RESUMO

Extracellular proteases from halophilic archaea displays increased enzymatic activities in hypersaline environment. In this study, an extracellular protease-coding gene, hly34, from the haloarchaeal strain Halococcus salifodinae PRR34, was obtained through homologous search. The protease activity produced by this strain at 20% NaCl, 42 °C, and pH 7.0 was 32.5 ± 0.5 (U·mL-1). The codon-optimized hly34 which is specific for Escherichia coli can be expressed in E. coli instead of native hly34. It exhibits proteolytic activity under a wide range of low- or high-salt concentrations, slightly acidic or alkaline conditions, and slightly higher temperatures. The Hly34 presented the highest proteolytic activity at 50 °C, pH 9.0, and 0-1 M NaCl. It was found that the Hly34 showed a higher enzyme activity under low-salt conditions. Hly34 has good stability at different NaCl concentrations (1-4 M) and pH (6.0-10.0), as well as good tolerance to some metal ions. However, at 60 °C, the stability is reduced. It has a good tolerance to some metal ions. The proteolytic activity was completely inhibited by phenylmethanesulfonyl fluoride, suggesting that the Hly34 is a serine protease. This study further deepens our understanding of haloarchaeal extracellular protease, most of which found in halophilic archaea are classified as serine proteases. These proteases exhibit a certain level of alkaline resistance and moderate heat resistance, and they may emerge with higher activity under low-salt conditions than high-salt conditions. The protease Hly34 is capable of degrading a number of proteins, including substrate proteins, such as azocasein, whey protein and casein. It has promising applications in industrial production.

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RESUMO

Two pink-pigmented halophilic archaea, designated strains ZJ1T and J81, were isolated from rock salt of Yunnan Salt Mine, China, and commercial salt imported from Bolivia, respectively. Cells were non-motile, coccoid, approximately 0.8-1.6 µm in diameter, stained Gram-negative and often occurred in pairs. Colonies were wet, opaque and smooth-edged. Strain ZJ1T grew optimally with 20 % (w/v) NaCl, at pH 7.5 and at 38-40 °C, which was the same as for strain J81. 16S rRNA gene sequence similarity between strains ZJ1T and J81 was 99.7 %. Sequence similarity searches based on the 16S rRNA gene and cell morphology suggested that strains ZJ1T and J81 belong to the genus Halococcus in the family Halococcaceae. The major polar lipids of the type strain, ZJ1T, were phosphatidylglycerol, phosphatidylglycerol phosphate methyl ester and sulfated diglycosyl-diether-1. The profile of polar lipids, cell shape, motility and lack of lysis of cells in distilled water show that strains ZJ1T and J81 were similar to other members of the genus Halococcus. Strain ZJ1T shared the highest 16S rRNA gene and rpoB' gene sequence similarities of 99.0 and 95.3 % with Halococcus hamelinensis 100A6T, respectively, followed by less than 94.6 % with sequences of other species in the genus Halococcus. DNA-DNA relatedness between strains ZJ1T and J81 was 90.1±0.7 %, while 27±0.5 % was found between strain ZJ1T and H. hamelinensis JCM 12892T (=100A6T), and 29.0±0.5 % between strains J81 and H. hamelinensis JCM 12892T. The DNA G+C content of strain ZJ1T was 66.5 mol% (Tm). The stable phylogenetic position, differential physiological and biochemical properties and extensive sequence divergence suggest that strains ZJ1T and J81 represent a novel species, for which the name Halococcus salsus sp. nov. is proposed. The type strain is ZJ1T (=CGMCC 1.16025T=NBRC 112867T).

