ester synthase/acyl-CoA:diacylglycerol acyltransferase (WS/DGAT) were retrieved from 13 metagenomes, including subtidal and intertidal sediments of a Subantarctic environment (Ushuaia Bay, Argentina), and subtidal sediments of an Antarctic environment (Potter Cove, Antarctica). The abundance of WS/DGAT homolog sequences in the sediment metagenomes was 1.23 ± 0.42 times the abundance of 12 single-copy genes encoding ribosomal proteins, higher than in seawater (0.13 ± 0.31 times in 338 metagenomes). Homolog sequences were highly diverse, and were assigned to the Pseudomonadota, Actinomycetota, Bacteroidota and Acidobacteriota phyla. The genomic context of WS/DGAT homologs included sequences related to WE and TAG biosynthesis pathways, as well as to other related pathways such as fatty-acid metabolism, suggesting carbon recycling might drive the flux to neutral lipid synthesis. These results indicate the presence of abundant and taxonomically diverse bacterial populations with the potential to synthesize lipid storage compounds in marine sediments, relating this metabolic process to bacterial survival.
Assuntos
Diacilglicerol O-Aciltransferase , Ésteres , Regiões Antárticas , Ésteres/metabolismo , Bactérias/metabolismo , Triglicerídeos , Sedimentos GeológicosRESUMO
Undaria pinnatifida is a brown algae native to Asia that has settled in various regions worldwide, periodically contributing with large quantities of C and nutrients during its annual cycle. In this work, we analyzed a coastal site in Patagonia (Argentina) that has been colonized for three decades by U. pinnatifida, focusing on associated microbial communities in three different compartments. An important influence of algae was observed in seawater, especially in the bottom of the algal forest during the austral summer (January) at the moment of greater biomass release. This was evidenced by changes in DOC concentration and its quality indicators (higher Freshness and lower Humification index) and higher DIC. Although maximum values of NH4 and PO4 were observed in January, bottom water samples had lower concentrations than surface water, suggesting nutrient consumption by bacteria during algal DOM release. Concomitantly, bacterial abundance peaked, reaching 4.68 ± 1.33 × 105 cells mL -1 (January), showing also higher capability of degrading alginate, a major component of brown algae cell walls. Microbial community structure was influenced by sampling date, season, sampling zone (surface or bottom), and environmental factors (temperature, salinity, pH, dissolved oxygen, nutrients). Samples of epiphytic biofilms showed a distinct community structure compared to seawater, lower diversity, and remarkably high alginolytic capability, suggesting adaptation to degrade algal biomass. A high microdiversity of populations of the genus Leucothrix (Gammaproteobacteria, Thiotrichales) that accounted for a large fraction of epiphytic communities was observed, and changed over time. Epiphytic assemblages shared more taxa with bottom than with surface seawater assemblages, indicating a certain level of exchange between communities in the forest surroundings. This work provides insight into the impact of U. pinnatifida decay on seawater quality, and the role of microbial communities on adapting to massive biomass inputs through rapid DOM turnover.
Assuntos
Kelp , Undaria , Bactérias/metabolismo , Chile , Água do Mar/química , Água/metabolismoRESUMO
The aim of this work was to characterize the microbial nitrogen cycling potential in sediments from Ushuaia Bay, a subantarctic environment that has suffered a recent explosive demographic growth. Subtidal sediment samples were retrieved in triplicate from two urban points in the Bay, and analyzed through metagenomic shotgun sequencing. Sequences assigned to genes related to nitrification, nitrate reduction and denitrification were predominant in this environment with respect to metagenomes from other environments, including other marine sediments. The nosZ gene, responsible for nitrous oxide transformation into di-nitrogen, presented a high diversity. The majority of NosZ sequences were classified as Clade II (atypical) variants affiliated to different bacterial lineages such as Bacteroidetes, Chloroflexi, Firmicutes, Proteobacteria, Verrucomicrobia, as well as to Archaea. The analysis of a fosmid metagenomic library from the same site showed that the genomic context of atypical variants was variable, and was accompanied by distinct regulatory elements, suggesting the evolution of differential ecophysiological roles. This work increases our understanding of the microbial ecology of nitrogen transformations in cold coastal environments and provides evidence of an enhanced denitrification potential in impacted sediment microbial communities. In addition, it highlights the role of yet overlooked populations in the mitigation of environmentally harmful forms of nitrogen.
