RESUMO
Dissimilatory arsenate-respiring prokaryotes (DARPs) are considered to be a key impetus of the reductive dissolution of solid-phase arsenic. However, little is known about the interaction between nitrate and DARPs so far. In this study, we showed that nitrate either inhibited or promoted the DARP population-catalyzed reductive mobilization of As in sediments. Metagenomic analysis of the microbial communities in the microcosms after seven days of As release assays suggested that microbes mainly consisted of: Type-I DARPs having potential to reduce NO3- into NO2- and Type-II DARPs having potential to reduce NO3- to NH4+. We further isolated two cultivable DARPs, Neobacillus sp. A01 and Paenibacillus sp. A02, which represent Type-I and -II DARPs, respectively. We observed that nitrate suppressed A01-mediated release of As(III) but promoted A02-mediated release of As(III). Furthermore, we demonstrated that this observation was due to the fact that nitrite, the end product of incomplete denitrification by Type-I DARPs, suppressed the arrA gene expression per cell and growth of all DARPs, whereas ammonium, the end product of dissimilatory nitrate reduction to ammonium (DNRA) by Type-II DARPs, enhanced the arrA gene expression per cell and significantly promoted the growth of all DARPs. These findings suggest that the actual effects of nitrate on DARP population-catalyzed reductive mobilization of arsenic, largely depend on the ratio of Type-I to Type-II DARPs in sediments.
Assuntos
Arsênio , Nitratos , Arseniatos , NitritosRESUMO
Many investigations suggest that dissimilatory arsenate-respiring prokaryotes (DARPs) play a key role in stimulating reductive mobilization of As from solid phase into groundwater, but it is not clear how environmental Mn(II) affects the DARPs-mediated reductive mobilization of arsenic. To resolve this issue, we collected soil samples from a realgar tailings-affected area. We found that there were diverse arsenate-respiratory reductase (arr) genes in the soils. The microbial communities had high arsenate-respiring activity, and were able to efficiently stimulate the reductive mobilization of As. Compared to the microcosms without Mn(II), addition of 10 mmol/L Mn(II) to the microcosms led to 23.99%-251.79% increases in the microbial mobilization of As, and led to 133.3%-239.2% increases in the abundances of arr genes. We further isolated a new cultivable DARP, Bacillus sp. F11, from the arsenic-contaminated soils. It completely reduced 1 mmol/L As(V) in 5 days under the optimal reaction conditions. We further found that it was able to efficiently catalyze the reductive mobilization and release of As from the solid phase; the addition of 2 mmol/L Mn(II) led to 98.49%-248.78% increases in the F11 cells-mediated reductive mobilization of As, and 70.6%-104.4% increases in the arr gene abundances. These data suggest that environmental Mn(II) markedly increased the DARPs-mediated reductive mobilization of As in arsenic-contaminated soils. This work provided a new insight into the close association between the biogeochemical cycles of arsenic and manganese.
Assuntos
Arsênio , Água Subterrânea , Arsênio/metabolismo , Arseniatos/metabolismo , SoloRESUMO
Arsenite (As(III)) was considered to be of great concern in acid mine drainage (AMD). A promising approach for cleaning up of arsenite from AMD is microbial oxidation of As(III) followed by adsorptions. However, there is virtually no research about the acidophilic bioreactor for As(III) oxidation so far. In this study, we formed a new biofilm bioreactor with a consortium of acidophilic As(III) oxidation bacteria. It is totally chemoautotrophic, with no need to add any carbon or other materials during the operations. It works well under pH 3.0-4.0, capable of oxidizing 1.0-20.0 mg/L As(III) in 3.0-4.5 h, respectively. A continuous operation of the bioreactor suggests that it is very stable and sustainable. Functional gene detection indicated that the biofilms possessed a unique diversity of As(III) oxidase genes. Taken together, this acidophilic bioreactor has great potential for industrial applications in the cleaning up of As(III) from AMD solution.