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RESUMO

Methanogenesis and sulfate reduction are important microbial processes in hypersaline environments. However, key aspects determining substrate competition between these microbial processes have not been well documented. We evaluated competitive and non-competitive substrates for stimulation of both processes through microcosm experiments of hypersaline microbial mat samples from Guerrero Negro, Baja California Sur, Mexico, and we assessed the effect of these substrates on the microbial community composition. Methylotrophic methanogenesis evidenced by sequences belonging to methanogens of the family Methanosarcinaceae was found as the dominant methanogenic pathway in the studied hypersaline microbial mat. Nevertheless, our results showed that incubations supplemented with acetate and lactate, performed in absence of sulfate, also produced methane after 40 days of incubation, apparently driven by hydrogenotrophic methanogens affiliated to the family Methanomicrobiaceae. Sulfate reduction was mainly stimulated by addition of acetate and lactate; however, after 40 days of incubation, an increase of the H2S concentrations in microcosms amended with trimethylamine and methanol was also observed, suggesting that these substrates are putatively used for sulfate reduction. Moreover, 16S rRNA gene sequencing analysis showed remarkable differences in the microbial community composition among experimental treatments. In the analyzed sample amended with acetate, sulfate-reducing bacteria (SRB) belonging to the family Desulfobacteraceae were dominant, while members of Desulfohalobiaceae, Desulfomicrobiaceae, and Desulfovibrionaceae were found in the incubation with lactate. Additionally, we detected an unexpected high abundance of unclassified Hydrogenedentes (near 25%) in almost all the experimental treatments. This study contributes to better understand methanogenic and sulfate-reducing activities, which play an important role in the functioning of hypersaline environments.

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Fusibacter sp. 3D3 (ATCC BAA-2418) is an arsenate-reducing halotolerant strain within the Firmicutes phylum, isolated from the Salar de Ascotán, a hypersaline salt flat in Northern Chile. This high-Andean closed basin is an athalassohaline environment located at the bottom of a tectonic basin surrounded by mountain range, including some active volcanoes. This landscape can be an advantageous system to explore the effect of salinity on microorganisms that mediate biogeochemical reactions. Since 2000, microbial reduction of arsenic has been evidenced in the system, and the phylogenetic analysis of the original community plus the culture enrichments has revealed the predominance of Firmicutes phylum. Here, we describe the first whole draft genome sequence of an arsenic-reducing strain belonging to the Fusibacter genus showing the highest 16S rRNA gene sequence similarity (98%) with Fusibacter sp. strain Vns02. The draft genome consists of 57 contigs with 5,111,250 bp and an average G + C content of 37.6%. Out of 4780 total genes predicted, 4700 genes code for proteins and 80 genes for RNAs. Insights from the genome sequence and some microbiological features of the strain 3D3 are available under Bioproject accession PRJDB4973 and Biosample SAMD00055724. The release of the genome sequence of this strain could contribute to the understanding of the arsenic biogeochemistry in extreme environments.

RESUMO

Abstract This research evaluated the effect of flooding on the microphytobenthos community structure in a microbial mat from a tropical salt flat. Field samples were collected during four consecutive days: on the first three days the salt flat was dry, on the fourth day it was flooded by rain. In order to evaluate the community maintained in flood conditions, samples from this area were collected and kept in the laboratory for 10 days with sea water. The results of total abundance of microphytobenthos varied from 4.2 × 108 to 2.9 × 109 organisms L–1, total density increased one order of magnitude under the effect of water for both situations of precipitation in the salt flat and in experimental conditions, an increase due to the high abundance of Microcoleus spp. Shannon index (H’) was higher during the desiccation period. Our data suggest that changes in the abundance of organisms were due to the effect of water. The dominance of the most abundant taxa remained the same under conditions of desiccation and influence of water, and there is probably a consortium of microorganisms in the microbial mat that helps to maintain these dominances.

