RESUMO
Enzymes of the archaea living in extreme environments are resistant to the challenging conditions. Lipase is among the important enzymes used in the industry and agriculture. In this study, the extracellular lipase from extremely halophilic archaeon Halolamina sp. was characterized for the first time. Optimum temperature for the enzyme activity was determined as 70oC, optimum pH was 7.0, and the optimum salt concentration was 3.6 M. Additionally, more than 70% of the enzyme activity was remained between pH 3.0-10.0 for 48 h as well as incubation of the enzyme at 70oC for 30 min increased its activity for 44%, and no activity loss was observed after incubation at 80oC. Also, presence of the metals increased the enzyme activity up to 88%. The enzyme was highly resistant to the organic solvents acetone, methanol, and DMSO while strong inhibition was caused by n-butanol. Among the detergents, the enzyme kept its activity substantially in the presence of SDS; however, other detergents caused inhibition of the enzyme activity. This characterization study showed that the lipase from the haloarchaeon Halolamina sp. is highly stable at the wide ranges of temperature and pH values as well as in the presence of diverse inhibitors. This enzyme is promising to be used in biotechnological applications.
Assuntos
Estabilidade Enzimática , Halobacteriales , Archaea , LipaseRESUMO
Handcrafted salted fish is marketed weekly in retail markets and public fairs in the Porto Velho city, Rondônia sate, Brazil. Knowing the microbiological quality of these products is essential for public health, given that such products are not subject to any quality control. The aimed of the study was to evaluate the hygienic-sanitary characteristics and the moisture content of pirarucu and salted and dried shrimp marketed in the Porto Velho city. Twenty samples were collected, 10 pirarucu and 10 shrimp, in February and June 2019. For microbiological analysis, surface plating was performed using acidified potato agar, for molds and yeasts, and PCA agar for halophilic bacterias. Moisture was determined by gravimetry, using an oven at 105° C. Results for molds and yeasts on pirarucu ranged <10 log CFU/g (absent) to 3 log CFU/g. For shrimp, values ranged <10 log CFU/g to 3.57 log CFU/g. For halophilic bacterias, contents ranged <10 log CFU/g to 6.30 log CFU/g in pirarucu samples and <10 log CFU/g to 6.97 log CFU/g in shrimp samples. The variation in moisture content ranged 36.99 to 54.31% for pirarucu, and 40.6 to 56.82% for shrimp. The results obtained may be related to poor hygienic conditions in processing, the lack of hygiene of utensils and places for handling, or even the quality of the raw material used.(AU)
Pescado salgado preparado artesanalmente é semanalmente comercializado em mercados varejistas e feiras públicas na cidade de Porto Velho, Rondônia. Conhecer a qualidade microbiológica desses produtos é essencial para a saúde pública, haja visto, que tais produtos não estão sujeitos a nenhum controle de qualidade. O objetivo do estudo foi avaliar as caracterís-ticas higiênico-sanitárias e o teor de umidade de pirarucus e camarões salgados e secos comercializados em Porto Velho. Foram coletadas 20 amostras, 10 de pirarucu e 10 de camarão, em fevereiro e junho de 2019. Para as análises microbiológicas foi realizado o plaqueamento em superfície usando-se ágar batata acidificado, para bolores e leveduras, e ágar PCA para as bacté-rias halofílicas. A umidade foi determinada por gravimetria, com uso de estufa a 105°C. Os resultados para bolores e leveduras em pirarucu variaram de <10 log UFC/g (ausente) a 3 log UFC/g. Para os camarões os valores variaram de <10 log UFC/g a 3,57 log UFC/g. Para as bactérias halofílicas os teores variaram de <10 log UFC/g a 6,30 log UFC/g nas amostras de pirarucu e de <10 log UFC/g a 6,97 log UFC/g nas amostras de camarão. A variação nos teores de umidade foi de 36,99 a 54,31% para o pirarucu, e de 40,6 a 56,82% para o camarão. Os resultados obtidos podem estar relacionados às más condições higiênicas no processamento, à falta de higiene dos utensílios e dos locais de manipulação, ou ainda, à qualidade da matéria-prima empregada.(AU)
Assuntos
Perciformes/microbiologia , Inspeção de Alimentos/métodos , Perfis Sanitários , Palaemonidae/microbiologia , Caça , Leveduras , Halobacteriales , FungosRESUMO
Salted and ripened fish foods are susceptible to cause histamine poisoning. The present study focuses on microbial histamine degradation from high salted fermented fishery products to deepen our understanding about this new and growing field of research. As a result of this first study related to salted-ripened anchovies (Engraulis anchoita), fifty seven moderate and extreme halophilic microbial isolates from salt and salted-ripened anchovy processes were characterized in terms of their phenotype and histamine-degrading capacity. Only 7%-4 isolates-were able to degrade histamine. None of the histamine-degrading isolates presented proteolytic and/or lipolytic activity. One of them designated A18 was chemotactic toward histamine, an interesting property not previously reported for that chemoattractant. However, the S18 and A18 isolates, genotypically identified as Halobacterium sp. and Halomonas sp. respectively, produced indole and/or H2S, both undesirable characteristics associated to off-flavors occurrence. On the other hand, A28 and S20, identified as Halovibrio sp. and Halobacterium sp. respectively, presented desirable properties, such as cytochrome oxidase and catalase activity, and non-production of H2S and indole. These strains also showed characteristics previously reported as dominant in the ripened stage. The results are promising, and A28 and S20 may have the desirable features to improve the anchovy salting-ripening process.
