Nitro-polycyclic aromatic hydrocarbons (nitro-PAHs) are toxic and persistent environmental contaminants of increasing concern because of their widespread occurrence and ecological risks. Nitronaphthalene, a representative nitro-PAH, occurs as positional isomers, yet microbial degradation studies have mainly focused on 1-nitronaphthalene (1-NN), leaving the basis for biodegradation of 2-nitronaphthalene (2-NN) largely unresolved. In this study, Burkholderia pseudomultivorans M3 was isolated from river sediment in Anhui Province, China, and was found to efficiently degrade both 1-NN and 2-NN, indicating an expanded degradation range for nitronaphthalene isomers. Genome analysis identified a novel nitronaphthalene degradation gene cluster (nnd) that is phylogenetically related to reported nin- and nag-type gene clusters but differs functionally from both. Gene knockout and complementation experiments demonstrated that nndAc, encoding the catalytic α-subunit of a Rieske-type dioxygenase, is essential for degradation of both isomers. Heterologous expression and metabolite identification further showed that the NndAB system transforms 1-NN and 2-NN into 1,2-dihydroxynaphthalene via the corresponding nitro-dihydrodiol intermediates, with concomitant nitrite release. Site-directed mutagenesis revealed that the ability of NndAc depends on the coordinated effects of multiple active-site residues, with F200 being particularly important for 2-NN transformation. Moreover, strain M3 maintained substantial removal efficiency in synthetic wastewater containing mixed nitro-, carboxyl-, and hydroxyl-substituted naphthalene compounds, supporting its applicability in complex contaminated matrices. These findings reveal the molecular basis underlying the broadened substrate range of nitronaphthalene degradation and highlight the potential of strain M3 for remediation of wastewater containing mixed substituted naphthalene pollutants.
ABSTRACT Bisphenol F (BPF) is an emerging environmental pollutant widely present in surface water and wastewater systems. Microbial activity is crucial in driving its degradation, offering a potential avenue for mitigating its environmental impact. Although the degradation pathway for BPF has been identified in various bacteria, the biodegradation mechanisms remain unclear. In this study, we isolated a highly efficient BPF-degrading strain of Sphingobium yanoikuyae DN12, which could utilize BPF as the sole carbon and energy source for growth, from a river sediment in Anhui Province, China. Through ultra-performance liquid chromatography high-resolution mass spectrometry (UPLC-HRMS) analysis, we found that oxidation and hydrolysis are key steps for BPF biodegradation. Utilizing whole-genome sequencing, comparative transcriptomics analysis, and biochemical identification, a gene cluster bpf was identified to be involved in BPF degradation. BpfAB is a two-component oxidoreductase responsible for converting BPF to 4,4'-dihydroxybenzophenone (DHBP). BpfC is a Baeyer–Villiger monooxygenase responsible for converting DHBP to 4-hydroxyphenyl-4-hydroxybenzoate (HPHB). Isotope tracing demonstrated that the oxygen atom incorporated by BpfAB originates from water, whereas that incorporated by BpfC derives from molecular oxygen (O 2 ). BpfD is an α/β hydrolase responsible for converting HPHB to 4-hydroxybenzoate and 1,4-hydroquinone. Analysis of the taxonomic and habitat of 325 prokaryotic genomes revealed that BpfA-like homologs are predominantly found in the phylum Pseudomonadota , primarily inhabiting soil and aquatic environments. This study enhances our understanding of the biodegradation mechanism of BPF and provides guidance for the effective remediation of BPF-contaminated environments. IMPORTANCE Bisphenol F (BPF) is a widely used alternative to bisphenol A and poses a growing threat to ecosystems and human health due to its environmental persistence and endocrine-disrupting effects. Although microbial degradation pathways for BPF have been reported, the key enzymes involved and their catalytic mechanisms remain unclear. This work reports the isolation of a Sphingobium strain capable of mineralizing BPF and the genetic basis for the catabolic pathway. Three enzymes—a two-component oxidoreductase, a Baeyer–Villiger monooxygenase, and an α/β hydrolase—were biochemically characterized and shown to catalyze the three critical steps in BPF degradation. These findings provide insights into the biochemical processes involved in the microbial degradation of BPF.