Resumo Esse trabalho avaliou os efeitos da inundação na estrutura da comunidade microfitobentônica de um tapete microbiano em uma planície hipersalina tropical. As amostragens foram realizadas no campo durante quatro dias consecutivos: nos três primeiros dias o local estava seco e no quarto dia foi inundando com chuva. Para avaliar a comunidade mantida em condições de inundação, foram coletadas amostras dessa região, sendo mantidas em laboratório por 10 dias com água do mar. Os resultados mostraram que a abundância total do microfitobentos variou de 4.2 × 108 a 2.9 × 109 organismos L–1, a densidade aumentou em uma ordem de grandeza com a influência da água, tanto na planície hipersalina como nas condições experimentais, um aumento que foi devido às maiores abundâncias de Microcoleus spp. O índice de Shannon (H’) foi mais elevado durante o período de dessecação. Nossos dados sugerem que as mudanças na abundância dos organismos foram devidas ao efeito da água, a dominância dos táxons mais abundantes permaneceu a mesma durante as condições de dessecação e inundação, e possivelmente existe um consórcio entre os microrganismos do tapete microbiano no qual eles ajudam a manter essa dominância.

Assuntos

RESUMO

This research evaluated the effect of flooding on the microphytobenthos community structure in a microbial mat from a tropical salt flat. Field samples were collected during four consecutive days: on the first three days the salt flat was dry, on the fourth day it was flooded by rain. In order to evaluate the community maintained in flood conditions, samples from this area were collected and kept in the laboratory for 10 days with sea water. The results of total abundance of microphytobenthos varied from 4.2 × 108 to 2.9 × 109 organisms L1, total density increased one order of magnitude under the effect of water for both situations of precipitation in the salt flat and in experimental conditions, an increase due to the high abundance of Microcoleus spp. Shannon index (H) was higher during the desiccation period. Our data suggest that changes in the abundance of organisms were due to the effect of water. The dominance of the most abundant taxa remained the same under conditions of desiccation and influence of water, and there is probably a consortium of microorganisms in the microbial mat that helps to maintain these dominances.(AU)

Esse trabalho avaliou os efeitos da inundação na estrutura da comunidade microfitobentônica de um tapete microbiano em uma planície hipersalina tropical. As amostragens foram realizadas no campo durante quatro dias consecutivos: nos três primeiros dias o local estava seco e no quarto dia foi inundando com chuva. Para avaliar a comunidade mantida em condições de inundação, foram coletadas amostras dessa região, sendo mantidas em laboratório por 10 dias com água do mar. Os resultados mostraram que a abundância total do microfitobentos variou de 4.2 × 108 a 2.9 × 109 organismos L1, a densidade aumentou em uma ordem de grandeza com a influência da água, tanto na planície hipersalina como nas condições experimentais, um aumento que foi devido às maiores abundâncias de Microcoleus spp. O índice de Shannon (H) foi mais elevado durante o período de dessecação. Nossos dados sugerem que as mudanças na abundância dos organismos foram devidas ao efeito da água, a dominância dos táxons mais abundantes permaneceu a mesma durante as condições de dessecação e inundação, e possivelmente existe um consórcio entre os microrganismos do tapete microbiano no qual eles ajudam a manter essa dominância.(AU)

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RESUMO

Isabel Lake is a moderate saline soda crater lake located in Isabel Island in the eastern tropical Pacific coast of Mexico. Lake is mainly formed by rainfall and is strongly affected by evaporation and high input of nutrients derived from excretions of a large bird community inhabiting the island. So far, only the island macrobiota has been studied. The knowledge of the prokaryotic biota inhabiting the upper layers of this meromictic lake can give clues for the maintenance of this ecosystem. We assessed the diversity and composition of prokaryotic community in sediments and water of the lake by DGGE profiling, 16S rRNA gene amplicon pyrosequencing, and cultivation techniques. The bacterial community is largely dominated by halophilic and halotolerant microorganisms. Alpha diversity estimations reveal higher value in sediments than in water (P > 0.005). The lake water is dominated by γ-Proteobacteria belonging to four main families where Halomonadaceae presents the highest abundance. Aerobic, phototrophic, and halotolerant prokaryotes such as Cyanobacteria GPIIa, Halomonas, Alcanivorax, Idiomarina, and Cyclobacterium genera are commonly found. However, in sediment samples, Formosa, Muricauda, and Salegentibacter genera corresponding to Flavobacteriaceae family accounted for 15-20 % of the diversity. Heterotrophs like those involved in sulfur cycle, Desulfotignum, Desulfuromonas, Desulfofustis, and Desulfopila, appear to play an important role in sediments. Finally, a collection of aerobic halophilic bacterial isolates was created from these samples; members of the genus Halomonas were predominantly isolated from lake water. This study contributes to state the bacterial diversity present in this particular soda saline crater lake.

