Engineering biomaterials for the recovery of rare earth elements.

The escalating global demand for rare earth elements (REEs) and the overabundance of REE-containing waste require innovative technologies for REE recovery from waste to achieve a sustainable supply of REEs while reducing the environmental burden. Biosorption mediated by peptides or proteins has emerged as a promising approach for selective REE recovery. To date, multiple peptides and proteins with high REE-binding affinity and selectivity have been discovered, and various strategies are being exploited to engineer robust and reusable biosorptive materials for selective REE recovery. This review highlights recent advances in discovering and engineering peptides and proteins for REE recovery. Future research prospects and challenges are also discussed.

Lanmodulin-Functionalized Magnetic Nanoparticles as a Highly Selective Biosorbent for Recovery of Rare Earth Elements.

Recovering rare earth elements (REEs) from waste streams represents a sustainable approach to diversify REE supply while alleviating the environmental burden. However, it remains a critical challenge to selectively separate and concentrate REEs from low-grade waste streams. In this study, we developed a new type of biosorbent by immobilizing Lanmodulin-SpyCatcher (LanM-Spycatcher) on the surface of SpyTag-functionalized magnetic nanoparticles (MNPs) for selective separation and recovery of REEs from waste streams. The biosorbent, referred to as MNP-LanM, had an adsorption activity of 6.01 ± 0.11 µmol-terbium/g-sorbent and fast adsorption kinetics. The adsorbed REEs could be desorbed with >90% efficiency. The MNP-LanM selectively adsorbed REEs in the presence of a broad range of non-REEs. The protein storage stability of the MNP-LanM increased by two-fold compared to free LanM-SpyCatcher. The MNP-LanM could be efficiently separated using a magnet and reused with high stability as it retained ∼95% of the initial activity after eight adsorption-desorption cycles. Furthermore, the MNP-LanM selectively adsorbed and concentrated REEs from the leachate of coal fly ash and geothermal brine, resulting in 967-fold increase of REE purity. This study provides a scientific basis for developing innovative biosorptive materials for selective and efficient separation and recovery of REEs from low-grade feedstocks.

Distribution of the new functional marker gene (pahE) of aerobic polycyclic aromatic hydrocarbon (PAHs) degrading bacteria in different ecosystems.

Understanding the degradation potentials in PAHs-contaminated sites is significant for formulating effective bioremediation strategies. pahE encoding PAHs hydratase-aldolase has been proven as a better new functional marker gene of aerobic PAHs-degrading bacteria to assess the biodegradation potential of indigenous PAHs-degrading bacterial population. However, the distribution of pahE and its relationship with environmental factors remain unknown. The present study observed spatial variations in the diversity and abundance of pahE across oilfield soils, mangrove sediments, and urban roadside soils. nahE from Pseudomonas, bphE from Hyphomonas oceanitis, nagE from Comamonas testosterone, and novel pahE genes were widely present in these PAHs-polluted ecosystems. The abundance of pahE in PAHs-contaminated sites was in the range of 105-106 copies·g-1 (dry weight). Redundancy analysis and Pearson's correlation analysis implied that the distribution of pahE in the PAHs-contaminated environment was mainly shaped by environmental factors such as PAHs pollution level, nutrient level, salinity, and water content. This work was the first to explore the distribution of the new functional marker gene (pahE) and its links with environmental parameters, which provided new insights into the ecophysiology and distribution of indigenous aerobic PAHs-degrading bacteria in contaminated sites.

Site-Specific and Tunable Co-immobilization of Proteins onto Magnetic Nanoparticles via Spy Chemistry.