Assuntos
Halobacteriales , Halomonas , Histamina , Animais , Aquicultura , Peixes , Histamina/metabolismo , RNA Ribossômico 16S , Cloreto de SódioRESUMO
Halophilic bacteria are microorganisms that grow optimally in the presence of the very high concentration of sodium chloride. Halophiles are vital sources of various enzymes including hydrolases, which are very stable and catalytically highly efficient at high salt concentration and other extreme conditions such as high temperature, pH and presence of organic solvents. Several hydrolases such as amylases, proteases, and lipases have been obtained from halophilic bacteria and are commonly used for various industrial applications. We initiated a screening to isolate and characterize the halophilic bacteria from the Red Sea, which is one of the saltiest bodies of water in the world. Water and soil samples, collected from the Red Sea coast, Jeddah, Saudi Arabia, were screened for isolation of halophilic bacteria. Ten bacterial isolates were obtained, which were characterized by biochemical tests and 16S rRNA gene sequencing. Hydrolase producing bacteria among the isolates were screened by plate assay on starch and gelatin agar plates for amylase and protease, respectively. Two bacterial isolates i.e. Bacillus glycinifermentans S3 and Enterobacter cloacae W1were found to possess significant amylase and protease activity.
Bactérias halofílicas são microrganismos que crescem de maneira ideal na presença de uma concentração muito alta de cloreto de sódio. Halófilos são fontes vitais de várias enzimas, incluindo hidrolases, que são muito estáveis e cataliticamente altamente eficientes em alta concentração de sal e outras condições extremas, como alta temperatura, pH e presença de solventes orgânicos. Várias hidrolases como amilases, proteases e lipases foram obtidas a partir de bactérias halofílicas e são comumente usadas para várias aplicações industriais. Iniciamos uma triagem para isolar e caracterizar as bactérias halofílicas do Mar Vermelho, que é um dos corpos de água mais salgados do mundo. Amostras de água e solo, coletadas na costa do Mar Vermelho, Jeddah, na Arábia Saudita, foram examinadas quanto ao isolamento de bactérias halofílicas. Foram obtidos dez isolados bacterianos, caracterizados por testes bioquímicos e seqüenciamento do gene 16S rRNA. As bactérias produtoras de hidrolase entre os isolados foram triadas por ensaio em placa em placas de amido e ágar de gelatina para amilase e protease, respectivamente. Verificou-se que dois isolados bacterianos, isto é, Bacillus glycinifermentans S3 e Enterobacter cloacae W1, possuíam significativa atividade de amilase e protease.