Assuntos
Archaea/genética , Bactérias/genética , Desnitrificação/genética , Variação Genética , Sedimentos Geológicos/microbiologia , Metagenômica , Archaea/isolamento & purificação , Proteínas Arqueais/classificação , Proteínas Arqueais/genética , Proteínas Arqueais/metabolismo , Argentina , Bactérias/isolamento & purificação , Proteínas de Bactérias/classificação , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Baías , Análise por Conglomerados , DNA/química , DNA/isolamento & purificação , DNA/metabolismo , Óxido Nitroso/metabolismo , Filogenia , Análise de Sequência de DNARESUMO
In this work, we analyzed the community structure and metabolic potential of sediment microbial communities in high-latitude coastal environments subjected to low to moderate levels of chronic pollution. Subtidal sediments from four low-energy inlets located in polar and subpolar regions from both Hemispheres were analyzed using large-scale 16S rRNA gene and metagenomic sequencing. Communities showed high diversity (Shannon's index 6.8 to 10.2), with distinct phylogenetic structures (<40% shared taxa at the Phylum level among regions) but similar metabolic potential in terms of sequences assigned to KOs. Environmental factors (mainly salinity, temperature, and in less extent organic pollution) were drivers of both phylogenetic and functional traits. Bacterial taxa correlating with hydrocarbon pollution included families of anaerobic or facultative anaerobic lifestyle, such as Desulfuromonadaceae, Geobacteraceae, and Rhodocyclaceae. In accordance, biomarker genes for anaerobic hydrocarbon degradation (bamA, ebdA, bcrA, and bssA) were prevalent, only outnumbered by alkB, and their sequences were taxonomically binned to the same bacterial groups. BssA-assigned metagenomic sequences showed an extremely wide diversity distributed all along the phylogeny known for this gene, including bssA sensu stricto, nmsA, assA, and other clusters from poorly or not yet described variants. This work increases our understanding of microbial community patterns in cold coastal sediments, and highlights the relevance of anaerobic hydrocarbon degradation processes in subtidal environments.
Assuntos
Bactérias/genética , Bactérias/metabolismo , Sedimentos Geológicos/microbiologia , Hidrocarbonetos/metabolismo , Anaerobiose , Bactérias/classificação , Bactérias/isolamento & purificação , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Biodegradação Ambiental , Clima Frio , DNA Bacteriano/genética , Metagenômica , Microbiota , Filogenia , RNA Ribossômico 16S/genéticaRESUMO
The goal of this work was to identify sequences encoding monooxygenase biocatalysts with novel features by in silico mining an assembled metagenomic dataset of polar and subpolar marine sediments. The targeted enzyme sequences were Baeyer-Villiger and bacterial cytochrome P450 monooxygenases (CYP153). These enzymes have wide-ranging applications, from the synthesis of steroids, antibiotics, mycotoxins and pheromones to the synthesis of monomers for polymerization and anticancer precursors, due to their extraordinary enantio-, regio-, and chemo- selectivity that are valuable features for organic synthesis. Phylogenetic analyses were used to select the most divergent sequences affiliated to these enzyme families among the 264 putative monooxygenases recovered from the ~14 million protein-coding sequences in the assembled metagenome dataset. Three-dimensional structure modeling and docking analysis suggested features useful in biotechnological applications in five metagenomic sequences, such as wide substrate range, novel substrate specificity or regioselectivity. Further analysis revealed structural features associated with psychrophilic enzymes, such as broader substrate accessibility, larger catalytic pockets or low domain interactions, suggesting that they could be applied in biooxidations at room or low temperatures, saving costs inherent to energy consumption. This work allowed the identification of putative enzyme candidates with promising features from metagenomes, providing a suitable starting point for further developments.