Co-immobilization of multiple proteins onto one nanosupport has large potential in mimicking natural multiprotein complexes and constructing efficient cascade biocatalytic systems. However, control of different proteins regarding their spatial arrangement and loading ratio remains a big challenge, and protein co-immobilization often requires the use of purified proteins. Herein, built upon our recently designed SpyTag-functionalized magnetic nanoparticles (MNPs), we established a modular MNP platform for site-specific, tunable, and cost-effective protein co-immobilization. SpyCatcher-fused enhanced green fluorescent protein (i.e., EGFP-SpyCatcher) and mCherry red fluorescent protein (i.e., RFP-SpyCatcher) were designed and conjugated on MNPs, and the immobilized proteins showed 3-7-fold enhancement in storage stability and greatly improved stability against the freeze-thaw process compared to free proteins. The protein-conjugated MNPs also retained desirable colloidal stability and magnetic responsiveness, enabling facile proteins' recovery. Also, one-pot co-immobilization of the two proteins could be fine-tuned with their feed ratios. In addition, MNPs could selectively and efficiently co-immobilize both SpyCatcher-fused proteins from combined cell lysates without purification, offering a convenient and cost-effective approach for multiprotein immobilization. This MNP platform provides a facile and efficient tool to construct bionano hybrid materials (i.e., protein-based MNPs) and multiprotein systems for a variety of industrial and green chemistry applications.

Lake reclamation alters molecular-level characteristics of lacustrine dissolved organic matter - A study of nine lakes in the Yangtze Plain, China.

In recent decades, the reclamation of lakes has captured 42% of the total lake area of the Yangtze Plain in China and introduced additional pressure on lacustrine water quality. While lacustrine dissolved organic matter (DOM) is critical in regulating biogeochemical processing and aquatic biodiversity, the impact of reclamation on the molecular-level characteristics of lacustrine DOM remains unexplored. Here, the DOM characteristics altered by reclamation practices in the Yangtze Plain lakes were investigated using fluorescence spectroscopy, nuclear magnetic resonance spectroscopy, and Fourier transform ion cyclotron resonance mass spectrometry. Results demonstrated that reclamation not only elevated the quantity (on average +32%) but also altered the characteristics and composition of lacustrine DOM. Compared to the natural water sites close by, reclamation sites did not significantly alter the DOM aromaticity but significantly lowered the average molecular weight and increased the biolability of DOM. The chromophoric DOM and humic-like fluorescent components were remarkably elevated, but not the protein-like fluorescent components. More lipid-like and condensed aromatic-like components were detected in the lacustrine DOM as compared to the lignin-like, carbohydrate-like, and protein-like components, which may be driven by the increased microbial processing. Overall, the significant alteration in characteristics and composition of lacustrine DOM highlights the potential impact of reclamation on the DOM biogeochemical cycle and the environmental quality in aquatic ecosystems.

Shifts of the new functional marker gene (pahE) of polycyclic aromatic hydrocarbons (PAHs) degrading bacterial population and its relationship with PAHs biodegradation.

Identification of polycyclic aromatic hydrocarbons (PAHs) degrading bacterial populations and understanding their responses to PAHs are crucial for the designing of appropriate bioremediation strategies. In this study, the responses of PAHs-degrading bacterial populations to different PAHs were studied in terms of the compositions and abundance variations of their new functional marker gene (pahE) by gene-targeted metagenomic and qPCR analysis. Overall, PAHs species significantly affected the composition and abundance of pahE gene within the PAHs-degrading bacteria in each treatment and different pahE of PAHs-degrading bacteria involved in the different stages of PAHs degradation. Noted that new pahE genotypes were also discovered in all PAHs treatment groups, indicating that some potential new PAHs-degrading bacterial genera were also involved in PAHs degradation. Besides, all three PAH removal rates were significantly positively related with pahE gene abundances (R2 = 0.908 ~ 0.922, p < 0.01), demonstrating that pahE could be a good indicator of PAHs degradation activity or potential. This is the first study focusing on the dynamic changes of the pahE gene within PAHs-degrading bacterial community during the degradation of PAHs in mangrove sediment, providing novel insights into the use of pahE gene as the functional marker to indicate PAH degradation.

Magnetic Nanoplatforms for Covalent Protein Immobilization Based on Spy Chemistry.

