The exposure risks associated with pathogens and antibiotic resistance genes in bioaerosol from municipal landfill and surrounding area.

Pathogenic microbes with antibiotic resistance can thrive on municipal solid waste as nutrients and be aerosolized and transported to vicinities during waste disposal processes. However, the characterization of pathogenic bioaerosols and assessment of their exposure risks are lacking. Herein, particle size, concentration, activity, antibiotic resistance, and pathogenicity of airborne microorganisms were assessed in different sectors of a typical landfill. Results showed that active sector in downwind direction has the highest bioaerosol level (1234 CFU/m3), while residential area has the highest activity (14.82 mg/L). Botanical deodorizer from mist cannon can effectively remove bioaerosol. Most bioaerosols can be inhaled into respiratory system till bronchi with sizes ranging from 2.1-3.3 and 3.3-4.7 µm. Pathogenic bacteria (Bacilli, Bacillus, and Burkholderia-Paraburkholderia) and allergenic fungi (Aspergillus, Cladosporium, and Curvularia) prevailed in landfill. Although high abundance of microbial volatile organic compounds (mVOCs) producing bioaerosols were detected, these mVOCs contributed little to odor issues in landfill. Notably, surrounding areas have higher levels of antibiotic-resistance genes (ARGs) than inner landfill with tetC, acrB, acrF, mdtF, and bacA as dominant ones. Most ARGs were significantly correlated with bacterial community, while environmental parameters mainly influenced fungal prevalence. These findings can assist in reducing and preventing respiratory allergy or infection risks in occupational environments relating to waste management.

Genome sequence of a spore-laccase forming, BPA-degrading Bacillus sp. GZB isolated from an electronic-waste recycling site reveals insights into BPA degradation pathways.

Bisphenol A (BPA) is a synthetic chemical with known deleterious effects on biota. A genome sequencing project is an important starting point for designing a suitable BPA bioremediation process, because it provides valuable genomic information about the physiological, metabolic, and genetic potential of the microbes used for the treatment. This study explored genomic insights provided by the BPA-degrading strain Bacillus sp. GZB, previously isolated from electronic-waste-dismantling site. The GZB genome is a circular chromosome, comprised of a total of 4,077,007 bp with G+C content comprising 46.2%. Genome contained 23 contigs encoded by 3881 protein-coding genes with nine rRNA and 53 tRNA genes. A comparative study demonstrated that strain GZB bloomed with some potential features as compared to other Bacillus species. In addition, strain GZB developed spore cells and displayed laccase activity while growing at elevated stress levels. Most importantly, strain GZB contained many protein-coding genes associated with BPA degradation, as well as the degradation of several other compounds. The protein-coding genes in the genome revealed the genetic mechanisms associated with the BPA degradation by strain GZB. This study predicts four possible degradation pathways for BPA, contributing to the possible use of strain GZB to remediate different polluted environments in the future.

Indoor/Outdoor airborne microbiome characteristics in residential areas across four seasons and its indoor purification.

Bioaerosols are more likely to accumulate in the residential environment, and long-term inhalation may lead to a variety of diseases and allergies. Here, we studied the distribution, influencing factors and diffusion characteristics of indoor and outdoor microbiota pollution in six residential buildings in Guangzhou, southern China over a period of one year. The results showed that the particle sizes of bioaerosol were mainly in the range of inhalable particle size (<4.7 µm) with a small difference among four seasons (74.61 % ± 2.17 %). The microbial communities showed obvious seasonal differences with high abundance in summer, but no obvious geographical differences. Among them, the bacteria were more abundant than the fungi. The dominant microbes in indoor and outdoor environments were similar, with Anoxybacillu, Brevibacillus and Acinetobacter as the dominant bacteria, and Cladosporium, Penicillium and Alternaria as the dominant fungi. The airborne microbiomes were more sensitive to temperature and particulate matter (PM2.5, PM10) concentrations. Based on the Sloan neutral model, bacteria were more prone to random diffusion than fungi, and the airborne microbiome can be randomly distributed in indoor and outdoor environments and between the two environments in each season. Bioaerosol in indoor was mainly from outdoor. The health risk evaluation showed that the indoor inhalation risks were higher than those outdoor. The air purifier had a better removal efficiency on 1.1-4.7 µm microorganisms, and the removal efficiency on Gram-negative bacteria was better than that on Gram-positive bacteria. This study is of great significance for the risk assessment and control of residential indoor bioaerosol exposure.

Recent progress in online detection methods of bioaerosols.