Assuntos
Metagenoma/genética , Oxigenases de Função Mista/genética , Sequência de Aminoácidos , Bactérias/genética , Biocatálise , Biotecnologia/métodos , Temperatura Baixa , Sistema Enzimático do Citocromo P-450/genética , Cinética , Oxirredução , Filogenia , Alinhamento de Sequência , Especificidade por Substrato/genéticaRESUMO
Alginates are abundant polysaccharides in brown algae that constitute an important energy source for marine heterotrophic bacteria. Despite the key role of alginate degradation processes in the marine carbon cycle, little information is available on the bacterial populations involved in these processes. The aim of this work was to gain a better understanding of alginate utilization capabilities in cold coastal environments. Sediment metagenomes from four high-latitude regions of both Hemispheres were interrogated for alginate lyase gene homologue sequences and their genomic context. Sediments contained highly abundant and diverse bacterial assemblages with alginolytic potential, including members of Bacteroidetes and Proteobacteria, as well as several poorly characterized taxa. The microbial communities in Arctic and Antarctic sediments exhibited the most similar alginolytic profiles, whereas brackish sediments showed distinct structures with a higher proportion of novel genes. Examination of the gene neighbourhood of the alginate lyase homologues revealed distinct patterns depending on the potential lineage of the scaffolds, with evidence of evolutionary relationships among alginolytic gene clusters from Bacteroidetes and Proteobacteria. This information is relevant for understanding carbon fluxes in cold coastal environments and provides valuable information for the development of biotechnological applications from brown algae biomass.
Assuntos
Alginatos/metabolismo , Bactérias/genética , Bactérias/metabolismo , Sedimentos Geológicos/microbiologia , Metagenômica , Regiões Antárticas , Regiões Árticas , Ecossistema , Ácido Glucurônico/metabolismo , Ácidos Hexurônicos/metabolismo , Metagenoma , Polissacarídeo-Liases , Polissacarídeos/metabolismoRESUMO
We aimed to gain insight into the alkane degradation potential of microbial communities from chronically polluted sediments of a subantarctic coastal environment using a combination of metagenomic approaches. A total of 6178 sequences annotated as alkane-1-monooxygenases (EC 1.14.15.3) were retrieved from a shotgun metagenomic dataset that included two sites analyzed in triplicate. The majority of the sequences binned with AlkB described in Bacteroidetes (32 ± 13 %) or Proteobacteria (29 ± 7 %), although a large proportion remained unclassified at the phylum level. Operational taxonomic unit (OTU)-based analyses showed small differences in AlkB distribution among samples that could be correlated with alkane concentrations, as well as with site-specific variations in pH and salinity. A number of low-abundance OTUs, mostly affiliated with Actinobacterial sequences, were found to be only present in the most contaminated samples. On the other hand, the molecular screening of a large-insert metagenomic library of intertidal sediments from one of the sampling sites identified two genomic fragments containing novel alkB gene sequences, as well as various contiguous genes related to lipid metabolism. Both genomic fragments were affiliated with the phylum Planctomycetes, and one could be further assigned to the genus Rhodopirellula due to the presence of a partial sequence of the 23S ribosomal RNA (rRNA) gene. This work highlights the diversity of bacterial groups contributing to the alkane degradation potential and reveals patterns of functional diversity in relation with environmental stressors in a chronically polluted, high-latitude coastal environment. In addition, alkane biodegradation genes are described for the first time in members of Planctomycetes.
Assuntos
Alcanos/metabolismo , Bactérias/isolamento & purificação , Bactérias/metabolismo , Sedimentos Geológicos/microbiologia , Poluentes Químicos da Água/metabolismo , Bactérias/classificação , Bactérias/genética , Biodegradação Ambiental , Biodiversidade , Sedimentos Geológicos/análise , Concentração de Íons de Hidrogênio , Dados de Sequência Molecular , Filogenia , Água do Mar/análise , Água do Mar/microbiologia , Poluentes Químicos da Água/análiseRESUMO
The marine environments of Argentina have a remarkable extension, as well as high biological productivity and biodiversity of both macro- and microorganisms. Despite having a great potential for biotechnological applications, the microorganisms inhabiting these ecosystems remain mostly unexplored and unexploited. In this review, we study the research topics and the interactions among Argentinean laboratories, by analyzing current articles published on biotechnology-related marine microbiology by researchers of this country. In addition, we identify the challenges and opportunities for Argentina to take advantage of the genetic potential of its marine microorganisms. Finally, we suggest possible actions that could improve the development of this research field, as well as the utilization of this knowledge to solve societal needs.