Assuntos
Arsenitos , Bactérias/genética , Biofilmes , Reatores Biológicos , OxirreduçãoRESUMO
The soils near the abandoned Shimen Realgar Mine are characterized by containing extremely high contents of total and soluble arsenic. To determine the microbial reactions and environmental factors affecting the mobilization and release of arsenic from soils phase into pore water, we collected 24 soil samples from the representative points around the abandoned Shimen Realgar Mine. They contained 8310.84â¯mg/kg total arsenic and 703.21â¯mg/kg soluble arsenic in average. The soluble arsenic in the soils shows significant positive and negative correlations with environmental SO42-/TOC/pH/PO43-, and Fe/Mn, respectively. We found that diverse dissimilatory As(V)-respiring prokaryotes (DARPs) and As(III)-oxidizing bacteria (AOB) exist in all the examined soil samples. The activities of DARPs led to 65-1275% increase of soluble As(III) in the examined soils after 21.0 days of anaerobic incubation, and the microbial dissolution and releases of arsenic show significant positive and negative correlations with the environmental pH/TN and NH4+/PO43-, respectively. In comparison, the activities of AOB led to 24-346% inhibition of the dissolved oxygen-mediated dissolution of arsenic in the soils, and the AOB-mediated releases of As(V) show significant positive and negative correlations with the environmental SO42- and pH/NH4+, respectively. The microbial communities of 24 samples contain 54 phyla of bacteria that show extremely high diversities. Total arsenic, TOC, NO3- and pH are the key environmental factors that indirectly controlled the mobilization and release of arsenic via influencing the structures of the microbial communities in the soils. This work gained new insights into the mechanism for how microbial communities catalyze the dissolution and releases of arsenic from the soils with extremely high contents of arsenic.
Assuntos
Arsênio/análise , Microbiologia do Solo , Poluentes do Solo/análise , Aerobiose , Anaerobiose , Bactérias/metabolismo , Concentração de Íons de Hidrogênio , Nitratos/análise , Solo/química , SolubilidadeRESUMO
Microbial arsenic (As) methylation plays important roles in the As biogeochemical cycle. However, little is known about the diversity and functions of As-methylating microorganisms from the tailings of a Realgar Mine, which is characterized as containing extremely high concentrations of As. To address this issue, we collected five samples (T1-T5) from the tailings of Shimen Realgar Mine. Microcosm assays without addition of exogenous As and carbon indicated that all the five samples possess significant As-methylating activities, producing 0.8-5.7 µg/L DMAsV, and 1.1-10.7 µg/L MMAsV with an exception of T3, from which MMAsV was not detectable after 14.0 days of incubation. In comparison, addition of 20.0 mM lactate to the microcosms significantly enhanced the activities of these samples; the produced DMAsV and MMAsV are 8.0-39.7 µg/L and 5.8-38.3 µg/L, respectively. The biogenic DMAsV shows significant positive correlations with the Fe concentrations and negative correlations with the total nitrogen concentrations in the environment. A total of 63 different arsM genes were identified from the five samples, which code for new or new-type ArsM proteins, suggesting that a unique diversity of As-methylating microbes are present in the environment. The microbial community structures of the samples were significantly shaped by the environmental total organic carbon, total As contents and NO3- contents. These data help to better understand the microorganisms-catalyzed As methylation occurred in the environment with extremely high contents of As.
Assuntos
Arsênio , Mineração , Microbiota , Microbiologia do SoloRESUMO
An arsenic-resistant strain, CB3T, was isolated from arsenic-rich aquifers at the Jianghan Plain in Hubei, China. Phylogenetic and biochemical analysis suggested that it should represent a new species of the genus Pseudaminobacter in the family Phyllobacteriaceae. The 16S rRNA gene of CB3T shared the highest sequence similarities to those of the type strains Pseudaminobacter defluvii THI 051T (97.8â% identity) and Pseudaminobacter salicylatoxidans BN12T (97.4â%). The DNA-DNA relatedness values of CB3T with respect to strains belonging to the genus Pseudaminobacter were less than 70â%. The fatty acid profile of CB3T consisted of C16â:â0, cyclo-C19â:â0ω8c and summed feature 8 (C18â:â1ω7c and/or C18â:â1ω6c) as major components. The major polar lipids were phosphatidylcholine, phosphatidylglycerol, phosphatidyldimethylethanolamine, phosphatidylmonomethylethanolamine, phosphatidylethanolamine and diphosphatidylglycerol. The DNA G+C content was 61.4 mol%. On the basis of phenotypic, chemotaxonomic and phylogenetic data, strain CB3T was distinct from previously described Pseudaminobacter species. Therefore, we propose that strain CB3T represents a novel species of the genus Pseudaminobacter, Pseudaminobacterarsenicus sp. nov., strain CB3T (=CCTCC AB2016116T=KCTC 52625T) is designated as the type strain.