ABSTRACT Gene redundancy endows bacteria with enhanced adaptability to complex and fluctuating environments but results in genetic costs. Transcriptional regulation is considered an effective strategy for harmonizing adaptive benefits with physiological burdens. In our previous study, two redundant gene clusters (phbh1pcaApcaBorf404bhbR1 and phbh2pcaB2pcaA2bhbR2) involved in 3-bromo-4-hydroxybenzoate (3-Br-4-HB) catabolism were identified in Pigmentiphaga kullae strain H8. The LysR-type transcription regulator BhbR1, encoded by the bhbR1 gene, activated phbh1pcaApcaBorf404 transcription. Through DNase I footprinting assays, the presence of the inducer 3-Br-4-HB was found to shorten the BhbR1-bound region in the promoter, uncovering the protected -35 box, thereby activating transcription. The MarR-family transcription factor (MFTF) BhbR2, encoded by the bhbR2 gene, was different from typical inhibitive MFTFs and activated phbh2pcaB2pcaA2 transcription. BhbR2 was found to bind a 17-bp imperfect palindromic sequence (TTGATT-N5-AATCAA) in the target promoter. Intriguingly, the presence of 3-Br-4-HB neither dissociated BhbR2 from the promoter nor modified its binding site, indicating a novel regulatory mode. Despite a coincident trend in activating their respective operons in response to different concentrations of 3-Br-4-HB, BhbR1 and BhbR2 both showed a significant attenuation of the activation effect at high concentrations (>480 μM), highlighting the requisite co-existence of redundant 3-Br-4-HB catabolic operons and their regulatory genes. This study presents two distinct transcriptional regulation mechanisms of these two redundant 3-Br-4-HB catabolic operons in strain H8, expanding our understanding of the diversity of transcriptional regulation for enhancing adaptation. IMPORTANCE In bacteria, catabolic genes for pollutant degradation often possess functionally redundant duplicates, providing a genetic basis for rapid adaptation to complex polluted environments. Synergic regulation plays an important role in balancing the physiological burden of extra genetic material with the adaptive benefits conferred by genetic redundancy. Although the co-existence of two redundant 3-bromo-4-hydroxybenzoate (3-Br-4-HB)-catabolic operons has been proven to enhance the metabolic robustness and adaptability of the host strain Pigmentiphaga kullae H8, how these two inducible catabolic operons are regulated remains unclear. This study identified two regulators, the LysR-type transcription regulator BhbR1 and the MarR-family transcription factor BhbR2, which activated transcription of the two 3-Br-4-HB-catabolic operons using different modes, and also revealed interactions of these two regulators with their effectors and target promoters. These findings not only clarify two distinct transcriptional strategies employed by redundant catabolic operons but also enhance our understanding of the significance of regulatory diversity for bacterial adaptation to complex polluted environments.
The implementation of environmentally friendly and sustainable remediation strategies positively impacts solid waste management. In this study, the Kocuria marina H-2 and Pseudomonas putida B6-2 co-culture system demonstrated enhanced naphthalene biodegradation efficiency compared to single-strain cultures. Under optimal conditions of 35 degrees C, 200 rpm/min, and a 1:1 ratio of the co-culture system, the naphthalene biodegradation potential was further increased. Notably, the addition of both ethylenediamine-pretreated lignin and phydroxybenzoic acid significantly elevated naphthalene degradation rates to 68.5 %. In addition, the oil-liquid surface tension decreased, while cell surface hydrophobicity and colony-forming units increased with the addition of lignin-derived compounds. The modification of naphthalene bioavailability by ethylenediaminepretreated lignin would accelerate the uptake and transport of hydrocarbons via ABC transporters and flagellar assembly. Importantly, genes related to bacterial chemotaxis and fatty acid biosynthesis were upregulated during the co-metabolism of naphthalene and p-hydroxybenzoic acid, further enhancing naphthalene bioconversion.
