RESUMO
Sideroxydans lithotrophicus ES-1 grows autotrophically either by Fe(II) oxidation or by thiosulfate oxidation, in contrast to most other isolates of neutrophilic Fe(II)-oxidizing bacteria (FeOB). This provides a unique opportunity to explore the physiology of a facultative FeOB and constrain the genes specific to Fe(II) oxidation. We compared the growth of S. lithotrophicus ES-1 on Fe(II), thiosulfate, and both substrates together. While initial growth rates were similar, thiosulfate-grown cultures had higher yield with or without Fe(II) present, which may give ES-1 an advantage over obligate FeOB. To investigate the Fe(II) and S oxidation pathways, we conducted transcriptomics experiments, validated with reverse transcription-quantitative PCR (RT-qPCR). We explored the long-term gene expression response at different growth phases (over days to a week) and expression changes during a short-term switch from thiosulfate to Fe(II) (90 min). The dsr and sox sulfur oxidation genes were upregulated in thiosulfate cultures. The Fe(II) oxidase gene cyc2 was among the top expressed genes during both Fe(II) and thiosulfate oxidation, and addition of Fe(II) to thiosulfate-grown cells caused an increase in cyc2 expression. These results support the role of Cyc2 as the Fe(II) oxidase and suggest that ES-1 maintains readiness to oxidize Fe(II), even in the absence of Fe(II). We used gene expression profiles to further constrain the ES-1 Fe(II) oxidation pathway. Notably, among the most highly upregulated genes during Fe(II) oxidation were genes for alternative complex III, reverse electron transport, and carbon fixation. This implies a direct connection between Fe(II) oxidation and carbon fixation, suggesting that CO2 is an important electron sink for Fe(II) oxidation. IMPORTANCE Neutrophilic FeOB are increasingly observed in various environments, but knowledge of their ecophysiology and Fe(II) oxidation mechanisms is still relatively limited. Sideroxydans isolates are widely observed in aquifers, wetlands, and sediments, and genome analysis suggests metabolic flexibility contributes to their success. The type strain ES-1 is unusual among neutrophilic FeOB isolates, as it can grow on either Fe(II) or a non-Fe(II) substrate, thiosulfate. Almost all our knowledge of neutrophilic Fe(II) oxidation pathways comes from genome analyses, with some work on metatranscriptomes. This study used culture-based experiments to test the genes specific to Fe(II) oxidation in a facultative FeOB and refine our model of the Fe(II) oxidation pathway. We gained insight into how facultative FeOB like ES-1 connect Fe, S, and C biogeochemical cycling in the environment and suggest a multigene indicator would improve understanding of Fe(II) oxidation activity in environments with facultative FeOB.
Assuntos
Transcrição Reversa , Transcriptoma , Compostos Ferrosos/metabolismo , Gallionellaceae , Oxirredução , Reação em Cadeia da PolimeraseRESUMO
Fe(II) clays are common across many environments, making them a potentially significant microbial substrate, yet clays are not well established as an electron donor. Therefore, we explored whether Fe(II)-smectite supports the growth of Sideroxydans lithotrophicus ES-1, a microaerophilic Fe(II)-oxidizing bacterium (FeOB), using synthesized trioctahedral Fe(II)-smectite and 2% oxygen. S. lithotrophicus grew substantially and can oxidize Fe(II)-smectite to a higher extent than abiotic oxidation, based on X-ray near-edge spectroscopy (XANES). Sequential extraction showed that edge-Fe(II) is oxidized before interior-Fe(II) in both biotic and abiotic experiments. The resulting Fe(III) remains in smectite, as secondary minerals were not detected in biotic and abiotic oxidation products by XANES and Mössbauer spectroscopy. To determine the genes involved, we compared S. lithotrophicus grown on smectite versus Fe(II)-citrate using reverse-transcription quantitative PCR and found that cyc2 genes were highly expressed on both substrates, while mtoA was upregulated on smectite. Proteomics confirmed that Mto proteins were only expressed on smectite, indicating that ES-1 uses the Mto pathway to access solid Fe(II). We integrate our results into a biochemical and mineralogical model of microbial smectite oxidation. This work increases the known substrates for FeOB growth and expands the mechanisms of Fe(II)-smectite alteration in the environment.