Assuntos
Arsênio , Água Subterrânea/microbiologia , Phyllobacteriaceae/classificação , Filogenia , Técnicas de Tipagem Bacteriana , Composição de Bases , China , DNA Bacteriano/genética , Ácidos Graxos/química , Hibridização de Ácido Nucleico , Fosfolipídeos/química , Phyllobacteriaceae/isolamento & purificação , RNA Ribossômico 16S/genética , Análise de Sequência de DNARESUMO
It was well established that microbial communities are the major drive for the formation of arsenic-contaminated groundwater. However, it remains to be elucidated for how nitrate/nitrite affects the microorganisms-catalyzed dissolution and reduction of arsenic. To address this issue, we collected soil samples containing high-contents of arsenic from the Shimen Realgar Mine area. Microcosm assay indicated that addition of nitrate/nitrite significantly inhibited the dissolution, reduction and release of As and Fe caused by the biological catalysis of microbial communities in the soils, meanwhile nitrate/nitrite was reduced into N2. To further investigate the molecular mechanism of this finding, we used a representative dissimilatory arsenate-respiring strain Shewanella sp. GL90 from the soils to perform the arsenic release assay. GL90 can efficiently catalyze the reductive dissolution, and promote the release of As and Fe in soils. It is interesting to see that the addition of nitrate/nitrite to the soils led to marked decreases in the GL90-mediated dissolution of As and Fe in the soils. Moreover, we found that this finding was attributed to that nitrate/nitrite significantly inhibited the transcription of the gene of the respiratory arsenate reductase protein in GL90 cells. This work provided new insights into the mechanisms for the coupling of As, N and Fe geochemical cycles in arsenic-rich soils, and for how environmental factors affect As concentration in groundwater.
Assuntos
Arsênio/metabolismo , Bactérias/metabolismo , Água Subterrânea/química , Ferro/metabolismo , Nitratos/análise , Nitritos/análise , Poluentes do Solo/metabolismo , China , Oxirredução , Microbiologia do Solo , Poluentes do Solo/análise , SolubilidadeRESUMO
Arsenite-oxidizing bacteria (AOB) play a key role in the biogeochemical cycle of arsenic in the environment, and are used for the bioremediation of As contaminated groundwater; however, it is not yet known about how arsenic affects biofilm formations of AOB, and how biofilm formations affect bacterial arsenite-oxidizing activities. To address these issues, we isolated seven novel AOB strains from the arsenic-contaminated soils. They can completely oxidize 1.0â¯mM As(III) in 22-60â¯h. Their arsenite oxidase sequences show 43-99% identities to those of other known AOB. Strains Cug1, Cug2, Cug3, Cug4, and Cug6 are able to form biofilms with thickness of 15-95⯵m, whereas Cug8 and Cug9 cannot form biofilms. It is interesting to see that arsenite inhibited the biofilm formations of heterotrophic AOB strains, but promoted the biofilm formations of autotrophic strains in a concentration-dependent manner. The arsenite-oxidizing rates of Cug1 and Cug4 biofilms are 31.6% and 27.6% lower than those of their suspension cultures, whereas the biofilm activities of other strains are similar to those of their suspension cultures. The biofilm formation significantly promoted the bacterial resistance to arsenic. This work is the first report on the complex correlations among environmental arsenic, bacterial biofilm formations and bacterial arsenite-oxidizing activities. The data highlight the diverse lifestyle of different AOB under arsenic stress, and provide essential knowledge for the screening of efficient AOB strains used for constructions of bioreactors.