Assuntos
Peptídeo Hidrolases , Halobacteriales , Salinidade , Amilases , HidrolasesRESUMO
Seasonal sampling was carried out at three Argentinian salterns, Salitral Negro (SN), Colorada Grande (CG) and Guatraché (G), to analyze abiotic parameters and microbial diversity and dynamics. Microbial assemblages were correlated to environmental factors by statistical analyses. Principal component analysis of the environmental data grouped SN and CG samples separately from G samples owing to G's higher pH values and sulfate concentration. Differences in microbial assemblages were also found. Many archaeal sequences belonged to uncultured members of Haloquadratum and Haloquadratum-related genera, with different environmental optima. Notably, nearly half of the archaeal sequences were affiliated to the recently described 'Candidatus Haloredividus' (phylum Nanohaloarchaeota), not previously detected in salt-saturated environments. Most bacterial sequences belonged to Salinibacter representatives, while sequences affiliated to the recently described genus Spiribacter were also found. Seasonal analysis showed at least 40% of the microbiota from the three salterns was prevalent through the year, indicating they are well adapted to environmental fluctuations. On the other hand, a minority of archaeal and bacterial sequences were found to be seasonally distributed. Five viral morphotypes and also eukaryal predators were detected, suggesting different mechanisms for controlling prokaryotic numbers. Notably, Guatraché was the saltern that harbored the highest virus-to-cell ratios reported to date for hypersaline environments.
Assuntos
Archaea/classificação , Bacteroidetes/classificação , Halobacteriales/classificação , Proteobactérias/classificação , Vírus/classificação , Archaea/genética , Archaea/isolamento & purificação , Argentina , Bacteroidetes/genética , Bacteroidetes/isolamento & purificação , Eletroforese em Gel de Campo Pulsado , Halobacteriales/genética , Microbiota/genética , Microscopia Eletrônica de Transmissão , Análise de Componente Principal , Proteobactérias/genética , Proteobactérias/isolamento & purificação , Estações do Ano , Água do Mar/microbiologia , Cloreto de Sódio , Vírus/genética , Vírus/isolamento & purificaçãoRESUMO
Halophiles are excellent sources of enzymes that are not only salt stable but also can withstand and carry out reactions efficiently under extreme conditions. The aim of the study was to isolate and study the diversity among halophilic bacteria producing enzymes of industrial value. Screening of halophiles from various saline habitats of India led to isolation of 108 halophilic bacteria producing industrially important hydrolases (amylases, lipases and proteases). Characterization of 21 potential isolates by morphological, biochemical and 16S rRNA gene analysis found them related to Marinobacter, Virgibacillus, Halobacillus, Geomicrobium, Chromohalobacter, Oceanobacillus, Bacillus, Halomonas and Staphylococcus genera. They belonged to moderately halophilic group of bacteria exhibiting salt requirement in the range of 3-20%. There is significant diversity among halophiles from saline habitats of India. Preliminary characterization of crude hydrolases established them to be active and stable under more than one extreme condition of high salt, pH, temperature and presence of organic solvents. It is concluded that these halophilic isolates are not only diverse in phylogeny but also in their enzyme characteristics. Their enzymes may be potentially useful for catalysis under harsh operational conditions encountered in industrial processes. The solvent stability among halophilic enzymes seems a generic novel feature making them potentially useful in non-aqueous enzymology.
Assuntos
Ativadores de Enzimas/análise , Biodiversidade , Halobacteriales/isolamento & purificação , Hidrolases/análise , Hidrolases/isolamento & purificação , Solventes/análise , Catálise , Microbiologia Ambiental , MétodosRESUMO
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
Microbiologia Ambiental , Halobacteriales/isolamento & purificação , Hidrocarbonetos/análise , Hidrocarbonetos/isolamento & purificação , Petróleo/análise , Solos Salitrosos , Metodologia como AssuntoRESUMO
We investigated the phylogenetic diversity and community structure of members of the halophilic Archaea (order Halobacteriales) in five distinct sediment habitats that experience various levels of salinity and salinity fluctuations (sediments from Great Salt Plains and Zodletone Spring in Oklahoma, mangrove tree sediments in Puerto Rico, sediment underneath salt heaps in a salt-processing plant, and sediments from the Great Salt Lake northern arm) using Halobacteriales-specific 16S rRNA gene primers. Extremely diverse Halobacteriales communities were encountered in all habitats, with 27 (Zodletone) to 37 (mangrove) different genera identified per sample, out of the currently described 38 Halobacteriales genera. With the exception of Zodletone Spring, where the prevalent geochemical conditions are extremely inhospitable to Halobacteriales survival, habitats with fluctuating salinity levels were more diverse than permanently saline habitats. Sequences affiliated with the recently described genera Halogranum, Halolamina, Haloplanus, Halosarcina, and Halorientalis, in addition to the genera Halorubrum, Haloferax, and Halobacterium, were among the most abundant and ubiquitous genera, suggesting a wide distribution of these poorly studied genera in saline sediments. The Halobacteriales sediment communities analyzed in this study were more diverse than and completely distinct from communities from typical hypersaline water bodies. Finally, sequences unaffiliated with currently described genera represented a small fraction of the total Halobacteriales communities, ranging between 2.5% (Zodletone) to 7.0% (mangrove and Great Salt Lake). However, these novel sequences were characterized by remarkably high levels of alpha and beta diversities, suggesting the presence of an enormous, yet-untapped supply of novel Halobacteriales genera within the rare biosphere of various saline ecosystems.