The aerosol transmission of coronavirus disease in 2019, along with the spread of other respiratory diseases, caused significant loss of life and property; it impressed upon us the importance of real-time bioaerosol detection. The complexity, diversity, and large spatiotemporal variability of bioaerosols and their external/internal mixing with abiotic components pose challenges for effective online bioaerosol monitoring. Traditional methods focus on directly capturing bioaerosols before subsequent time-consuming laboratory analysis such as culture-based methods, preventing the high-resolution time-based characteristics necessary for an online approach. Through a comprehensive literature assessment, this review highlights and discusses the most commonly used real-time bioaerosol monitoring techniques and the associated commercially available monitors. Methods applied in online bioaerosol monitoring, including adenosine triphosphate bioluminescence, laser/light-induced fluorescence spectroscopy, Raman spectroscopy, and bioaerosol mass spectrometry are summarized. The working principles, characteristics, sensitivities, and efficiencies of these real-time detection methods are compared to understand their responses to known particle types and to contrast their differences. Approaches developed to analyze the substantial data sets obtained by these instruments and to overcome the limitations of current real-time bioaerosol monitoring technologies are also introduced. Finally, an outlook is proposed for future instrumentation indicating a need for highly revolutionized bioaerosol detection technologies.

Online monitoring system for qualitative and quantitative analysis of bioaerosols by combined ATP bioluminescence assay with loop-mediated isothermal amplification.

Rapid detection of airborne pathogens is crucial in preventing respiratory infections and allergies. However, technologies aiming to real-time analysis of microorganisms in air remain limited due to the sparse and complex nature of bioaerosols. Here, we introduced an online bioaerosol monitoring system (OBMS) comprised of integrated units including a rotatable stainless-steel sintered filter-based sampler, a lysis unit for extracting adenosine triphosphate (ATP), and a single photon detector-based fluorescence unit. Through optimization of the ATP bioluminescence method and establishment of standard curves between relative luminescence units (RLUs) and ATP as well as microbial concentration, we achieved simultaneous detection of bioaerosols' concentration and activity. Testing OBMS with four bacterial and two fungal aerosols at a sampling flow rate of 10 to 50 L/min revealed an outstanding collection efficiency of 95 % at 30 L/min. A single OBMS measurement takes only 8 min (sampling: 5 min; lysis and detection: 3 min) with detection limits of 3 Pcs/ms photons (2.9 × 103 and 292 CFU/m3 for Staphylococcus aureus and Candida albicans aerosol). In both laboratory and field tests, OBMS detected higher concentrations of bioaerosol compared to the traditional Andersen impactor and liquid biosampler. When combined OBMS with loop-mediated isothermal amplification (LAMP), the bioaerosol can be qualitative and quantitative analyzed within 40 min without the cumbersome procedures of sample pretreatment and DNA extraction. These results offer a high compressive and humidity resistance membrane filtration sampler and validate the potential of OBMS for online measurement of bioaerosol concentration and composition.

The abundance and pathogenicity of microbes in automobile air conditioning filters across the typical cities of China and Europe.

Bioaerosols are widely distributed in urban air and can be transmitted across the atmosphere, biosphere, and anthroposphere, resulting in infectious diseases. Automobile air conditioning (AAC) filters can trap airborne microbes. In this study, AAC filters were used to investigate the abundance and pathogenicity of airborne microorganisms in typical Chinese and European cities. Culturable bacteria and fungi concentrations were determined using microbial culturing. High-throughput sequencing was employed to analyze microbial community structures. The levels of culturable bioaerosols in Chinese and European cities exhibited disparities (Analysis of Variance, P < 0.01). The most dominant pathogenic bacteria and fungi were similar in Chinese (Mycobacterium: 18.2-18.9 %; Cladosporium: 23.0-30.2 %) and European cities (Mycobacterium: 15.4-37.7 %; Cladosporium: 18.1-29.3 %). Bartonella, Bordetella, Alternaria, and Aspergillus were also widely identified. BugBase analysis showed that microbiomes in China exhibited higher abundances of mobile genetic elements (MGEs) and biofilm formation capacity than those in Europe, indicating higher health risks. Through co-occurrence network analysis, heavy metals such as zinc were found to correlate with microorganism abundance; most bacteria were inversely associated, while fungi exhibited greater tolerance, indicating that heavy metals affect the growth and reproduction of bioaerosol microorganisms. This study elucidates the influence of social and environmental factors on shaping microbial community structures, offering practical insights for preventing and controlling regional bioaerosol pollution.

Bioaerosols in an industrial park and the adjacent houses: Dispersal between indoor/outdoor, the impact of air purifier, and health risk reduction.

Inhaling airborne pathogens may cause severe epidemics showing huge threats to indoor dwellings residents. The ventilation, environmental parameters, and human activities would affect the abundance and pathogenicity of bioaerosols in indoor. However, people know little about the indoor airborne microbes especially pathogens near the industrial park polluted with organics and heavy metals. Herein, the indoor bioaerosols' community composition, source and influencing factors near an electronic waste (e-waste) industrial park were investigated. Results showed that the average bioaerosol level in the morning was lower than evening. Bioaerosol concentration and activity in indoor (1936 CFU/m3 and 7.62 × 105 ng/m3 sodium fluorescein in average) were lower than the industrial park (4043 CFU/m3 and 7.77 × 105 ng/m3 sodium fluorescein), and higher microbial viability may be caused by other pollutants generated during e-waste dismantling process. Fluorescent biological aerosol particles occupied 17.6%-23.7% of total particles, indicating that most particles were non-biological. Bacterial communities were richer and more diverse than fungi. Furthermore, Bacillus and Cladosporium were the dominant indoor pathogens, and pathogenic fungi were more influenced by environmental factors than bacteria. SourceTracker analysis indicates that outdoor was the main source of indoor bioaerosols. The hazard quotient (<1) of airborne microbes through inhalation was negligible, but long-term exposure to pathogens could be harmful. Air purifiers could effectively remove the airborne fungi and spheroid bacteria than cylindrical bacteria, but open doors and windows would reduce the purification efficiency. This study is great important for risk assessments and control of indoor bioaerosols near industrial park.