Assuntos
Arsenitos/metabolismo , Bactérias/metabolismo , Biofilmes/efeitos dos fármacos , Poluentes do Solo/metabolismo , Arsênio/metabolismo , Arsenitos/toxicidade , Processos Autotróficos , Bactérias/efeitos dos fármacos , Bactérias/enzimologia , Bactérias/isolamento & purificação , Fenômenos Fisiológicos Bacterianos , Biodegradação Ambiental , Processos Heterotróficos , Oxirredução , Oxirredutases/genética , Microbiologia do Solo , Poluentes do Solo/toxicidadeRESUMO
The paddy soils in some areas in Jianghan Plain were severely contaminated by arsenic. However, little is known about the activity and diversity of the dissimilatory arsenate-respiring prokaryotes (DARPs) in the paddy soils, and the effects of sulfate on the microbial mobilization and release of arsenic from soils into solution. To address this issue, we collected arsenic-rich soils from the depths of 1.6 and 4.6 m in a paddy region in the Xiantao city, Hubei Province, China. Microcosm assays indicated that all of the soils have significant arsenate-respiring activities using lactate, pyruvate or acetate as the sole electron donor. Functional gene cloning and analysis suggest that there are diverse DARPs in the indigenous microbial communities of the soils. They efficiently promoted the mobilization, reduction and release of arsenic and iron from soils under anaerobic conditions. Remarkably, when sulfate was amended into the microcosms, the microorganisms-catalyzed reduction and release of arsenic and iron were significantly increased. We further found that sulfate significantly enhanced the arsenate-respiring reductase gene abundances in the soils. Taken together, a diversity of DARPs in the paddy soils significantly catalyzed the dissolution, reduction and release of arsenic and iron from insoluble phase into solution, and the presence of sulfate significantly increased the microbial reactions.
Assuntos
Arsênio/metabolismo , Microbiologia do Solo , Poluentes do Solo/metabolismo , Poluentes Químicos da Água/metabolismo , Arseniato Redutases/metabolismo , Arseniatos/metabolismo , China , Água Subterrânea/química , Solo/química , Sulfatos/metabolismoRESUMO
Highly acidic peptides with no disulfide bridges are widely present in the scorpion venoms; however, none of them has been functionally characterized so far. Here, we cloned the full-length cDNA of a short-chain highly acidic peptide (referred to as HAP-1) from a cDNA library made from the venom glands of the Chinese scorpion Mesobuthus martensii Karsch. HAP-1 contains 19 amino acid residues with a predicted IP value of 4.25. Acidic amino residues account for 33.3% of the total residues in the molecule of HAP-1. HAP-1 shows 76â»98% identities to some scorpion venom peptides that have not yet been functionally characterized. Secondary structure prediction showed that HAP-1 contains a beta-sheet region (residues 9â»17), and two coiled coil regions (residues 1â»8 and 18â»19) located at the N-terminal and C-terminal regions of the peptide, respectively. Antimicrobial assay showed that HAP-1 does not have any effect on the growth of the bacterium Staphylococcus aureus AB94004. However, it potently inhibits the antimicrobial activity of a 13-mer peptide from M. martensii Karsch against Staphylococcus aureus AB94004. This finding is the first characterization of the function of such highly acidic peptides from scorpions.
Assuntos
Anti-Infecciosos/química , Anti-Infecciosos/farmacologia , Venenos de Escorpião/química , Escorpiões/química , Animais , Peptídeos/química , Peptídeos/farmacologia , Staphylococcus aureus/efeitos dos fármacosRESUMO
Hot Springs have unique geochemical features. Microorganisms-mediated arsenite oxidation is one of the major biogeochemical processes occurred in some hot springs. This study aimed to understand the diversities of genes and microorganisms involved in arsenite oxidation from the outlet of an untraversed hot spring located at an altitude of 4226 m. Microcosm assay indicated that the microbial community from the hot spring was able to efficiently oxidize As(III) using glucose, lactic acid, yeast extract or sodium bicarbonate as the sole carbon source. The microbial community contained 7 phyla of microorganisms, of which Proteobacteria and Firmicutes are largely dominant; this composition is unique and differs significantly from those of other described hot springs. Twenty one novel arsenite oxidase genes were identified from the samples, which are affiliated with the arsenite oxidase families of α-Proteobacteria, ß-Proteobacteria or Archaea; this highlights the high diversity of the arsenite-oxidizing microorganisms from the hot spring. A cultivable arsenite-oxidizer Chelatococcu sp. GHS311 was also isolated from the sample using enrichment technique. It can completely convert 75.0 mg/L As(III) into As(V) in 18 days at 45 °C. The arsenite oxidase of GHS311 shares the maximal sequence identity (84.7%) to that of Hydrogenophaga sp. CL3, a non-thermotolerant bacterium. At the temperature lower than 30 °C or higher than 65 °C, the growth of this strain was completely inhibited. These data help us to better understand the diversity and functional features of the thermophilic arsenite-oxidizing microorganisms from hot springs.
