Generation of a Gluconobacter oxydans knockout collection for improved extraction of rare earth elements.
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ABSTRACT:
SUBMITTER: Schmitz AM
PROVIDER: S-EPMC8602642 | biostudies-literature |
REPOSITORIES: biostudies-literature
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Project description:In Gluconobacter oxydans, pyrroloquinoline quinone (PQQ) serves as the cofactor for various membrane-bound dehydrogenases that oxidize sugars and alcohols in the periplasm. Proteins for the biosynthesis of PQQ are encoded by the pqqABCDE gene cluster. Our reverse transcription-PCR and promoter analysis data indicated that the pqqA promoter represents the only promoter within the pqqABCDE cluster of G. oxydans 621H. PQQ overproduction in G. oxydans was achieved by transformation with the plasmid-carried pqqA gene or the complete pqqABCDE cluster. A G. oxydans mutant unable to produce PQQ was obtained by site-directed disruption of the pqqA gene. In contrast to the wild-type strain, the pqqA mutant did not grow with d-mannitol, d-glucose, or glycerol as the sole energy source, showing that in G. oxydans 621H, PQQ is essential for growth with these substrates. Growth of the pqqA mutant, however, was found with d-gluconate as the energy source. The growth behavior of the pqqA mutant correlated with the presence or absence of the respective PQQ-dependent membrane-bound dehydrogenase activities, demonstrating the vital role of these enzymes in G. oxydans metabolism. A different PQQ-deficient mutant was generated by Tn5 transposon mutagenesis. This mutant showed a defect in a gene with high homology to the Escherichia coli tldD gene, which encodes a peptidase. Our results indicate that the tldD gene in G. oxydans 621H is involved in PQQ biosynthesis, possibly with a similar function to that of the pqqF genes found in other PQQ-synthesizing bacteria.
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Project description:Rare earth elements (REEs) are critical materials in electronics and clean technologies. With the diminishing of easily accessible minerals for mining, the REE recovery from waste is an alternative toward a circular economy. Present methods for REE recovery suffer from lengthy purifications, low extractability, and high wastewater streams. Here, we report an ultrafast electrothermal process (~3000°C, ~1 s) based on flash Joule heating (FJH) for activating wastes to improve REE extractability. FJH thermally degrades or reduces the hard-to-dissolve REE species to components with high thermodynamic solubility, leading to ~2× increase in leachability and high recovery yields using diluted acid (e.g., 0.1 M HCl). The activation strategy is feasible for various wastes including coal fly ash, bauxite residue, and electronic waste. The rapid FJH process is energy-efficient with a low electrical energy consumption of 600 kWh ton-1. The potential for this route to be rapidly scaled is outlined.
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Project description:The acetic acid bacterium Gluconobacter oxydans 621H is characterized by its exceptional ability to incompletely oxidize a great variety of carbohydrates in the periplasm. The metabolism of this ?-proteobacterium has been characterized to some extent, yet little is known about its transcriptomes and related data. In this study, we applied two different RNAseq approaches. Primary transcriptomes enriched for 5'-ends of transcripts were sequenced to detect transcription start sites, which allow subsequent analysis of promoter motifs, ribosome binding sites, and 5´-UTRs. Whole transcriptomes were sequenced to identify expressed genes and operon structures.Sequencing of primary transcriptomes of G. oxydans revealed 2449 TSSs, which were classified according to their genomic context followed by identification of promoter and ribosome binding site motifs, analysis of 5´-UTRs including validation of predicted cis-regulatory elements and correction of start codons. 1144 (41%) of all genes were found to be expressed monocistronically, whereas 1634 genes were organized in 571 operons. Together, TSSs and whole transcriptome data were also used to identify novel intergenic (18), intragenic (328), and antisense transcripts (313).This study provides deep insights into the transcriptional landscapes of G. oxydans. The comprehensive transcriptome data, which we made publicly available, facilitate further analysis of promoters and other regulatory elements. This will support future approaches for rational strain development and targeted gene expression in G. oxydans. The corrections of start codons further improve the high quality genome reference and support future proteome analysis.
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Project description:Due to the increasing demand of Rare Earth Elements (REE or RE), new and more efficient techniques for their extraction are necessary, suitable for both mining and recycling processes. Current techniques such as solvent extraction or solid adsorbents entail drawbacks such as using big volumes of harmful solvents or limited capacity. Hybrid nanoadsorbents based on SiO2 and highly stable γ-Fe2O3-SiO2 nanoparticles, proved recently to be very attractive for adsorption of REE, yet not being the absolute key to solve the problem. In the present work, we introduce a highly appealing new approach in which the nanoparticles, rather than behaving as adsorbent materials, perform as inducers of crystallization for the REE in the form of hydroxides, allowing their facile and practically total removal from solution. This induced crystallization is achieved by tuning the pH, offering an uptake efficiency more than 20 times higher than previously reported (up to 900 mg RE3+/g vs. 40 mg RE3+/g). The obtained phases were characterized by SEM-EDS, TEM, STEM and EFTEM and 13C and 29Si solid state NMR. Magnetic studies showed that the materials possessed enough magnetic properties to be easily removed by a magnet, opening ways for an efficient and industrially applicable separation technique.