Assuntos
Compostos Ferrosos , Ferro , Ferro/química , Argila , Oxirredução , Compostos Ferrosos/metabolismoRESUMO
Organic ligands are widely distributed and can affect microbially driven Fe biogeochemical cycles, but effects on microbial iron oxidation have not been well quantified. Our work used a model microaerophilic Fe(II)-oxidizing bacterium Sideroxydans lithotrophicus ES-1 to quantify biotic Fe(II) oxidation rates in the presence of organic ligands at 0.02 atm O2 and pH 6.0. We used two common Fe chelators with different binding strengths: citrate (log KFe(II)-citrate = 3.20) and nitrilotriacetic acid (NTA) (log KFe(II)-NTA = 8.09) and two standard humic substances, Pahokee peat humic acid (PPHA) and Suwannee River fulvic acid (SRFA). Our results provide rate constants for biotic and abiotic Fe(II) oxidation over different ligand concentrations and furthermore demonstrate that various models and natural iron-binding ligands each have distinct effects on abiotic versus biotic Fe(II) oxidation rates. We show that NTA accelerates abiotic oxidation and citrate has negligible effects, making it a better laboratory chelator. The humic substances only affect biotic Fe(II) oxidation, via a combination of chelation and electron transfer. PPHA accelerates biotic Fe(II) oxidation, while SRFA decelerates or accelerates the rate depending on concentration. The specific nature of organic-Fe microbe interactions may play key roles in environmental Fe(II) oxidation, which have cascading influences on cycling of nutrients and contaminants that associate with Fe oxide minerals.
Assuntos
Compostos Férricos , Compostos Ferrosos , Gallionellaceae , Ligantes , OxirreduçãoRESUMO
Quantum dots (QDs) are recognized as the excellent fluorescence and photochemical materials to be applied in bioimaging, biomedical, and solar cell fields. Biosynthesized QDs (bio-QDs) have attracted attention due to their simple, eco-friendly, and excellent biocompatible traits. Moreover, bio-QDs could not be replaced by chemically fabricated QDs in many fields. Bio-QDs synthesized by different microorganisms have diverse characteristics. In this work, the biosynthesis of QDs by Tetrahymena pyriformis, a typical protozoa in aquatic environments, was achieved for the first time. The synthesized materials by T. pyriformis emitted yellow fluorescence and had an average diameter of 8.27 ± 0.77 nm. Spectral characterization results demonstrated that the synthesized QDs were CdS1-XSeX. Meanwhile, the fluorescence intensities of the synthesized bio-QDs showed a linear relationship with Cd2+ dosage ranging from 20 to 80 µM. The fluorescence enhancement of the synthesized QDs was highly selective to Cd2+ compared to other metal ions. The bio-QDs were demonstrated to have a great potential to be applied for Cd2+ detection. This work provides valuable information about the transformation of heavy metal ions in protozoan and is useful to accelerate the applications of the synthesized QDs.
Assuntos
Cádmio/análise , Engenharia Metabólica/métodos , Redes e Vias Metabólicas/genética , Pontos Quânticos/metabolismo , Tetrahymena pyriformis/metabolismo , Cátions Bivalentes/análise , Fluorescência , Química Verde/métodos , Metais Pesados/análise , Pontos Quânticos/química , Análise Espectral , Tetrahymena pyriformis/genéticaRESUMO
Biofabrication of nanomaterials is currently constrained by a low production efficiency and poor controllability on product quality compared to chemical synthetic routes. In this work, we show an attractive new biosynthesis system to break these limitations. A directed production of selenium-containing nanoparticles in Shewanella oneidensis MR-1 cells, with fine-tuned composition and subcellular synthetic location, was achieved by modifying the extracellular electron transfer chain. By taking advantage of its untapped intracellular detoxification and synthetic power, we obtained high-purity, uniform-sized cadmium selenide nanoparticles in the cytoplasm, with the production rates and fluorescent intensities far exceeding the state-of-the-art biosystems. These findings may fundamentally change our perception of nanomaterial biosynthesis process and lead to the development of fine-controllable nanoparticles biosynthesis technologies.
