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Microbial metabolism is an important driving force for the elimination of 4-chlorophenoxyacetic acid residues in the environment. The α-Ketoglutarate-dependent dioxygenase (TfdA) or 2,4-D oxygenase (CadAB) catalyzes the cleavage of the aryl ether bond of 4-chlorophenoxyacetic acid to 4-chlorophenol, which is one of the important pathways for the initial metabolism of 4-chlorophenoxyacetic acid by microorganisms. However, strain Cupriavidus sp. DL-D2 could utilize 4-chlorophenoxyacetic acid but not 4-chlorophenol for growth. This scarcely studied degradation pathway may involve novel enzymes that has not yet been characterized. Here, a gene cluster (designated cpd) responsible for the catabolism of 4-chlorophenoxyacetic acid in strain DL-D2 was cloned and identified, and the dioxygenase CpdA/CpdB responsible for the initial degradation of 4-chlorophenoxyacetic acid was successfully expressed, which could catalyze the conversion of 4-chlorphenoxyacetic acid to 4-chlorocatechol. Then, an aromatic cleavage enzyme CpdC further converts 4-chlorocatechol into 3-chloromuconate. The results of substrate degradation experiments showed that CpdA/CpdB could also degrade 3-chlorophenoxyacetic acid and phenoxyacetic acid, and homologous cpd gene clusters were widely discovered in microbial genomes. Our findings revealed a novel degradation mechanism of 4-chlorophenoxyacetic acid at the molecular level., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

The utility of endophytic bacteria in Cadmium (Cd) remediation has gained significant attention due to their ability to alleviate metal-induced stress and enhance plant growth. Here, we investigate C. metallidurans CML2, an endophytic bacterial strain prevalent in rice, showing resilience against 2400 mg/L of Cd(II). We conducted an in-depth integrated morphological and transcriptomic analysis illustrating the multifarious mechanisms CML2 employs to combat Cd, including the formation of biofilm and CdO nanoparticles, upregulation of genes involved in periplasmic immobilization, and the utilization of RND efflux pumps to extract excess Cd ions. Beyond Cd, CML2 exhibited robust tolerance to an array of heavy metals, including Mn 2+ , Se 4+ , Ni 2+ , Cu 2+ , and Hg 2+ , demonstrating effective Cd(II) removal capacity. Furthermore, CML2 has exhibited plant growth-promoting properties through the production of indole-3-acetic acid (IAA) at 0.93 mg/L, soluble phosphorus compounds at 1.11 mg/L, and siderophores at 22.67%. Supportively, pot experiments indicated an increase in root lengths and a decrease in Cd bioaccumulation in rice seedlings inoculated with CML2, consequently reducing Cd translocation rates from 43% to 31%. These findings not only contribute to the understanding of Cd resistance mechanisms in C. metallidurans, but also underscore CML2's promising application in Cd remediation within rice farming ecosystems., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

This study investigated an effective strategy for remediating antimony (Sb)-contaminated soil using the bacterial strain screened from Sb-contaminated fern rhizospheres due to its superior growth-promoting, heavy-metal(loid) resistant, and antibiotic-tolerant characteristics. The strain that belongs to Cupriavidus sp. was determined by 16S rRNA sequencing and showed no morphological changes when grown with high concentrations of Sb (608.8 mg/L). The strain showed prominent indole acetic acid (IAA), phosphate-solubilizing abilities, and ACC deaminase activity under Sb stress. Moreover, IAA and soluble phosphate levels increased in the presence of 608.8 mg/L Sb. Inoculation of rape seedlings with Cupriavidus sp. S-8-2 enhanced several morphological and biochemical growth features compared to untreated seedlings grown under Sb stress. Inoculation of Cupriavidus sp. S-8-2 increased root weight by more than four-fold for fresh weight and over two-fold for dry weight, despite high environmental Sb. The strain also reduced Sb-mediated oxidative stress and malondialdehyde contents by reducing Sb absorption, thus alleviating Sb-induced toxicity. Environmental Scanning Electron Microscope (ESEM) imaging and dilution plating technique revealed Cupriavidus sp. S-8-2 is localized on the surface of roots. Identifying the Sb-resistant plant growth-promoting bacterium suggested its usefulness in the remediation of contaminated agricultural soil and for the promotion of crop growth. We highly recommend the strain for further implementation in field experiments., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022. Published by Elsevier B.V.)