Nitrogen is a critical factor in plant growth, development, and crop yield. NODULE-INCEPTION-like proteins (NLPs), which are plant-specific transcription factors, function as nitrate sensors and play a vital role in the nitrogen response of plants. However, the genome-wide identification of the NLP gene family, the elucidation of the underlying molecular mechanism governing nitrogen response, and haplotype mining remain elusive in millet. In this study, we identified seven members of the NLP gene family in the millet genome and systematically analyzed their physicochemical properties. Evolutionary tree analysis indicated that SiNLP members can be classified into three subgroups, with NLP members from the same species preferentially grouped together within each subgroup. Analysis of gene structure characteristics revealed that all SiNLP members contained 10 conserved motifs, as well as the RWP-RK and PB1 domains, indicating that these motifs and domains have been relatively conserved throughout evolution. Additionally, we identified a significant abundance of response elements related to hormones, stress, growth, and development within the promoter regions of SiNLP members, suggesting that these members are involved in regulating diverse physiological processes in millet. Transcriptome data under low-nitrogen conditions showed significant differences in the expression profiles of SiNLP2 and SiNLP4 compared to the other members. RNA-seq and qRT-PCR results demonstrated that SiNLP2 significantly responds to low-nitrogen stress. Notably, we found that SiNLP2 is involved in nitrogen pathways by regulating the expression of the SiNAR2.1A, SiNAR2.1B, SiNRT1.1, and SiNR2 genes. More importantly, we identified an elite haplotype, Hap2, of SiNLP2, which is gradually being utilized in the breeding process. Our study established a foundation for a comprehensive understanding of the SiNLP gene family and provided gene resources for variety improvement and marker-assisted selection breeding.
Dissolved organic matter (DOM) is involved in numerous biogeochemical processes, and understanding the ecological succession of DOM is crucial for predicting its response to farming (e.g., fertilization) practices. Although plentiful studies have examined how fertilization practice affects the content of soil DOM, it remains unknown how long-term fertilization drives the succession of soil DOM over temporal scales. Here, we investigated the succession of DOM in paddy rice rhizosphere soils subjected to different long-term fertilization treatments (CK: no fertilization; NPK: inorganic fertilization; OM: organic fertilization) along with plant growth. Our results demonstrated that long-term fertilization significantly promoted the molecular chemodiversity of DOM, but it weakened the correlation between DOM composition and plant development. Time-decay analysis indicated that the DOM composition had a shorter halving time under CK treatment (94.7 days), compared to NPK (337.4 days) and OM (223.8 days) treatments, reflecting a lower molecular turnover rate of DOM under fertilization. Moreover, plant development significantly affected the assembly process of DOM only under CK, not under NPK and OM treatments. Taken together, our results demonstrated that long-term fertilization, especially inorganic fertilization, greatly weakens the ecological succession of DOM in the plant rhizosphere, which has a profound implication for understanding the complex plant-DOM interactions.
Dichlorprop [(R,S)-2-(2,4-dichlorophenoxy)propanoic acid], a chiral herbicide belonging to phenoxyalkanoic acid herbicides, is extensively used around the world, and is frequently detected in various habitats. Although (S)-dichlorprop has no herbicidal activity, its toxicity and ecological risk are higher than (R)-dichlorprop. In our previous studies, the isolated strain Sphingopyxis sp. DBS4 was confirmed to mineralize both (R)-dichlorprop and (S)-dichlorprop, but the key genes responsible for the initial degradation of (R)-/(S)-dichlorprop were still un-clear. Here, based on genome sequencing and enzyme purification, a new dioxygenase gene, spoA, responsible for the initial transformation of (S)-dichlorprop to 2,4-dichlorophenol and pyruvate was cloned from the rac- dichlorprop-utilizing strain of Sphingopyxis sp. DBS4. SpoA, an alpha-ketoglutarate-dependent dioxygenase, exhibited low amino acid sequence identity to the reported (S)-dichlorprop-transforming dioxygenase SdpA (34.8%), and showed enantioselective activity toward (S)-enantiomers of dichlorprop and mecoprop, but no activity toward the corresponding (R)-enantiomers. The Km value of SpoA for (S)-dichlorprop was 134.3 mu M, and the kcat/Km was determined to be 7.5 x 10-2 mu M-1 s- 1. Molecular docking and site-directed mutagenesis showed that two res-idues of SdpA, Lys101 and Asp119, are the key sites for the enantioselective transformation of (S)-dichlorprop. Our study identifies a new dioxygenase for the enantioselective transformation of (S)-dichlorprop and deepens our understanding of the microbial catabolism of chiral phenoxyalkanoic acid herbicides.