RESUMO
Biosynthesis offers opportunities for cost-effective and sustainable production of semiconductor quantum dots (QDs), but is currently restricted by poor controllability on the synthesis process, resulting from limited knowledge on the assembly mechanisms and the lack of effective control strategies. In this work, we provide molecular-level insights into the formation mechanism of biogenic QDs (Bio-QDs) and its connection with the cellular substrate metabolism in Escherichia coli. Strengthening the substrate metabolism for producing more reducing power was found to stimulate the production of several reduced thiol-containing proteins (including glutaredoxin and thioredoxin) that play key roles in Bio-QDs assembly. This effectively diverted the transformation route of the selenium (Se) and cadmium (Cd) metabolic from Cd3(PO4)2 formation to CdS xSe1- x QDs assembly, yielding fine-sized (2.0 ± 0.4 nm), high-quality Bio-QDs with quantum yield (5.2%) and fluorescence lifetime (99.19 ns) far exceeding the existing counterparts. The underlying mechanisms of Bio-QDs crystallization and development were elucidated by density functional theory calculations and molecular dynamics simulation. The resulting Bio-QDs were successfully used for bioimaging of cancer cells and tumor tissue of mice without extra modification. Our work provides fundamental knowledge on the Bio-QDs assembly mechanisms and proposes an effective, facile regulation strategy, which may inspire advances in controlled synthesis and practical applications of Bio-QDs as well as other bionanomaterials.
Assuntos
Cádmio/química , Imagem Molecular/métodos , Pontos Quânticos/química , Selênio/química , Animais , Cádmio/farmacologia , Sobrevivência Celular/efeitos dos fármacos , Escherichia coli/efeitos dos fármacos , Fluorescência , Glutarredoxinas/química , Glutarredoxinas/genética , Humanos , Camundongos , Microscopia de Fluorescência/métodos , Pontos Quânticos/metabolismo , Selênio/farmacologia , Especificidade por Substrato/efeitos dos fármacos , Tiorredoxinas/química , Tiorredoxinas/genéticaRESUMO
Ciprofloxacin (CIP), as an extensively used antibiotic, has been widely detected at a high level in the environment and has raised environmental pollution concerns. Thus, efficient and cost-effective methods for CIP degradation are highly desired. Biologically produced manganese oxides (BioMnOx) offer a promising perspective for CIP degradation because of their catalytic reactivity and cost-effectiveness. However, the release of Mn(II) from BioMnOx prevents the further oxidation of pollutants. As a consequence, continuous CIP degradation by BioMnOx is not feasible. In this work, a manganese redox cycling system driven by Pseudomonas putida MnB-1 was constructed for continuous degradation of CIP. In such a system CIP was oxidized continuously and rapidly by re-oxidizing the formed Mn(II) to regenerate reactive BioMnOx, which also protected the strain from CIP toxicity. CIP was degraded through N-dealkylation passway. No significant loss of BioMnOx reactivity was observed in three-cycle CIP degradation process, suggesting the stability of this system. An overlooked intracellular BioMnOx, which was involved in CIP degradation, was discovered in P. putida MnB-1. Moreover, the important role of Mn(III) in facilitating CIP removal in this system was also identified. This work provides useful information to better understand the degradation of antibiotic compounds mediated by microbes in environments.