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Project description:Rare-earth elements (REEs) are involved in most high technology devices and have become critical for many countries. The progress of processes for the extraction and recovery of REEs is therefore essential. Liquid-solid extraction methods are an attractive alternative to the conventional solvent extraction process used for the separation and/or purification of REEs. For this purpose, a solid-phase extraction system was investigated for the extraction and valorization of REEs. Ion-exchange resins were synthesized involving the condensation of terephthalaldehyde with resorcinol under alkaline conditions. The terephthalaldehyde, which is a non-hazardous aromatic dialdehyde, was used as an alternative to formaldehyde that is toxic and traditionally involved to prepare phenolic ion-exchange resins. The resulting formaldehyde-free resole-type phenolic resins were characterized and their ion-exchange capacity was investigated in regard to the extraction of rare-earth elements. We herein present a promising formaldehyde and phenol-free as a potential candidate for solid-liquid extraction REE with a capacity higher than 50 mg/g and the possibility to back-extract the REEs by a striping step using a 2 M HNO3 solution.
| S-EPMC8780303 | biostudies-literature
Project description:The task-specific ionic liquid trihexyltetradecylphosphonium 3-hydroxy-2-naphthoate has been described as a suitable extraction agent for numerous metals from aqueous phases, while additionally providing reduced leaching into the used matrices. Here, we investigate the extraction properties of this extractant towards rare earth elements. Of these, La, Ce, Nd, Ho und Lu were chosen as a representative mix of light and heavy elements. Single- as well as double-element extractions were carried out under varying conditions regarding pH, temperature and extraction time. The highest extraction efficacies and minimalized precipitation of the respective metals were recorded at a pH of 2.5. Satisfactory extraction efficacies (>80%) were achieved already after 6 hours for the elements Ce, Nd and Lu in single-element extraction experiments at room temperature. Increased temperatures improved the extraction efficacy for Nd from 36% at 20 °C to 80% at 30 °C after only 2 hours. Surprisingly, this effect was not observed for Ce in single-element experiments. In double-element feed solutions containing both Ce and Nd, however, the time-dependant extraction efficacy of Ce mirrored that of Nd. The pH in the aqueous extraction matrix changed during the extraction, showing a positive correlation with the extraction efficacy and revealing the extraction mechanism to be via anion exchange. The leaching was in good agreement with literature values, showed a positive correlation with extraction efficacies, and ranged for all extractions between 0.8 and 1.2%. Remarkably, increasing the temperature from 20 °C to 30 °C had no significant influence on leaching.
| S-EPMC10440728 | biostudies-literature
Project description:With the growing need for high-purity rare-earth elements (REEs), the separation of these REEs has received much attention recently. The objective of this research is to produce chitosan from shrimp waste, then modify it with different functionality, and investigate the adsorption properties of chitosan adsorbents towards La(III) ions. First, from shrimp waste, chitosan (ch) with a significant degree of deacetylation, purity, and solubility was produced. The purified chitosan was cross-linked with epichlorohydrin (ep), and then, it was modified with 3,6,9,12-tetraazatetradecane-1,14-diamine (HA) to produce polyaminated chitosan (HA@ep@Ch). The polycarboxylated/imine chitosan (CM@HA@ep@Ch) was obtained by treating polyaminated chitosan with chloroacetic acid in isopropyl alcohol. The chitosan adsorbents were characterized and applied for lanthanum recovery from synthetic and monazite leach liquor samples. The factors controlling the recovery process were studied and discussed. The performance of the adsorbents was achieved through equilibrium, dynamic, and isothermal studies. HA@ep@Ch and CM@HA@ep@Ch showed good performance for lanthanum recovery with a maximum capacity of 114.52 and 141.76 mg/g at 330 K, respectively. The isotherm parameters refer to the monolayer of lanthanum adsorbed into the adsorbents through chelation and ion exchange mechanisms. A 0.5-M HCl solution was found effective to elute 95.8% of the adsorbed lanthanum on HA@ep@Ch, and 93.4% of the adsorbed lanthanum on CM@HA@ep@Ch. The adsorbents showed greater selectivity in extracting La, Ce, Pr, Nd, and Sm (62-75%) from REE leach liquid compared to extracting other REEs (20-41%).
| S-EPMC10611849 | biostudies-literature
Project description:Until recently, rare-earth elements (REEs) had been thought to be biologically inactive. This view changed with the discovery of the methanol dehydrogenase (Mdh) XoxF that strictly relies on REEs for its activity. Some methylotrophs only contain xoxF, while others, including the model phyllosphere colonizer Methylobacterium extorquens PA1, harbor this gene in addition to mxaFI encoding a Ca2+-dependent enzyme. Here we found that REEs induce the expression of xoxF in M. extorquens PA1, while repressing mxaFI, suggesting that XoxF is the preferred Mdh. Using reporter assays and a suppressor screen, we found that La3+ is sensed both in a XoxF-dependent and independent manner. Furthermore, we investigated the role of REEs during Arabidopsis thaliana colonization. Element analysis of the phyllosphere revealed the presence of several REEs at concentrations up to 10 μg per g dry weight. Complementary proteome analyses of M. extorquens PA1 revealed XoxF as a top induced protein in planta and resulted in the identification of a core set of La3+-regulated proteins under defined artificial media conditions. Among these, we identified a potential REE-binding protein that is encoded next to a gene for a TonB-dependent transporter. The latter was essential for REE-dependent growth on methanol indicating chelator-assisted uptake of REEs.
2019-01-21 | PXD011842 | Pride
Project description:Gluconobacter oxydans Raw sequence reads
| PRJNA299470 | ENA