Assuntos
Biotecnologia , Biologia Marinha , Microbiologia da Água , Regiões Antárticas , Argentina , Biodegradação Ambiental , Biodiversidade , Descoberta de Drogas , Previsões , Cooperação Internacional , Consórcios Microbianos , Oceanos e Mares , Pesquisa/organização & administraçãoRESUMO
The marine environments of Argentina have a remarkable extension, as well as high biological productivity and biodiversity of both macro- and microorganisms. Despite having a great potential for biotechnological applications, the microorganisms inhabiting these ecosystems remain mostly unexplored and unexploited. In this review, we study the research topics and the interactions among Argentinean laboratories, by analyzing current articles published on biotechnology-related marine microbiology by researchers of this country. In addition, we identify the challenges and opportunities for Argentina to take advantage of the genetic potential of its marine microorganisms. Finally, we suggest possible actions that could improve the development of this research field, as well as the utilization of this knowledge to solve societal needs.
El medio ambiente marino de la Argentina tiene una notable extensión, como así también una alta productividad biológica y biodiversidad de macro y microorganismos. A pesar de presentar un gran potencial para aplicaciones biotecnológicas, los microorganismos que habitan estos ecosistemas permanecen mayormente inexplorados y sus propiedades aún no explotadas. En este trabajo de revisión, estudiamos los temas de investigación y las interacciones entre grupos de investigación argentinos, por medio del análisis de los artículos publicados hasta el momento en temáticas relacionadas con la aplicación biotecnológica de microorganismos marinos. Además, identificamos los desafíos y las oportunidades para que la Argentina tome ventaja del potencial genético de sus microorganismos marinos. Por último, sugerimos posibles acciones que podrían mejorar el desarrollo de este campo de estudio, como así también la utilización de este conocimiento para resolver las necesidades de la sociedad.
Assuntos
Biotecnologia , Biologia Marinha , Microbiologia da Água , Regiões Antárticas , Argentina , Biodegradação Ambiental , Biodiversidade , Descoberta de Drogas , Previsões , Cooperação Internacional , Consórcios Microbianos , Oceanos e Mares , Pesquisa/organização & administraçãoRESUMO
The marine environments of Argentina have a remarkable extension, as well as high biological productivity and biodiversity of both macro- and microorganisms. Despite having a great potential for biotechnological applications, the microorganisms inhabiting these ecosystems remain mostly unexplored and unexploited. In this review, we study the research topics and the interactions among Argentinean laboratories, by analyzing current articles published on biotechnology-related marine microbiology by researchers of this country. In addition, we identify the challenges and opportunities for Argentina to take advantage of the genetic potential of its marine microorganisms. Finally, we suggest possible actions that could improve the development of this research field, as well as the utilization of this knowledge to solve societal needs.(AU)
El medio ambiente marino de la Argentina tiene una notable extensión, como así también una alta productividad biológica y biodiversidad de macro y microorganismos. A pesar de presentar un gran potencial para aplicaciones biotecnológicas, los microorganismos que habitan estos ecosistemas permanecen mayormente inexplorados y sus propiedades aún no explotadas. En este trabajo de revisión, estudiamos los temas de investigación y las interacciones entre grupos de investigación argentinos, por medio del análisis de los artículos publicados hasta el momento en temáticas relacionadas con la aplicación biotecnológica de microorganismos marinos. Además, identificamos los desafíos y las oportunidades para que la Argentina tome ventaja del potencial genético de sus microorganismos marinos. Por último, sugerimos posibles acciones que podrían mejorar el desarrollo de este campo de estudio, como así también la utilización de este conocimiento para resolver las necesidades de la sociedad.(AU)
Assuntos
Biotecnologia , Biologia Marinha , Microbiologia da Água , Regiões Antárticas , Argentina , Biodegradação Ambiental , Biodiversidade , Descoberta de Drogas , Previsões , Cooperação Internacional , Consórcios Microbianos , Oceanos e Mares , Pesquisa/organização & administraçãoRESUMO
Environmental microorganisms constitute an almost inexhaustible reserve of genetic and functional diversity, accumulated during millions of years of adaptive evolution to various selective pressures. In particular, the extent of microbial biodiversity in marine habitats seems to grow larger as new techniques emerge to measure it. This has resulted in novel and more complex approaches for the screening of molecules and activities of biotechnological interest in these environments. In this review, we explore the different partially overlapping biotechnological fields that make use of microorganisms and we describe the different marine habitats that are particularly attractive for bioprospection. In addition, we review the methodological approaches currently used for microbial bioprospection, from the traditional cultivation techniques to state of the art metagenomic approaches, with emphasis in the marine environment.