Wine can be regarded as a nutritional source for the human diet. It contains many nutrients such as vitamins and minerals, organic acids, flavonoids, and terpenoids. The varietal aroma of wines originated from the symbionts of the grapes and epiphytic microbiota, which combinedly grew from the sexual reproduction of the plant through seeds or during clonal reproduction. Nowadays, more and more studies focus on the assembly process of epiphytic microbiota and design a synthetic microbial community based on regional characteristics to improve wine quality and biofunctions. This review synthesizes the current concepts on the construction of synthetic microbiota, analyzes the advantages and difficulties in designing a functional yeast community, and lists the practical tools for data processing and model construction. A well-designed yeast community will possess high robustness against environment interference, higher efficiency of fermentation, and higher yield of targeted bioproducts.
Tetrabromobisphenol A (TBBPA), a widely used brominated flame retardant in electronics manufacturing, has caused global contamination due to improper e-waste disposal. Its persistence, bioaccumulation, and potential carcinogenicity drive studies of its transformation and underlying (a)biotic interactions. This study achieved an anaerobic enrichment culture capable of reductively dehalogenating TBBPA to the more bioavailable bisphenol A. 16S rRNA gene amplicon sequencing and quantitative PCR confirmed that successive dehalogenation of four bromide ions from TBBPA was coupled with the growth of both Dehalobacter sp. and Dehalococcoides sp. with growth yields of 5.0 ± 0.4 × 108 and 8.6 ± 4.6 × 108 cells per μmol Br- released (N = 3), respectively. TBBPA dehalogenation was facilitated by solid humin and reduced humin, which possessed the highest organic radical signal intensity and reducing groups -NH2, and maintained the highest dehalogenation rate and dehalogenator copies. Genome-centric metatranscriptomic analyses revealed upregulated putative TBBPA-dehalogenating rdhA (reductive dehalogenase) genes with humin amendment, cprA-like Dhb_rdhA1 gene in Dehalobacter species, and Dhc_rdhA1/Dhc_rdhA2 genes in Dehalococcoides species. The upregulated genes of lactate fermentation, de novo corrinoid biosynthesis, and extracellular electron transport in the humin amended treatment also stimulated TBBPA dehalogenation. This study provided a comprehensive understanding of humin-facilitated organohalide respiration.
Triclocarban, one of the emerging pollutants, has been accumulating, and it is frequently detected in wastewater. Due to its toxicity and persistence, the efficient removal of triclocarban from wastewater systems is challenging. Genetic bioaugmentation with transferable catabolic plasmids has been considered to be a long-lasting method to clean up pollutants in continuous flow wastewater treatment systems. In this study, bioaugmentation with Pseudomonas putida KT2440, harboring the transferrable triclocarban-catabolic plasmid pDCA-1-gfp-tccA2, rapidly converted 50 μM triclocarban in wastewater into 3,4-dichloroaniline and 4-chloroaniline, which are further mineralized more easily. RT-qPCR results showed that the ratio of the copy number of pDCA-1-gfp-tccA2 to the cell number of strain KT2440 gradually increased during genetic bioaugmentation, suggesting horizontal transfer and proliferation of the plasmid. By using DNA stable isotope probing (SIP) and amplicon sequencing, OTU86 (Escherichia-Shigella), OTU155 (Citrobacter), OTU5 (Brucella), and OTU15 (Enterobacteriaceae) were found to be the potential recipients of the plasmid pDCA-1-gfp-tccA2 in the wastewater bacterial community. Furthermore, three transconjugants in the genera of Escherichia, Citrobacter, and Brucella showing triclocarban-degrading abilities were isolated from the wastewater. This study develops a new method for removing triclocarban from wastewater and provides insights into the environmental behavior of transferrable catabolic plasmids in bacterial community in wastewater systems.