Assuntos
Arsenitos/metabolismo , Termotolerância/genética , Microbiologia da Água , Poluentes Químicos da Água/metabolismo , Archaea/genética , Arsenitos/análise , Sedimentos Geológicos/química , Fontes Termais , Concentração de Íons de Hidrogênio , Oxirredução , Poluentes Químicos da Água/análiseRESUMO
The tailings of the Shimen realgar mine have unique geochemical features. Arsenite oxidation is one of the major biogeochemical processes that occurs in the tailings. However, little is known about the functional and molecular aspects of the microbial community involved in arsenite oxidation. Here, we fully explored the functional and molecular features of the microbial communities from the tailings of the Shimen realgar mine. We collected six samples of tailings from sites A, B, C, D, E, and F. Microcosm assays indicated that all of the six sites contain both chemoautotrophic and heterotrophic arsenite-oxidizing microorganisms; their activities differed considerably from each other. The microbial arsenite-oxidizing activities show a positive correlation with soluble arsenic concentrations. The microbial communities of the six sites contain 40 phyla of bacteria and 2 phyla of archaea that show extremely high diversity. Soluble arsenic, sulfate, pH, and total organic carbon (TOC) are the key environmental factors that shape the microbial communities. We further identified 114 unique arsenite oxidase genes from the samples; all of them code for new or new-type arsenite oxidases. We also isolated 10 novel arsenite oxidizers from the samples, of which 4 are chemoautotrophic and 6 are heterotrophic. These data highlight the unique diversities of the arsenite-oxidizing microorganisms and their oxidase genes from the tailings of the Shimen realgar mine. To the best of our knowledge, this is the first report describing the functional and molecular features of microbial communities from the tailings of a realgar mine. IMPORTANCE: This study focused on the functional and molecular characterizations of microbial communities from the tailings of the Shimen realgar mine. We fully explored, for the first time, the arsenite-oxidizing activities and the functional gene diversities of microorganisms from the tailings, as well as the correlation of the microbial activities/diversities with environmental factors. The findings of this study help us to better understand the diversities of the arsenite-oxidizing bacteria and the geochemical cycle of arsenic in the tailings of the Shimen realgar mine and gain insights into the microbial mechanisms by which the secondary minerals of the tailings were formed. This work also offers a set of unique arsenite-oxidizing bacteria for basic research of the molecular regulation of arsenite oxidation in bacterial cells and for the environmentally friendly bioremediation of arsenic-contaminated groundwater.
Assuntos
Archaea/genética , Archaea/metabolismo , Arsenitos/metabolismo , Bactérias/genética , Bactérias/metabolismo , Variação Genética , Archaea/classificação , Archaea/isolamento & purificação , Proteínas Arqueais/genética , Proteínas Arqueais/metabolismo , Bactérias/classificação , Bactérias/isolamento & purificação , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Biodegradação Ambiental , Mineração , Oxirredução , Oxirredutases/genética , Oxirredutases/metabolismo , FilogeniaRESUMO
A Gram-stain-negative, rod-shaped bacterium that formed yellow and viscous colonies was isolated from arsenic-contaminated soil of the Jianghan plain, Hubei Province, China, and it was designated 26-35T. This strain was capable of resisting arsenate and arsenite with MICs of 40 and 20 mM, respectively. The 16S rRNA gene of the novel isolate displayed 96.7-94.2 % sequence similarities to those of other known species of the genus Luteimonas. The respiratory quinone was ubiquinone-8 (Q-8). The DNA G+C content was 71.4 mol%. The predominant cellular fatty acids were iso-C15 : 0, iso-C16 : 0, iso-C17 : 0, iso-C11 : 0, iso-C11 : 0 3-OH and iso-C17 : 1ω9c. The major polar lipids were diphosphatidylglycerol, phosphatidylethanolamine and phosphatidylglycerol. Phylogenetic and physiological analysis indicated that the isolate represents a novel species of the genus Luteimonas, for which the name Luteimonas arsenica sp. nov. is proposed. The type strain is 26-35T (=KCTC 42824T=CCTCC AB 2014326T).