Assuntos

RESUMO

Abstract This research evaluated the effect of flooding on the microphytobenthos community structure in a microbial mat from a tropical salt flat. Field samples were collected during four consecutive days: on the first three days the salt flat was dry, on the fourth day it was flooded by rain. In order to evaluate the community maintained in flood conditions, samples from this area were collected and kept in the laboratory for 10 days with sea water. The results of total abundance of microphytobenthos varied from 4.2 × 108 to 2.9 × 109 organisms L1, total density increased one order of magnitude under the effect of water for both situations of precipitation in the salt flat and in experimental conditions, an increase due to the high abundance of Microcoleus spp. Shannon index (H) was higher during the desiccation period. Our data suggest that changes in the abundance of organisms were due to the effect of water. The dominance of the most abundant taxa remained the same under conditions of desiccation and influence of water, and there is probably a consortium of microorganisms in the microbial mat that helps to maintain these dominances.

Resumo Esse trabalho avaliou os efeitos da inundação na estrutura da comunidade microfitobentônica de um tapete microbiano em uma planície hipersalina tropical. As amostragens foram realizadas no campo durante quatro dias consecutivos: nos três primeiros dias o local estava seco e no quarto dia foi inundando com chuva. Para avaliar a comunidade mantida em condições de inundação, foram coletadas amostras dessa região, sendo mantidas em laboratório por 10 dias com água do mar. Os resultados mostraram que a abundância total do microfitobentos variou de 4.2 × 108 a 2.9 × 109 organismos L1, a densidade aumentou em uma ordem de grandeza com a influência da água, tanto na planície hipersalina como nas condições experimentais, um aumento que foi devido às maiores abundâncias de Microcoleus spp. O índice de Shannon (H) foi mais elevado durante o período de dessecação. Nossos dados sugerem que as mudanças na abundância dos organismos foram devidas ao efeito da água, a dominância dos táxons mais abundantes permaneceu a mesma durante as condições de dessecação e inundação, e possivelmente existe um consórcio entre os microrganismos do tapete microbiano no qual eles ajudam a manter essa dominância.

RESUMO

Literature on hydrocarbon degradation in extreme hypersaline media presents studies that point to a negative effect of salinity increase on hydrocarbonoclastic activity, while several others report an opposite tendency. Based on information available in the literature, we present a discussion on the reasons that justify these contrary results. Despite the fact that microbial ability to metabolize hydrocarbons is found in extreme hypersaline media, indeed some factors are critical for the occurrence of hydrocarbon degradation in such environments. How these factors affect hydrocarbon degradation and their implications for the assessment of hydrocarbon biodegradation in hypersaline environments are presented in this review.

Assuntos

RESUMO

Literature on hydrocarbon degradation in extreme hypersaline media presents studies that point to a negative effect of salinity increase on hydrocarbonoclastic activity, while several others report an opposite tendency. Based on information available in the literature, we present a discussion on the reasons that justify these contrary results. Despite the fact that microbial ability to metabolize hydrocarbons is found in extreme hypersaline media, indeed some factors are critical for the occurrence of hydrocarbon degradation in such environments. How these factors affect hydrocarbon degradation and their implications for the assessment of hydrocarbon biodegradation in hypersaline environments are presented in this review.