Genetic redundancy is prevalent in organisms and plays important roles in the evolution of biodiversity and adaptation to environmental perturbation. However, selective advantages of genetic redundancy in overcoming metabolic disturbance due to structural analogues have received little attention. Here, functional divergence of the three 4-hydroxybenzoate 3-hydroxylase (PHBH) genes (phbh1~3) was found in Pigmentiphaga sp. strain H8. The genes phbh1/phbh2 were responsible for 3-bromo-4-hydroxybenzoate (3-Br-4-HB, an anthropogenic pollutant) catabolism, whereas phbh3 was primarily responsible for 4-hydroxybenzoate (4-HB, a natural intermediate of lignin) catabolism. 3-Br-4-HB inhibited 4-HB catabolism by competitively binding PHBH3 and was toxic to strain H8 cells especially at high concentrations. The existence of phbh1/phbh2 not only enabled strain H8 to utilize 3-Br-4-HB but also ensured the catabolic safety of 4-HB. Molecular docking and site-directed mutagenesis analyses revealed that Val199 and Phe384 of PHBH1/PHBH2 were required for the hydroxylation activity towards 3-Br-4-HB. Phylogenetic analysis indicated that phbh1 and phbh2 originated from a common ancestor and evolved specifically in strain H8 to adapt to 3-Br-4-HB-contaminated habitats, whereas phbh3 evolved independently. This study deepens our understanding of selective advantages of genetic redundancy in prokaryote's metabolic robustness and reveals the factors driving the divergent evolution of redundant genes in adaptation to environmental perturbation.
With the aim of isolating clopyralid-degrading bacterial species for potential bioremediation, a pale-yellow, Gram-negative, rod-shaped, and non-motile designated as Clo-40T was isolated from soil which was about 10 years use of clopyralid in Zaozhuang city, Shandong province. Growth occurred within the ranges from 10 to 40 °C and 0–2.5% (w/v) NaCl. Strain could completely degrade 50 mg/L clopyralid within 2 days after induction and formed 3, 6-hydroxypicolinic acid, a major clopyralid metabolite, hydrolyze esculin, and reduce nitrates to nitrites, but could not hydrolyze gelatin. Based on phylogenetic analysis, strain clustered within the genus Xinfangfangia clade and branched with Xinfangfangia humi IMT-291T (97.6%) and Xinfangfangia soli ZQBWT (96.9%). Genome sequencing revealed a genome size of 4.41 Mbp and G + C content of 67.3%. The average nucleotide ANI values of strain with respect to X. humi IMT-291T and X. soli ZQBWT were 77.5% and 76.9%, respectively. The DDH estimated values between strain Clo-40T and X. humi IMT-291T and X. soli ZQBWT were 20.5% and 20.0%, respectively. The predominant fatty acids (> 5% of the total fatty acids) were C18:1 w7c (42.9%), C16:0 (28.8%), C17:0 cyclo (13.0%), and C14:0 (7.0%). The major polar lipids were identified as phosphatidylethanolamine, phosphatidylglycerol, unidentified phospholipid, unidentified glycolipid, and unidentified lipids. The predominant respiratory quinone was Q-10. Based on data from phenotypic, chemotaxonomic, and genotypic analyses in this study, strain Clo-40T represent a novel species in the genus of Xinfangfangia, for which the name Xinfangfangia pollutisoli sp. nov. is proposed. The type strain is Clo-40T (= KCTC 92089T = GDMCC 1.2845T).
Prokaryotes use various mechanisms, including improvement of the activity of detoxification enzymes, to cope with toxic intermediates produced during catabolism. However, studies on how bacteria accurately regulate differential transcriptions of various catabolic genes via a single regulator to ensure metabolic safety are scarce.