This study focused on a novel denitrifying Cupriavidus sp. W12, which can perform microbial induced calcium precipitation (MICP) to remove fluoride (F - ) under aerobic and anaerobic conditions. Under anaerobic condition, the removal ratios of F - , calcium (Ca 2+ ), and nitrate (NO 3 - -N) reached 87.52%, 65.03%, and 96.06%, respectively, which were higher than that under aerobic condition (50.17%, 88.21%, and 67.33%, respectively). Higher pH of 8.26 was obtained after 120 h of the strain W12 growth under anaerobic condition than that under aerobic condition (7.77). The F - removal ratio of 98.20% was predicted by the response surface methodology (RSM). Scanning electron microscopy (SEM) images of anaerobic precipitation were dense and porous. CaCO 3 , Ca 5 (PO 4 ) 3 OH, Ca 5 (PO 4 ) 3 F, and CaF 2 were determined by X-ray diffraction (XRD) and energy dispersive spectrometer (EDS). Self-aggregation of bacteria and adsorption of biological crystal seeds were the determinant of the precipitates formation. The results of infrared spectrometer (FTIR) and excitation-emission matrix (EEM) showed that anaerobic extracellular polymeric substances (EPS) expression led the proportion of hydroxylapatite in the precipitates increased. As the first report on the anaerobic MICP to remove F - , it provides a theoretical basis for the remediation of F - , Ca 2+ , and NO 3 - -N in groundwater., (Copyright © 2021 Elsevier B.V. All rights reserved.)

The chiral pesticide enantiomers often show selective efficacy and non-target toxicity. In this study, the enantioselective degradation characteristics of the chiral organophosphorus insecticide isocarbophos (ICP) by Cupriavidus nantongensis X1 T were investigated systematically. Strain X1 T preferentially degraded the ICP R isomer (R-ICP) over the S isomer (S-ICP). The degradation rate constant of R-ICP was 42-fold greater than S-ICP, while the former is less bioactive against pest insects but more toxic to humans than the latter. The concentration ratio of S-ICP to R-ICP determines whether S-ICP can be degraded by strain X1 T . S-ICP started to degrade only when the ratio (C S-ICP /C R-ICP ) was greater than 62. Divalent metal cations could improve the degradation ability of strain X1 T . The detected metabolites that were identified suggested a novel hydrolysis pathway, while the hydrolytic metabolites were less toxic to fish and green algae than those from P-O bond breakage. The crude enzyme degraded both R-ICP and S-ICP in a similar rate, indicating that enantioselective degradation was due to the transportation of strain X1 T . The strain X1 T also enantioselectively degraded the chiral organophosphorus insecticides isofenphos-methyl and profenofos. The enantioselective degradation characteristics of strain X1 T make it suitable for remediation of chiral organophosphorus insecticide contaminated soil and water., (Copyright © 2021 Elsevier B.V. All rights reserved.)

The co-contamination of groundwater with nitrate (NO 3 -N) and manganese (Mn(II)) is a global issue that needs to be efficiently remediated. In this research, a novel denitrifying and manganese-oxidizing strain HY129 was isolated from the sediments sample of a drinking water and identified as Cupriavidus sp. HY129. The remediation ability of strain HY129 regarding the nitrate and Mn(II) pollution were investigated. The removal efficiency of nitrate and Mn(II) were 99.81% (0.229 mgL - -N) and manganese (Mn(II)) is a global issue that needs to be efficiently remediated. In this research, a novel denitrifying and manganese-oxidizing strain HY129 was isolated from the sediments sample of a drinking water and identified as Cupriavidus sp. HY129. The remediation ability of strain HY129 regarding the nitrate and Mn(II) pollution were investigated. The removal efficiency of nitrate and Mn(II) were 99.81% (0.229 mgL -1 h -1 ) and 87.24% (0.233 mgL -1 h -1 ) in bacterial culture after 72 h, respectively. Moreover, the addition of Mn(II) significantly enhanced the denitrification process, while excessive concentration of Mn(II) caused more NO 2 - -N accumulation. The impacts of adsorption and oxidation activity on Mn(II) removal were investigated. Protein in extracellular polymeric substance (EPS) which produced in the Mn-oxidizing process was speculated to be the main cause of extracellular adsorption of Mn(II). Characterization of biogenic manganese oxides (BMO) confirmed the formation of high-valent manganese and the trapping experiment with sodium pyrophosphate (NaPP) demonstrated the existence of Mn(III)-intermediates. Furthermore, multicopper oxidase gene amplification provided evidence for the molecular biology of Mn(II) oxidation by strain HY129., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Chlorpyrifos is an important organophosphorus insecticide. It is highly toxic to mammals and can pollute the environment. Cupriavidus nantongensis X1 T can efficiently degrade chlorpyrifos. Immobilization technology can also improve the viability, stability and catalytic ability of bacteria. In this study, strain X1 T was, therefore, captured on various composite immobilized carriers, sodium alginate (SA), diatomite (KLG), chitosan (CTS) and polyvinyl alcohol (PVA). The four types of immobilized beads (SA, SA + KLG, SA + CTS and SA + PVA) could form a slice and honeycomb structure to capture strain X1 T . The results showed that SA + CTS (SC) was an optimal material combination for the immobilization of strain X1 T to degrade chlorpyrifos. Compared with SA-X1 T , after adding CTS, the specific surface area and adsorption capacity for chlorpyrifos were increased 3.4 and 1.7 fold, respectively. SC-X1 T could degrade 96.6% of chlorpyrifos at 20 mg/L within 24 h and the degradation rate constant was 4.8 fold greater than immobilized strain LLBD2, a well-studied chlorpyrifos-degrading strain. The immobilized beads SC-X1 T also showed a more stable and greater degradation ability than X1 T free cells for chlorpyrifos in industrial wastewater. The synergy of adsorption and degradation of immobilized strain X1 T is suitable for in-situ remediation of chlorpyrifos contaminated environment., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Heavy metal contamination is becoming a global problem threatening human health. Heavy metal removal by engineered microbes by cellular adsorption and uptake is a promising strategy for treatment of heavy metal contamination. However, this strategy is confronted with limited heavy metal-capturing elements. In this study, we performed a transcription profiling-guided strategy for construction of heavy metal-capturing synthetic bacteria. Transcription profiling of a heavy metal-tolerating Cupriavidus taiwanensis strain revealed up-regulation of sulfur metabolism-related operons (e.g., iscSAU and moaEDAB) by Pb 2+ and Cd 2+ . A synthetic Escherichia coli strain, EcSSMO, was constructed by design of a synthetic sulfur metabolism operon (SSMO) based on iscSAU/moaEDAB. Biochemical analysis and X-ray photoelectron spectroscopy (XPS) revealed that the synthetic bacteria had remodeled sulfur metabolism and enhanced heavy metal-tolerating capacity, with higher surviving EcSSMO cells than the surviving control cells Ec0 (not containing SSMO) at 50 mg/L of Pb 2+ and Cd 2+ (>92 % versus <10 %). Moreover, EcSSMO exhibited much higher heavy metal-capturing capacity than Ec0, removing>90 % of Pb 2+ and Cd 2+ at 5 mg/L of Pb 2+ and Cd 2+ , and >40 % of both heavy metals even at 50 mg/L of Pb 2+ and Cd 2+ . This study reveals emphasizes feasibility of transcription profiling-guided construction of synthetic organisms by large-scale remodeling metabolic network., (Copyright © 2020. Published by Elsevier B.V.)