Assuntos
Biota , Sedimentos Geológicos/microbiologia , Halobacteriales/classificação , Halobacteriales/isolamento & purificação , Filogenia , Análise por Conglomerados , DNA Arqueal/química , DNA Arqueal/genética , DNA Ribossômico/química , DNA Ribossômico/genética , Ecossistema , Porto Rico , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Estados UnidosRESUMO
The present study deals with the isolation and characterization of the moderately halophilic-alkaliphilic bacteria from a saline habitat in western India. Eight different bacterial strains were isolated using enrichment techniques at 20 percent (w/v) NaCl and pH 10. The isolates exhibited diversity towards gram's reaction, colony and cell morphology. They were able to grow and produce alkaline protease over a broad range of NaCl, 5-20 percent (w/v) and pH, 8-10. None of the isolates could grow at pH 7, and one could not grow even at pH 8. Crude and partially purified proteases from strain S5 were subjected to characterization with reference to pH, salt stability and protein folding. Optimum protease activity and stability was recorded at 10 percent salt and pH 9-9.5. Denaturation kinetics of S5 alkaline protease along with a reference protease was studied at 8M urea followed by renaturation. The S5 alkaline protease could be partially renatured up to 32 percent of the original activity. Despite of the fact that all the 8 isolates were from the same site, they displayed significant diversity with respect to their salt requirement for growth and enzyme secretion. While the effect of pH was less demarcated on growth, the protease production was significantly affected. Isolate S5 produced substantial amount of halotolerant and alkaline protease. The activity and stability of the alkaline protease in a broader range of pH and salt would definitely make this enzyme an important candidate for various industrial applications.
O presente estudo relata o isolamento e caracterização de bactérias moderadamente halofilicas e alcalífilicas de um habitat salino no oeste da índia. Oito cepas diferentes de bactérias foram isoladas empregando técnicas de enriquecimento em NaCl a 20 por cento (p/v) e pH 10. As cepas apresentaram diversidade em relação à coloração de Gram e à morfologia das colônias e células. As cepas foram capazes de multiplicar e produzir protease alcalina em uma ampla faixa de concentração de NaCl (5 a 20 por cento) e pH (8 a 10). Nenhuma das cepas foi capaz de se multiplicar em pH 7, e uma não se multiplicou nem em pH 8.0. Proteases naturais e parcialmente purificadas da cepa S5 foram submetidas à caracterização com relação ao pH, estabilidade salina, e estrutura protéica. Atividade e estabilidade ótimas da protease foram obtidas com 10 por cento de sal e pH 9-9,5. A cinética de denaturação da protease de S5, juntamente com uma protease de referencia, foi avaliada com uréia 8M seguida de renaturação. A protease alcalina de S5 foi renaturada a 32 por cento da atividade original. Apesar de provenientes do mesmo local, as oito cepas mostraram grande diversidade em relação à exigência de sal para multiplicação e secreção enzimática. Enquanto o efeito do pH na multiplicação foi menos marcante, o efeito na produção de protease foi significativamente afetada. A cepa S5 produziu uma quantidade substancial de protease alcalina e halotolerante. A atividade e estabilidade da protease alcalina em uma faixa mais ampla de pH e sal tornam essa enzima uma importante candidata para diversas aplicações industriais.