Plasmids disseminate pollutant-catabolic genes through horizontal transfer and contribute to the catabolic potential of the host microbial community. Genetic bioaugmentation of conjugative catabolic plasmids has recently been considered an effective and novel approach for long-term bioremediation of contaminated sites. In this study, we engineered an IncP-1β-2 subgroup plasmid pDCA-1 from Achromobacter sp. ANB-1 for its potential application in genetic bioaugmentation. A dcaA1A2B cluster responsible for the deamination of chloroaniline to chlorocatechol and a 1,2-dioxygenase gene (ccdC) for the ring cleavage of chlorocatechol were found to locate on the broad host range plasmid pDCA-1. Insertion of an amidase gene (phh or tccA2) at the accessory region of plasmid pDCA-1 greatly expanded its catabolic substrate spectrum from chloroaniline (3-chloroaniline and 3,4-dichloroaniline) to herbicides (linuron, propanil, propham, and chlorpropham), bacteriostatic agent (triclocarban), plant regulator (forchlorfenuron), and insecticide (diflubenzuron). Taking advantages of high-throughput cell sorting and 16S rRNA gene-based amplicon sequencing, we depicted the diversity of bacterial recipients for the engineered plasmid pDCA-1-gfp-phh in a soil bacterial consortium, showing an excellent conjugative transfer capacity of the pDCA-1 derivative to various Gram negative and even positive strains in natural environment. All in all, the engineered plasmid pDCA-1 had a great potential in genetic bioaugmentation of the sites contaminated with chloroanilines and their derivatives.
1-Naphthol, a widely used raw material for organic synthesis, is also a well-known organic pollutant. Due to its high toxicity, 1-naphthol is rarely used by microorganisms as the sole carbon source for growth. In this study, catabolism of 1-naphthol by Sphingobium sp. strain B2 was found to be greatly enhanced by additional supplementation with primary carbon sources (e.g., glucose, maltose, and sucrose), and 1-naphthol was even used as the carbon source for growth when strain B2 cells had been preinduced by both 1-naphthol and glucose. A distinct two-component flavin-dependent monooxygenase, NdcA1A2, was found to be responsible for the initial hydroxylation of 1-naphthol to 1,2-dihydroxynaphthalene, a more toxic compound. Transcriptional levels of ndcA1A2 genes were significantly upregulated when strain B2 cells were cultured with both 1-naphthol and glucose compared to cells cultured with only 1-naphthol or glucose. Two transcriptional regulators, the activator NdcS and the inhibitor NdcR, were found to play key roles in the synergistic regulation of the transcription of the 1-naphthol initial catabolism genes ndcA1A2IMPORTANCE Cometabolism is a widely observed phenomenon, especially in the field of microbial catabolism of highly toxic xenobiotics. However, the mechanisms of cometabolism are ambiguous, and the roles of the obligately coexisting growth substrates remain largely unknown. In this study, we revealed that the roles of the coexisting primary carbon sources (e.g., glucose) in the enhanced catabolism of the toxic compound 1-naphthol in Sphingobium sp. strain B2 were not solely because they were used as growth substrates to support cell growth but, more importantly, because they acted as coinducers to interact with two transcriptional regulators, the activator NdcS and the inhibitor NdcR, to synergistically regulate the transcription of the 1-naphthol initial catabolism genes ndcA1A2 Our findings provide new insights into the cometabolic mechanism of highly toxic compounds in microorganisms.