Assuntos
Arsênio/química , Filogenia , Microbiologia do Solo , Poluentes do Solo/química , Xanthomonadaceae/classificação , Técnicas de Tipagem Bacteriana , Composição de Bases , China , DNA Bacteriano/genética , Ácidos Graxos/química , Fosfolipídeos/química , Pigmentação , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Ubiquinona/química , Xanthomonadaceae/genética , Xanthomonadaceae/isolamento & purificaçãoRESUMO
A novel arsenic-resistant bacterium, designated 42-50T, was isolated from the high-arsenic sediment of Jianghan Plain, Hubei Province, China. Phylogenetic and biochemical analysis indicated that this bacterium represents the first species of a novel genus belonging to the family Hyphomicrobiaceae. The 16S rRNA gene of strain 42-50T shares 96.3-94.2, 96.3, 96.2 and 94.9-93.8 % sequence identities to those of species from the genera Devosia, Youhaiella, Paradevosia and Pelagibacterium, respectively. The major cellular fatty acids are C16 : 0, C18 : 0, C18 : 1ω7c 11-methyl and summed feature 8 (comprising C18 : 1ω7c and C18 : 1ω6c). The predominant polar lipids are diphosphatidylglycerol, phosphatidylglycerol and two unidentified glycolipids. The predominant respiratory quinone is ubiquinone-10 (Q-10). The DNA G+C content of strain 42-50T is 73.7 mol%. The distinct phylogenetic lineage and unique cellular fatty acids suggest that strain 42-50T represents a novel species of a new genus affiliated with the family Hyphomicrobiaceae, for which the name Arsenicitalea aurantiaca gen. nov., sp. nov. is proposed. The type strain is 42-50T (=CCTCC AB 2014325T=KCTC 42825T).
Assuntos
Arsênio , Sedimentos Geológicos/microbiologia , Hyphomicrobiaceae/classificação , Filogenia , Técnicas de Tipagem Bacteriana , Composição de Bases , China , DNA Bacteriano/genética , Ácidos Graxos/química , Glicolipídeos/química , Hyphomicrobiaceae/genética , Hyphomicrobiaceae/isolamento & purificação , Fosfatidilgliceróis/química , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Ubiquinona/químicaRESUMO
Antibiotic-resistant bacteria, such as methicillin-resistant Staphylococcus aureus and vancomycin-resistant Enterococcus, pose serious threat to human health. The outbreak of antibiotic-resistant pathogens in recent years emphasizes once again the urgent need for the development of new antimicrobial agents. Here, we discovered a novel antimicrobial peptide from the scorpion Opistophthalmus glabrifrons, which was referred to as Opisin. Opisin consists of 19 amino acid residues without disulfide bridges. It is a cationic, amphipathic, and α-helical molecule. Protein sequence homology search revealed that Opisin shares 42.1-5.3% sequence identities to the 17/18-mer antimicrobial peptides from scorpions. Antimicrobial assay showed that Opisin is able to potently inhibit the growth of the tested Gram-positive bacteria with the minimal inhibitory concentration (MIC) values of 4.0-10.0 µM; in contrast, it possesses much lower activity against the tested Gram-negative bacteria and a fungus. It is interesting to see that Opisin is able to strongly inhibit the growth of methicillin- and vancomycin-resistant pathogens with the MICs ranging from 2.0 to 4.0 µM and from 4.0 to 6.0 µM, respectively. We found that at a concentration of 5 × MIC, Opisin completely killed all the cultured methicillin-resistant Staphylococcus aureus. These results suggest that Opisin is a promising therapeutic candidate for the treatment of the antibiotic-resistant bacterial infections.