Assuntos
Antibacterianos/metabolismo , Ciprofloxacina/metabolismo , Manganês/química , Manganês/metabolismo , Pseudomonas putida/metabolismo , OxirreduçãoRESUMO
Organisms served as factories of bio-assembly of nanoparticles attracted a lot of attentions due to the safe, economic and environmental-benignity traits, especially the fabrication of the super fluorescence properties quantum dots (QDs). However, information about the developmental dynamics of QDs in living organisms is still lacking. In this work, we synthesized cadmium-selenium (CdSe) QDs in Candida utilis WSH02-08, and then tracked and quantitatively characterized the developmental dynamics (photoactivation, photostable and photobleaching processes) of bio-QDs by translating fluorescence microscopy movies into visual quantitative curve. These findings shed light on the fluorescence properties of the bio-assembled QDs and are expected to accelerate the applications of the synthesized QDs in vivo. It provided a new way to screen bio-QDs and monitor the quality of QDs in vivo.
Assuntos
Compostos de Cádmio/metabolismo , Candida/metabolismo , Pontos Quânticos , Compostos de Selênio/metabolismo , Compostos de Cádmio/química , Fluorescência , Microscopia de Fluorescência , Pontos Quânticos/química , Pontos Quânticos/ultraestrutura , Compostos de Selênio/química , Espectrometria de FluorescênciaRESUMO
Nano-selenium has a great potential to be used in chemical, biological, medical and environmental fields. Biological methods for nano-selenium synthesis have attracted wide interests, because they can be operated at ambient temperature and pressure without complicated equipments. In this work, a protozoa, Tetrahymena thermophila (T. thermophila) SB210, was used to in vivo synthesize nano-selenium. The biosynthesized nano-selenium was characterized using transmission electron microscopy, energy dispersive X-ray spectroscopy and Raman spectroscopy. The synthesized amorphous spherical selenium nanoparticles had diameters of 50-500nm with the coexistence of irregular nano-selenium. The expressions of glutathione (GSH) synthesis related gene glutathione synthase, cysteine-rich protein metallothionein related gene metallothionein-1 and [2Fe-2S] cluster-binding protein related gene were up-regulated in the nano-selenium producing group. Also, the subsequent GSH detection and in vitro synthesis experimental results suggest the three proteins were likely to be involved in the nano-selenium synthesis process.
Assuntos
Nanopartículas Metálicas/química , Selênio/química , Tetrahymena thermophila/metabolismo , Biotecnologia , Genes de Protozoários , Glutationa/metabolismo , Glutationa Sintase/genética , Glutationa Sintase/metabolismo , Química Verde , Nanopartículas Metálicas/ultraestrutura , Nanotecnologia , Oxirredução , Proteínas de Protozoários/genética , Proteínas de Protozoários/metabolismo , Ácido Selenioso/metabolismo , Tetrahymena thermophila/genéticaRESUMO
Photothermal therapy (PTT) is a minimally invasive and effective cancer treatment method and has a great potential for innovating the conventional chemotherapy approaches. Copper sulfide (CuS) exhibits photostability, low cost, and high absorption in near infrared region, and is recognized as an ideal candidate for PTT. However, CuS, as a photothermal agent, is usually synthesized with traditional chemical approaches, which require high temperature, additional stabilization and hydrophilic modification. Herein, we report, for the first time, the preparation of CuS nanoparticles as a photothermal agent by a dissimilatory metal reducing bacterium Shewanella. oneidensis MR-1. The prepared nanoparticles are homogenously shaped, hydrophilic, small-sized (â¼5nm) and highly stable. Furthermore, the biosynthesized CuS nanoparticles display a high photothermal conversion efficiency of 27.2% because of their strong absorption at 1100nm. The CuS nanoparticles could be effectively used as a PTT agent under the irradiation of 1064nm. This work provides a simple, eco-friendly and cost-effective approach for fabricating PTT agents.