Assuntos
Biotecnologia/métodos , Microbiologia Ambiental , Biologia Marinha/métodos , Inoculantes Agrícolas , Argentina , Biocatálise , Biodiversidade , Biotecnologia/tendências , Descoberta de Drogas , Ecossistema , Microbiologia Industrial/métodos , Microbiologia Industrial/tendências , Biologia Marinha/tendências , Metagenômica/métodos , Consórcios MicrobianosRESUMO
Although sediments are the natural hydrocarbon sink in the marine environment, the ecology of hydrocarbon-degrading bacteria in sediments is poorly understood, especially in cold regions. We studied the diversity of alkane-degrading bacterial populations and their response to oil exposure in sediments of a chronically polluted Subantarctic coastal environment, by analyzing alkane monooxygenase (alkB) gene libraries. Sequences from the sediment clone libraries were affiliated with genes described in Proteobacteria and Actinobacteria, with 67 % amino acid identity in average to sequences from isolated microorganisms. The majority of the sequences were most closely related to uncultured microorganisms from cold marine sediments or soils from high latitude regions, highlighting the role of temperature in the structuring of this bacterial guild. The distribution of alkB sequences among samples of different sites and years, and selection after experimental oil exposure allowed us to identify ecologically relevant alkB genes in Subantarctic sediments, which could be used as biomarkers for alkane biodegradation in this environment. 16 S rRNA amplicon pyrosequencing indicated the abundance of several genera for which no alkB genes have yet been described (Oleispira, Thalassospira) or that have not been previously associated with oil biodegradation (Spongiibacter-formerly Melitea-, Maribius, Robiginitomaculum, Bizionia and Gillisia). These genera constitute candidates for future work involving identification of hydrocarbon biodegradation pathway genes.
Assuntos
Alcanos/metabolismo , Citocromo P-450 CYP4A/genética , Poluentes Ambientais/metabolismo , Gammaproteobacteria/genética , Sedimentos Geológicos/microbiologia , Petróleo/metabolismo , Regiões Antárticas , Biodegradação Ambiental , DNA Bacteriano/genética , DNA Bacteriano/metabolismo , Gammaproteobacteria/isolamento & purificação , Gammaproteobacteria/metabolismo , Hidrocarbonetos/metabolismo , Dados de Sequência Molecular , Filogenia , Reação em Cadeia da Polimerase , RNA Ribossômico 16S/genética , Análise de Sequência de DNA/métodosRESUMO
Bioprospección de microorganismos marinos: aplicaciones biotecnológicas y métodos. Los microorganismos ambientales constituyen una reserva prácticamente inagotable de diversidad genética, acumulada durante millones de años de evolución adaptativa a varias presiones selectivas. En particular, la magnitud de la biodiversidad microbiana en hábitats marinos parece crecer al emerger nuevas técnicas para medirla. Como resultado, se han comenzado a utilizar enfoques novedosos y más complejos para la búsqueda de moléculas y actividades de interés biotecnológico en estos ambientes. En este artículo de revisión, nosotros exploramos los diferentes campos de la biotecnología que utilizan microorganismos, los cuales se superponen parcialmente, y describimos los diferentes hábitats marinos que resultan particularmente atractivos para la bioprospección. Además, revisamos los enfoques metodológicos actualmente utilizados para la bioprospección microbiana, desde las técnicas de cultivo tradicionales hasta modernos enfoques metagenómicos, con énfasis en el medio ambiente marino.
Environmental microorganisms constitute an almost inexhaustible reserve of genetic and functional diversity, accumulated during millions of years of adaptive evolution to various selective pressures. In particular, the extent of microbial biodiversity in marine habitats seems to grow larger as new techniques emerge to measure it. This has resulted in novel and more complex approaches for the screening of molecules and activities of biotechnological interest in these environments. In this review, we explore the different partially overlapping biotechnological fields that make use of microorganisms and we describe the different marine habitats that are particularly attractive for bioprospection. In addition, we review the methodological approaches currently used for microbial bioprospection, from the traditional cultivation techniques to state of the art metagenomic approaches, with emphasis in the marine environment.