Assuntos
Antibacterianos/química , Antibacterianos/farmacologia , Anti-Infecciosos/farmacologia , Cisteína/química , Escorpiões/microbiologia , Animais , Farmacorresistência Bacteriana , Bactérias Gram-Negativas/efeitos dos fármacos , Bactérias Gram-Positivas/efeitos dos fármacos , Testes de Sensibilidade Microbiana , Dados de Sequência Molecular , Venenos de Escorpião/químicaRESUMO
A novel type of venom peptide with six disulfide bridges, referred to as Androcin, was identified from the scorpion Androctonus bicolor using a cDNA library strategy. The amino acid sequence of Androncin displays little identity to other known peptides from scorpions. We found that the genomic sequence of Androcin consists of three exons interrupted by two introns that are localized in the signal sequence and mature peptide encoding regions, inserted in phase 2 and phase 0, respectively. This genomic organization is unique among those of the cysteine-rich peptides from scorpions described so far. The primary and secondary structures of Androcin are homologous to those of the N-terminal domains of insulin-like growth factor-binding proteins; this suggests that Androcin may block the normal function of IGFs. Toxicological analysis using the recombinant Androcin peptide revealed that Androcin is able to induce severe akinesia and anxiety-like symptoms in mice. Androcin is a novel mammalian toxin with six disulfide bridges that provides the scorpion with another tool to subdue animals.
Assuntos
Ansiedade/induzido quimicamente , Cisteína/metabolismo , Aprendizagem em Labirinto/efeitos dos fármacos , Fragmentos de Peptídeos/toxicidade , Proteínas Recombinantes/metabolismo , Venenos de Escorpião/toxicidade , Escorpiões/metabolismo , Sequência de Aminoácidos , Animais , Sequência de Bases , Clonagem Molecular , Biologia Computacional , Éxons/genética , Feminino , Biblioteca Gênica , Injeções Intraperitoneais , Íntrons/genética , Camundongos , Camundongos Endogâmicos BALB C , Dados de Sequência Molecular , Fragmentos de Peptídeos/administração & dosagem , Fragmentos de Peptídeos/genética , Filogenia , Proteínas Recombinantes/genética , Venenos de Escorpião/administração & dosagem , Venenos de Escorpião/genética , Escorpiões/genética , Homologia de Sequência de AminoácidosRESUMO
A novel bacterial strain Z(T) was isolated from the high-arsenic sediment in Jianghan Plain, China. The strain was Gram-staining-negative, rod-shaped and formed yellow colonies. This bacterium is capable of tolerating arsenate and arsenite, with MICs of 40 mM and 20 mM, respectively. The strain also possesses catalase and does not produce oxidase. The nucleotide sequence of the 16S rRNA gene of the isolate showed the highest similarity (96.9%) to that of the type strain of Flavobacterium soli. On the basis of the 16S rRNA gene sequence analysis and the phenotypic properties of strain Z(T), it was assigned to the genus Flavobacterium. The major respiratory menaquinone was MK-6 and the predominant fatty acids were iso-C15:0, summed feature 3 (containing C16:1ω6c and/or C16:1ω7c) and iso-C15:1G. The major polar lipids were phosphatidylethanolamine, three uncharacterized aminophospholipids and four unidentified phospholipids. The DNA G+C content was 32.1 mol%. Based on the phenotypic and genotypic data presented in this article, it can be concluded that this isolate represents a novel species of the genus Flavobacterium, for which the name Flavobacterium arsenatis sp. nov. is proposed. The type strain is Z(T) ( = CCTCC AB 2013048(T) = KCTC 32397(T)).