Immobilization of proteins on magnetic nanoparticles (MNPs) is an effective approach to improve protein stability and facilitate separation of immobilized proteins for repeated use. Herein, we exploited the efficient SpyTag-SpyCatcher chemistry for conjugation of functional proteins onto MNPs and established a robust magnetic-responsive nanoparticle platform for protein immobilization. To maximize the loading capacity and achieve outstanding water dispersity, the SpyTag peptide was incorporated into the surface-charged polymers of MNPs, which provided abundant active sites for Spy chemistry while maintaining excellent colloidal stability in buffer solution. Conjugation between enhanced green fluorescence protein (EGFP)-SpyCatcher-fused proteins and SpyTag-functionalized MNPs was efficient at ambient conditions without adding enzymes or chemical cross-linkers. Benefiting from the excellent water dispersity and interface compatibility, the surface Spy reaction has fast kinetics, which is comparable to that of the solution Spy reaction. No activity loss was observed on EGFP after conjugation due to the site-selective nature of Spy chemistry. The immobilization process of EGFP on MNPs was highly specific and robust, which was not affected by the presence of other proteins and detergents, such as bovine serum albumin and Tween 20. The MNP platform was demonstrated to be protective to the conjugated EGFP and significantly improved the shelf life of immobilized proteins. In addition, experiments confirmed the retained magnetophoresis of the MNP after protein loading, demonstrating fast MNP recovery under an external magnetic field. This MNP is expected to provide a versatile and modular platform to achieve effective and specific immobilization of other functional proteins, enabling easy reuse and storage.

Spectroscopic and molecular-level characteristics of dissolved organic matter in the Pearl River Estuary, South China.

Coastal populations are expanding globally, resulting in great anthropogenic impacts on the organic matter in estuaries and regional carbon cycles. However, the molecular-level characteristics of dissolved organic matter (DOM) within highly disturbed estuaries are still not well understood. Here, water samples collected during two seasons (wet and dry) from the subtropical Pearl River Estuary of China were analyzed using absorption and fluorescence spectroscopy and Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR MS) to determine the spatiotemporal variations in the DOM characteristics. In the seaward direction, the abundances of chromophoric and fluorescent DOM decreased by greater percentages than the bulk dissolved organic carbon concentration. The spectroscopy and FT-ICR MS analyses collectively indicated seaward declines in the aromaticity of DOM and terrestrial DOM contributions from natural terrestrial markers and anthropogenic synthetic surfactants. In particular, the S content in DOM was much higher here than in previously reported estuaries, suggesting a strong anthropogenic impact on the estuarine DOM. Greater terrestrial and anthropogenic signatures in DOM were observed in the wet season than in the dry season. Importantly, this study implies that the terrestrial and anthropogenic contributions to DOM were strongly driven by season in the anthropogenically disturbed subtropical estuary.

Molecular characterization of methanogenic microbial communities for degrading various types of polycyclic aromatic hydrocarbon.

Knowledge on methanogenic microbial communities associated with the degradation of polycyclic aromatic hydrocarbons (PAHs) is crucial to developing strategies for PAHs bioremediation. In this study, the linkage between the type of PAHs and microbial community structure was fully investigated through 16S rRNA gene sequencing on four PAH-degrading cultures. Putative degradation products were also detected. Our results indicated that naphthalene (Nap)/2-methylnaphthalene (2-Nap), phenanthrene (Phe) and anthracene (Ant) sculpted different microbial communities. Among them, Nap and 2-Nap selected for similar degrading bacteria (i.e., Alicycliphilus and Thauera) and methanogens (Methanomethylovorans and Methanobacterium). Nap and 2-Nap were probably activated via carboxylation, producing 2-naphthoic acid. In contrast, Phe and Ant shaped different bacterial and archaeal communities, with Arcobacter and Acinetobacter being Phe-degraders and Thiobacillus Ant-degrader. Methanogenic archaea Methanobacterium and Methanomethylovorans predominated Phe-degrading and Ant-degrading culture, respectively. These findings can improve our understanding of natural PAHs attenuation and provide some guidance for PAHs bioremediation in methanogenic environment.

Assessment of extracellular antibiotic resistance genes (eARGs) in typical environmental samples and the transforming ability of eARG.