Assuntos
Biotecnologia/métodos , Microbiologia Ambiental , Biologia Marinha/métodos , Inoculantes Agrícolas , Argentina , Biocatálise , Biodiversidade , Biotecnologia/tendências , Descoberta de Drogas , Ecossistema , Microbiologia Industrial/métodos , Microbiologia Industrial/tendências , Consórcios Microbianos , Biologia Marinha/tendências , Metagenômica/métodosRESUMO
Bioprospección de microorganismos marinos: aplicaciones biotecnológicas y métodos. Los microorganismos ambientales constituyen una reserva prácticamente inagotable de diversidad genética, acumulada durante millones de años de evolución adaptativa a varias presiones selectivas. En particular, la magnitud de la biodiversidad microbiana en hábitats marinos parece crecer al emerger nuevas técnicas para medirla. Como resultado, se han comenzado a utilizar enfoques novedosos y más complejos para la búsqueda de moléculas y actividades de interés biotecnológico en estos ambientes. En este artículo de revisión, nosotros exploramos los diferentes campos de la biotecnología que utilizan microorganismos, los cuales se superponen parcialmente, y describimos los diferentes hábitats marinos que resultan particularmente atractivos para la bioprospección. Además, revisamos los enfoques metodológicos actualmente utilizados para la bioprospección microbiana, desde las técnicas de cultivo tradicionales hasta modernos enfoques metagenómicos, con énfasis en el medio ambiente marino.(AU)
Environmental microorganisms constitute an almost inexhaustible reserve of genetic and functional diversity, accumulated during millions of years of adaptive evolution to various selective pressures. In particular, the extent of microbial biodiversity in marine habitats seems to grow larger as new techniques emerge to measure it. This has resulted in novel and more complex approaches for the screening of molecules and activities of biotechnological interest in these environments. In this review, we explore the different partially overlapping biotechnological fields that make use of microorganisms and we describe the different marine habitats that are particularly attractive for bioprospection. In addition, we review the methodological approaches currently used for microbial bioprospection, from the traditional cultivation techniques to state of the art metagenomic approaches, with emphasis in the marine environment.(AU)
Assuntos
Biotecnologia/métodos , Microbiologia Ambiental , Biologia Marinha/métodos , Inoculantes Agrícolas , Argentina , Biocatálise , Biodiversidade , Biotecnologia/tendências , Descoberta de Drogas , Ecossistema , Microbiologia Industrial/métodos , Microbiologia Industrial/tendências , Biologia Marinha/tendências , Metagenômica/métodos , Consórcios MicrobianosRESUMO
Bioprospección de microorganismos marinos: aplicaciones biotecnológicas y métodos. Los microorganismos ambientales constituyen una reserva prácticamente inagotable de diversidad genética, acumulada durante millones de años de evolución adaptativa a varias presiones selectivas. En particular, la magnitud de la biodiversidad microbiana en hábitats marinos parece crecer al emerger nuevas técnicas para medirla. Como resultado, se han comenzado a utilizar enfoques novedosos y más complejos para la búsqueda de moléculas y actividades de interés biotecnológico en estos ambientes. En este artículo de revisión, nosotros exploramos los diferentes campos de la biotecnología que utilizan microorganismos, los cuales se superponen parcialmente, y describimos los diferentes hábitats marinos que resultan particularmente atractivos para la bioprospección. Además, revisamos los enfoques metodológicos actualmente utilizados para la bioprospección microbiana, desde las técnicas de cultivo tradicionales hasta modernos enfoques metagenómicos, con énfasis en el medio ambiente marino.(AU)
Environmental microorganisms constitute an almost inexhaustible reserve of genetic and functional diversity, accumulated during millions of years of adaptive evolution to various selective pressures. In particular, the extent of microbial biodiversity in marine habitats seems to grow larger as new techniques emerge to measure it. This has resulted in novel and more complex approaches for the screening of molecules and activities of biotechnological interest in these environments. In this review, we explore the different partially overlapping biotechnological fields that make use of microorganisms and we describe the different marine habitats that are particularly attractive for bioprospection. In addition, we review the methodological approaches currently used for microbial bioprospection, from the traditional cultivation techniques to state of the art metagenomic approaches, with emphasis in the marine environment.(AU)
Assuntos
Biotecnologia/métodos , Microbiologia Ambiental , Biologia Marinha/métodos , Argentina , Biocatálise , Biodiversidade , Biotecnologia/tendências , Descoberta de Drogas , Ecossistema , Microbiologia Industrial/métodos , Microbiologia Industrial/tendências , Biologia Marinha/tendências , Metagenômica/métodos , Consórcios MicrobianosRESUMO
Novel polycyclic aromatic hydrocarbon dioxygenase gene variants were present in abundances similar to or higher than those of phnA1 from Cycloclasticus spp. at a chronically polluted subantarctic coastal marine environment in Patagonia. These novel gene variants were detected over a 6-year time span and were also present in sediments from temperate Patagonian sites.