Assuntos
Arsênio/química , Flavobacterium/classificação , Sedimentos Geológicos/microbiologia , Filogenia , Técnicas de Tipagem Bacteriana , Composição de Bases , China , DNA Bacteriano/genética , Ácidos Graxos/química , Flavobacterium/genética , Flavobacterium/isolamento & purificação , Dados de Sequência Molecular , Fosfolipídeos/química , Pigmentação , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Vitamina K 2/análogos & derivados , Vitamina K 2/química , Poluentes Químicos da Água/químicaRESUMO
A Gram-staining-negative, rod-shaped and pink bacterium was isolated from the soil of a Populus euphratica forest in the Taklamakan desert, Xinjiang, China. It was designated strain H9X(T). A 16S rRNA gene sequence homology search indicated that the isolate was most closely related to the family Cytophagaceae. The 16S rRNA gene of strain H9X(T) displayed 94.2-96.3â% sequence identities to those of type strains of other species of the genus Pontibacter. It only possessed menaquinone-7. The major cellular fatty acids of the novel isolate were iso-C15â:â0, C16â:â1ω5c summed feature 3 (containing C16â:â1ω6c and/or C16â:â1ω7c) and summed feature 4 (comprising anteiso-C17â:â1 B and/or iso-C17â:â1 I). The major polar lipids were phosphatidylethanolamine, one unknown aminophospholipid, one unknown glycophospholipid and several unknown phospholipids. The DNA G+C content of this bacterium was 55.2 mol%. Based on the phenotypic and genotypic data presented, it can be concluded that this isolate represents a novel species of the genus Pontibacter, for which the name Pontibacter yuliensis sp. nov. is proposed. The type strain is H9X(T) (â=âCCTCC AB 2013047(T)â=âKCTC 32396(T)).
Assuntos
Cytophagaceae/classificação , Clima Desértico , Filogenia , Populus/microbiologia , Microbiologia do Solo , Técnicas de Tipagem Bacteriana , Composição de Bases , China , Cytophagaceae/genética , Cytophagaceae/isolamento & purificação , DNA Bacteriano/genética , Ácidos Graxos/química , Dados de Sequência Molecular , Fosfolipídeos/química , Pigmentação , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Árvores/microbiologia , Vitamina K 2/análogos & derivados , Vitamina K 2/químicaRESUMO
Radioactive wastes always contain radioactive substances and a lot of Pb compound and other heavy metals, which severely contaminate soils and groundwater. Thus, search for radiation-resistant microorganisms that are capable of sequestering Pb contaminants from the contaminated sites is urgently needed. However, very few such microorganisms have been found so far. In the present study, we discovered a novel Gram-negative bacterium from the arid Taklamakan desert, which can strongly resist both radiation and Pb(2+). Phylogenetic and phenotypic analysis indicated that this bacterial strain is closely affiliated with Microvirga aerilata, and was thus referred to as Microvirga aerilata LM (=CCTCC AB 208311). We found that M. aerilata LM can effectively accumulate Pb and form intracellular precipitations. It also keeps similar ability to remove Pb(2+) under radioactive stress. Our data suggest that M. aerilata LM may offer an effective and eco-friendly in situ approach to remove soluble Pb contaminants from radioactive wastes.
Assuntos
Bactérias/classificação , Chumbo/metabolismo , Resíduos Radioativos/análise , Poluentes do Solo/metabolismo , Bactérias/genética , Bactérias/isolamento & purificação , Bactérias/metabolismo , Biodegradação Ambiental , Chumbo/análise , Poluentes do Solo/análiseRESUMO
Sb(III) and As(III) share similar chemical features and coexist in the environment. However, their oxidase enzymes have completely different sequences and structures. This raises an intriguing question: Could Sb(III)-oxidizing prokaryotes (SOPs) also oxidize As(III), and vice versa? Regarding this issue, previous investigations have yielded unclear, incorrect and even conflicting data. This work aims to address this matter. First, we prepared an enriched population of SOPs that comprises 55 different AnoA genes, lacking AioAB and ArxAB genes. We found that these SOPs can oxidize both Sb(III) and As(III) with comparable capabilities. To further confirm this finding, we isolated three cultivable SOP strains that have AnoA gene, but lack AioAB and ArxAB genes. We observed that they also oxidize both Sb(III) and As(III) under both anaerobic and aerobic conditions. Secondly, we obtained an enriched population of As(III)-oxidizing prokaryotes (AOPs) from As-contaminated soils, which comprises 69 different AioA genes, lacking AnoA gene. We observed that the AOP population has significant As(III)-oxidizing activities, but lack detectable Sb(III)-oxidizing activities under both aerobic and anaerobic conditions. Therefore, we convincingly show that SOPs can oxidize As(III), but AOPs cannot oxidize Sb(III). These findings clarify the previous ambiguities, confusion, errors or contradictions regarding how SOPs and AOPs oxidize each other's substrate.