Strain CZZ-1T was isolated from long-term TBBPA-contaminated soil Zaozhuang city, Shandong province, People’s Republic of China. CZZ-1T was pink-pigmented, Gram-stain-negative, rod-shaped, non-motile and aerobic. The 16S rRNA gene analysis indicated that strain CZZ-1T shows high similarities to Fluviicola taffensis DSM 16823T (92.6%) and Fluviicola hefeinensis KACC 16597T (92.5%) and less than 91% sequence similarities to other genus or species in the family Crocinitomicaceae. It was able to grow at 10–37 °C, with 0–6% (w/v) NaCl. It could hydrolyze gelatin, but could not reduce nitrates to nitrites. The predominant fatty acids of strain CZZ-1T were iso-C15:0 (51.3%), C15:0 2-OH (11.0%), iso-C17:0 3-OH (8.0%), C14: 0 (7.0%), iso-C15:1 G (6.8%) and Summed Feature 3 (C16:1 ω7c and/or C16:1 ω6c, 4.4%). The polar lipid profile was composed of five unidentified lipids, two unidentified phospholipids, one phosphatidylethanolamine, one unidentified aminolipid and one unidentified glycolipid. The predominant respiratory quinone was MK-6. The genomic DNA G+C content of strain CZZ-1T was 41.5 mol%. Based on data from phenotypic, chemotaxonomic and genotypic analysis in this study, strain CZZ-1T represents a novel species in a new genus in the family Crocinitomicaceae, for which the name Taishania pollutisoli gen. nov., sp. nov. is proposed. The type strain is CZZ-1T (= KCTC 52343T = GDMCC 1.2270T).
The environmental fates of chlorinated 4-nitrophenols, 2,6-dichloro-4-nitrophenol (2,6-DCNP) and 2-chloro-4-nitrophenol (2C4NP), mediated via microbial catabolism have attracted great attention due to their high toxicity and persistence in the environment. In this study, a strain of Ensifer sp. 22-1 that was capable of degrading both 2,6-DCNP and 2C4NP was isolated from a halogenated aromatic-contaminated soil sample. A gene cluster cnpBADCERM was predicted to be involved in the catabolism of 2,6-DCNP and 2C4NP based on genome sequence analysis. A two-component monooxygenase CnpAB, composed of an oxygenase component (CnpA) and a reductase component (CnpB), was confirmed to catalyze the continuous denitration and dechlorination of 2,6-DCNP and 2C4NP to 6-chlorohydroxyquinol (6-CHQ) and hydroxyquinol (HQ), respectively. Knockout of cnpA resulted in the complete loss of the capacity for strain 22-1 to degrade 2,6-DCNP and 2C4NP. Homologous modeling and docking showed that Val155~Ala159, Phe206~Pro209 and Phe446~Arg461 of CnpA participated in the formation of the FAD-binding pocket, and Arg101, Val155 and Asn447 formed hydrogen bonds with 2,6-DCNP/2C4NP in the substrate-binding pocket. This work characterized a new two-component monooxygenase for 2,6-DCNP and 2C4NP, and enriched our understanding of the degradation mechanism of chlorinated nitrophenols (CNPs) by microorganisms.
肼是一种广泛使用的化工原料,但它也是一种有毒化学品,对人类健康和环境安全有着严重威胁.因此,开发一种方便、快速检测肼的方法具有重要的意义.本文制备了一种芘甲醛纳米粒子探针,其能和肼快速反应,从而使探针的荧光信号发生变化,实现对肼的荧光检测.该探针检测肼具有高选择性和灵敏度,并成功地应用于HeLa细胞和斑马鱼中肼的成像.
Atrazine, a triazine herbicide, is widely used around the world. The residue of atrazine due to its application in the fore-rotating crop maize has caused phytotoxicity to the following crop sweet potato in China. Bioaugmentation of atrazine-contaminated soil with atrazine-degrading strains is considered as the most potential method to remove atrazine from soil. Nevertheless, the feasibility of bioaugmentation and its effect on soil microbiome still need investigation. In this study, Paenarthrobacter sp. AT-5, an atrazine-degrading strain, was inoculated into agricultural soils contaminated with atrazine to investigate the bioaugmentation process and the reassembly of the soil microbiome. It was found that 95.9% of 5 mg kg−1 atrazine was removed from the soils when inoculated with strain AT-5 with 7 days, and the phytotoxicity of sweet potato caused by atrazine was significantly alleviated. qRT-PCR analysis revealed that the inoculated strain AT-5 survived well in the soils and maintained a relatively high abundance. The inoculation of strain AT-5 significantly affected the community structure of the soil microbiome, and the abundances of bacteria associated with atrazine degradation were improved.