Assuntos
Dioxigenases/genética , Dioxigenases/metabolismo , Sedimentos Geológicos/microbiologia , Metagenoma , Hidrocarbonetos Policíclicos Aromáticos/metabolismo , Argentina , Chile , FilogeografiaRESUMO
Biosurfactant-producing bacteria belonging to the genera Alcanivorax, Cobetia and Halomonas were isolated from marine sediments with a history of hydrocarbon exposure (Aristizábal and Gravina Peninsulas, Argentina). Two Alcanivorax isolates were found to form naturally occurring consortia with strains closely related to Pseudomonas putida and Microbacterium esteraromaticum. Alkane hydroxylase gene analysis in these two Alcanivorax strains resulted in the identification of two novel alkB genes, showing 86% and 60% deduced amino acid sequence identity with those of Alcanivorax sp. A-11-3 and Alcanivorax dieselolei P40, respectively. In addition, a gene homologous to alkB2 from Alcanivorax borkumensis was present in one of the strains. The consortium formed by this strain, Alcanivorax sp. PA2 (98.9% 16S rRNA gene sequence identity with A. borkumensis SK2(T)) and P. putida PA1 was characterized in detail. These strains form cell aggregates when growing as mixed culture, though only PA2 was responsible for biosurfactant activity. During exponential growth phase of PA2, cells showed high hydrophobicity and adherence to hydrocarbon droplets. Biosurfactant production was only detectable at late growth and stationary phases, suggesting that it is not involved in initiating oil degradation and that direct interfacial adhesion is the main hydrocarbon accession mode of PA2. This strain could be useful for biotechnological applications due to its biosurfactant production, catabolic and aggregation properties.
Assuntos
Alcanivoraceae/genética , Alcanivoraceae/isolamento & purificação , Citocromo P-450 CYP4A/metabolismo , Hidrocarbonetos/metabolismo , Tensoativos/metabolismo , Alcanivoraceae/metabolismo , Argentina , Citocromo P-450 CYP4A/genética , Genes Bacterianos , Genes de RNAr , Sedimentos Geológicos/microbiologia , Filogenia , RNA Bacteriano/genética , RNA Ribossômico 16S/genética , Água do Mar/microbiologia , Análise de Sequência de DNARESUMO
BACKGROUND: Polycyclic aromatic hydrocarbons (PAHs), widespread pollutants in the marine environment, can produce adverse effects in marine organisms and can be transferred to humans through seafood. Our knowledge of PAH-degrading bacterial populations in the marine environment is still very limited, and mainly originates from studies of cultured bacteria. In this work, genes coding catabolic enzymes from PAH-biodegradation pathways were characterized in coastal sediments of Patagonia with different levels of PAH contamination. RESULTS: Genes encoding for the catalytic alpha subunit of aromatic ring-hydroxylating dioxygenases (ARHDs) were amplified from intertidal sediment samples using two different primer sets. Products were cloned and screened by restriction fragment length polymorphism analysis. Clones representing each restriction pattern were selected in each library for sequencing. A total of 500 clones were screened in 9 gene libraries, and 193 clones were sequenced. Libraries contained one to five different ARHD gene types, and this number was correlated with the number of PAHs found in the samples above the quantification limit (r = 0.834, p < 0.05). Overall, eight different ARHD gene types were detected in the sediments. In five of them, their deduced amino acid sequences formed deeply rooted branches with previously described ARHD peptide sequences, exhibiting less than 70% identity to them. They contain consensus sequences of the Rieske type [2Fe-2S] cluster binding site, suggesting that these gene fragments encode for ARHDs. On the other hand, three gene types were closely related to previously described ARHDs: archetypical nahAc-like genes, phnAc-like genes as identified in Alcaligenes faecalis AFK2, and phnA1-like genes from marine PAH-degraders from the genus Cycloclasticus. CONCLUSION: These results show the presence of hitherto unidentified ARHD genes in this sub-Antarctic marine environment exposed to anthropogenic contamination. This information can be used to study the geographical distribution and ecological significance of bacterial populations carrying these genes, and to design molecular assays to monitor the progress and effectiveness of remediation technologies.