Assuntos
Bactérias/isolamento & purificação , Secreções Corporais , Endopeptidases , Halobacteriales , Técnicas In Vitro , Solos Salitrosos , Métodos , Estudos de AmostragemRESUMO
Raman spectra have been obtained for extremophiles from several geological environments; selected examples have been taken from hot and cold deserts comprising psychrophiles, thermophiles and halophiles. The purpose of this study is the assessment of the effect of the wavelength of the laser excitation on the ability to determine unique information from the Raman spectra about the specificity of detection of biomolecules produced as a result of the survival strategies adopted by organisms in extreme terrestrial environments. It was concluded that whereas FT-Raman spectroscopy at 1064 nm gave good quality results the time required to record the data was relatively large compared with other wavelengths of excitation but that better access to the CH stretching region for organic molecules was given. Shorter wavelength excitation of biomolecules in the blue-green regions of the visible spectrum using a conventional dispersive spectrometer was more rapid but very dependent upon the type of chemical compound being studied; most relevant biomolecules fluoresced at these wavelengths but carotenoids exhibited a resonance effect which resulted in an improved detection capability. Minerals and geological materials, in contrast, were best studied at these visible wavelengths. In general, the best compromise system for the excitation of the Raman spectra of both geological and biological materials was provided using a 785 nm laser coupled with a dispersive spectrometer, especially for accessing the 1800-200 cm(-1) wavenumber shift region where much of the definitive analytical information resides. This work will have conclusions relevant to the use of miniaturised Raman spectrometers for the detection of biomolecules in extraterrestrial planetary exploration.
Assuntos
Biofilmes , Cianobactérias/química , Meio Ambiente , Halobacteriales/química , Líquens/química , Análise Espectral Raman/métodos , Regiões Antárticas , Regiões Árticas , Chile , Clima Frio , Clima DesérticoRESUMO
The term halophile is used for all those organisms belonging to hypersaline habitats; they constitute an interesting class of organisms able to compete successfully in salt water and to resist its denaturing effects. A wide diversity of microorganisms, prokaryotic and eukaryotic belong to this category. Halophile organisms have strategies allowing them not only to withstand osmotic stress, but also to function better in the presence of salt, in spite of maintaining high intracellular concentrations of salt, partly due to the synthesis of compatible solutes that allow them to balance their osmotic pressure. We describe the characteristics of some halophile organisms and D. hansenii (halophile yeast), that allow them to resist high concentrations of salt. The interest to know the great diversity microorganisms living in hypersaline habitats is growing, and has begun to be the center of recent investigations, since halophile organisms produce an wide variety of biomolecules that can be used for different applications. In this review we describe some mechanisms with which some halophile organisms count to resist the high concentration of salts, mainly NaCl.
Assuntos
Adaptação Fisiológica , Halobacteriales/fisiologia , Saccharomycetales/fisiologia , Animais , Proteínas de Bactérias/fisiologia , Biotecnologia/métodos , Clorófitas/fisiologia , Metabolismo Energético , Ativação Enzimática , Células Eucarióticas/efeitos dos fármacos , Células Eucarióticas/fisiologia , Proteínas Fúngicas/fisiologia , Regulação Bacteriana da Expressão Gênica , Regulação Fúngica da Expressão Gênica , Halobacteriales/efeitos dos fármacos , Líquido Intracelular/química , Lipídeos de Membrana/metabolismo , Proteínas Quinases Ativadas por Mitógeno/fisiologia , Concentração Osmolar , Pressão Osmótica , Fenômenos Fisiológicos Vegetais , Saccharomycetales/efeitos dos fármacos , Solução Salina Hipertônica/farmacologia , Solubilidade , Álcoois Açúcares/metabolismo , Transcrição GênicaRESUMO
An extremely halophilic Archaeon belonging to the order Halobacteriales was isolated from the solar salterns of Cabo Rojo, Puerto Rico. The organism, designated strain PR5T, is rod-shaped, non-motile and requires at least 12% (w/v) NaCl to grow. The strain is highly thermotolerant: its temperature optimum is 50 degrees C and growth is possible up to 60 degrees C. Polar lipid analysis revealed the presence of the bis-sulfated glycolipid S2-DGD-1 as sole glycolipid and the absence of the glycerol diether analogue of phosphatidylglycerosulfate. Both C20,C20 and C20,C25 core lipids are present. The G+C content of the DNA is 63.3 mol%. According to 16S rDNA sequence data, strain PR5T is closely related to the representatives of the genera Haloterrigena and Natrinema, but on the basis of its phenotypic properties, 16S rDNA sequence and DNA-DNA hybridization studies, strain PR5T cannot be assigned to any of the recognized species within these genera. On the basis of its polar lipid composition, the isolate has been assigned to the genus Haloterrigena. The creation of a new species, Haloterrigena thermotolerans, is therefore proposed to accommodate this isolate. The type strain is strain PR5T (= DSM 11552T = ATCC 700275T).