The emergence and spread of antibiotic resistance has pose a huge threat to both human health and environmental ecosystem. However, little is known regarding the pool of ARGs in extracellular DNA (eDNA). In this study ten ARGs (sul1, sul2, tetW, tetX, ermA, ermB, blaTEM, ampC, cat and cmr) and class I integron (intI1) in the sludge from hospital, pharmaceutical industry, wastewater treatment plant (WWTP), and swine manure, and sediment in urban lake in the form of both eDNA and intracellular DNA (iDNA) were evaluated by quantitative polymerase chain reaction (qPCR). The results showed that every gram of sludge dry weight contained from 7.31 × 103 to 1.16 × 1010 copies of extracellular ARGs (eARGs) and from 1.04 × 105 to 2.74 × 1012 copies of intracellular ARGs (iARGs). The sludge from hospital with the highest ratio of eARGs to total ARGs (11.02-89.63%), followed by the sediment from urban lake, implying that most of the ARGs in these regions were contributed by eARGs. The relative abundance of eARGs were higher than iARGs in sludge from WWTP and pharmaceutical industry, moreover, 1/3 and 5/9 detected eARGs were higher than the ARGs in the iDNA extracted from sludge of hospital and sediment from urban lake, respectively. Furthermore, the transforming ability of eARGs suggesting that adsorbed eARG is more preferentially coupled to the competent cells than free eARG. These findings highlight the need to focus attention on the contribution of eARGs to the dissemination of antibiotic resistance into environment, and also future needs in mitigating the spread of eARGs in the environment.

Distribution of bacterial polycyclic aromatic hydrocarbon (PAH) ring-hydroxylating dioxygenases genes in oilfield soils and mangrove sediments explored by gene-targeted metagenomics.

PAH ring-hydroxylating dioxygenases (PAH-RHDα) gene, a useful biomarker for PAH-degrading bacteria, has been widely used to examine PAH-degrading bacterial community in different contaminated sites. However, the distribution of PAH-RHDα genes in oilfield soils and mangrove sediments and their relationship with environmental factors still remain largely unclear. In this study, gene-targeted metagenomics was first used to investigate the diversity of PAH-degrading bacterial communities in oilfield soils and mangrove sediments. The results showed that higher diversity of PAH-degrading bacteria in the studied samples was revealed by gene-targeted metagenomics than traditional clone library analysis. Pseudomonas, Burkholderia, Ralstonia, Polymorphum gilvum, Mycobacterium, Sciscionella marina, Rhodococcus, and potential new degraders were prevailed in the oilfield area. For mangrove sediments, novel PAH degraders and Mycobacterium were predominated. The spatial distribution of PAH-RHDα gene was dependent on geographical location and regulated by local environmental variables. PAH content played a key role in shaping PAH-degrading bacterial communities in the studied samples, which would enrich PAH-degrading bacterial population and decrease PAH-degrading bacterial diversity. This work brings a more comprehensive and some new insights into the distribution and biodegradation potential of PAH-degrading bacteria in soil and sediments ecosystems.

Distribution comparison and risk assessment of free-floating and particle-attached bacterial pathogens in urban recreational water: Implications for water quality management.

The risk of pathogen exposure in recreational water is a concern worldwide. Moreover, suspended particles, as ideal shelters for pathogens, in these waters also need attention. However, the risk caused by the pathogen-particle attachment is largely unknown. Accordingly, water samples in three recreational lakes in Beijing were collected and separated into free-floating (FL, 0.22-5µm) and particle-attached (PA, >5µm) fractions. Next-generation sequencing (NGS) was employed to determine the diversity of genera containing pathogens, and quantitative PCR (qPCR) was used to assess the presence of genes from Escherichia coli (uidA), Salmonella enterica (invA), Aeromonas spp. (aerA), Mycobacterium avium (16S) and Pseudomonas aeruginosa (oaa). The NGS results showed stable pathogen genera composition distinctions between the PA and FL fractions. Some genera, such as Aeromonas and Mycobacterium, exhibited higher abundances in the PA fractions. qPCR revealed that most of the gene concentrations were higher within particles than were FL fractions. Some gene levels showed correlations with the particle concentrations and lake nutrient levels. Further quantitative microbial risk assessment (QMRA) of selected strains (S. enterica and M. avium) indicated a higher health risk during secondary contact activities in lakes with more nutrients and particles. We concluded that suspended particles (mainly composed of algae) in urban recreational water might influence the pathogen distribution and could serve as reservoirs for pathogen contamination, with important management implications.