Nitrobenzene, nitrotoluenes and nitrobenzoic acid are toxic and mutagenic. Their removal from the environment is necessary to avoid health and environmental damage. In this study, Cupriavidus strain a3 was found to utilize 2-nitrotoluene (2NT), 3-nitrotoluene (3NT), 4-nitrotoluene (4NT), nitrobenzene (NB) and 2-nitrobenzoic acid (2NBA) as carbon and nitrogen source, resulting in their detoxification. The metabolism involved reductive transformation of nitroaromatics to the corresponding amines followed by cleavage of amino group to release ammonia. Cell free extract showed nitroreductase activity in the range of 310-389 units/mg. NB was reduced to form benzamine and 4-aminophenol, 2NT was reduced to 2-aminotoluene, whereas 2NBA was reduced to form 2-aminobenzoic acid. Similarly, 3NT was metabolized to 3-aminotoluene and 2-amino-4-methylphenol, while 4NT was reduced to 4-nitrosotoluene and 4-aminotoluene. Cytotoxicity and apoptosis assays using Jurkat cell line, and Ames test were used to evaluate the detoxification of nitroaromatics during biodegradation. Biodegradation with Cupriavidus resulted in 2.6-11 fold increase in cell viability, 1.3-2.3 fold reduction in apoptosis, 1.6-55 fold reduction in caspase-3 activation, and complete disappearance of mutagenic activity. In soil microcosm, bioaugmentation with Cupriavidus resulted in 16-59% degradation of various nitroaromatics, as against <14% degradation without bioaugmentation. Thus, the present study reflects promising capability of Cupriavidus strain a3 in degradation and detoxification of multiple nitroaromatics., (Copyright © 2020 Elsevier Inc. All rights reserved.)

Soil management using fertilizers can modify soil chemical, biochemical and biological properties, including the concentration of trace-elements as cadmium (Cd), chromium (Cd) and nickel (Ni). Bacterial isolates from Cd, Cr, and Ni-contaminated soil were evaluated for some characteristics for their use in bioremediation. Isolates (592) were obtained from soil samples (19) of three areas used in three maize cultivation systems: no-tillage and conventional tillage with the application of mineral fertilizers; minimum tillage with the application of sewage sludge. Four isolates were resistant to Cr 3+ (3.06 mmol dm -3 ) and Cd 2+ (2.92 mmol dm -3 ). One isolate was resistant to the three metals at 0.95 mmol dm -3 . All isolates developed in a medium of Cd 2+ , Cr 3+ and Ni 2+ at 0.5 mmol dm -3 , and removed Cd 2+ (17-33%) and Cr 6+ (60-70%). They were identified by sequencing of the gene 16S rRNA, as bacteria of the genera Paenibacillus, Burkholderia, Ensifer, and two Cupriavidus. One of the Cupriavidus isolate was able to remove 60% of Cr 6+ from the culture medium and showed high indole acetic acid production capacity. We evaluated it in a microbe-plant system that could potentially be deployed in bioremediation by removing toxic metals from contaminated soil., (Published by Elsevier Inc.)