BPA degradation using biogenic manganese oxides produced by an engineered Escherichia coli with a non-blue laccase from Bacillus sp. GZB.

Bisphenol A (BPA), an endocrine disruptor that is often found in a variety of environmental matrixes, poses a serious health risk. One of the most effective methods for completely degrading BPA is biological oxidation. This study used a non-blue laccase to develop an engineer Escherichia coli strain for the synthesis of biogenic manganese oxides (BMO). The recombinant strain LACREC3 was utilized for the efficient production of BMO. The LACREC3 strain developed the erratic clumps of BMO after prolonged growth with Mn2+, as shown by scanning electron microscopy (SEM) and energy-dispersive X-ray (EDS) tests. After 12 days of incubation under liquid culture conditions, a total of 51.97 ± 0.56% Mn-oxides were detected. The Brunauer-Emmett-Teller (BET) surface areas, X-ray diffraction (XRD), Fourier transform infrared (FT-IR), and X-ray photoelectron spectroscopy (XPS) experiments were further used to characterize the purified BMO. Data revealed that Mn(IV)-oxides predominated in the structure of BMO, which was amorphous and weakly crystalline. The BPA oxidation assay confirmed the high oxidation efficiency of BMO particle. BMO degraded 96.16 ± 0.31% of BPA in total over the course of 60 min. The gas chromatography and mass spectroscopy (GC-MS) identified BPA-intermediates showed that BPA might break down into less hazardous substances that were tested by Photobacterium Phosphoreum in an acute toxicity experiment. Thus, employing BMO generated by a non-blue laccase, this study introduces a new biological technique of metal-oxidation and organic-pollutant degradation.

Bioinspired artificial spider silk photocatalyst for the high-efficiency capture and inactivation of bacteria aerosols.

Bioaerosol can cause the spread of disease, and therefore, capture and inactivation of bioaerosols is desirable. However, filtration systems can easily become blocked, and are often unable to inactivate the bioaerosol once it is captured. Herein, we reported a bioinspired artificial spider silk (ASS) photocatalyst, consisting of a periodic spindle structure of TiO2 on nylon fiber that can efficiently capture and concentrate airborne bacteria, followed by photocatalytic inactivation in situ, without a power-supply exhaust system. The ASS photocatalyst exhibits a higher capture capacity than the nylon fiber substrate and a photocatalytic inactivation efficiency of 99.99% obtained under 4 h irradiation. We found that the capture capacity of the ASS photocatalyst can be mainly attributed to the synergistic effects of hydrophilicity, Laplace pressure differences caused by the size of the spindle knots and surface energy gradients induced by surface roughness. The bacteria captured by the ASS photocatalyst are inactivated by photocatalysis within droplets or at the air/photocatalyst interfaces. This strategy paves the way for constructing materials for bioaerosol purification.

Metagenomic profiles and health risks of pathogens and antibiotic resistance genes in various industrial wastewaters and the associated receiving surface water.

The aquatic environment may represent an essential route for transmission of antibiotic resistance to opportunistic human pathogens. Since industrial wastewater is discharged into the river after treatment, understanding the distribution of antibiotic resistance genes (ARGs) in river systems and the possibility of pathogens acquiring antibiotic resistance are challenges with far-reaching significance. This work mainly studied distribution profiles of pathogens and ARGs, and compared their health risk in various industrial wastewater with that of river water. Results showed that 166 pathogens were concurrently shared by the six water samples, with Salmonella enterica and Pseudomonas aeruginosa being the most abundant, followed by Fusarium graminearum and Magnaporthe oryzae. The similar composition of the pathogens suggests that pathogens in river water may mainly come from sewage discharge of slaughterhouses and that changes in water quality contribute significantly to the prevalence of these pathogens. Of the 57 ARG types detected, bacitracin was the most abundant, followed by sulfonamide, chloramphenicol, tetracycline, and aminoglycoside. Strikingly, the wastewater from a pharmaceutical factory producing Chinese medicine was also rich in bacA, sul1, mexW, mexB, mexF and oprn. It can be seen from the co-occurrence patterns that pathogens and the main ARGs have strong co-occurrence. Higher abundance of offensive virulence factors in industrial wastewater and their strong correlation with pathogens containing ARGs suggest higher microbiological risk. These findings highlight the need to assess ARG acquisition by pathogens in the surface water of human-impacted environments where pathogens and ARGs may co-thrive.

Contributions of meat waste decomposition to the abundance and diversity of pathogens and antibiotic-resistance genes in the atmosphere.

Airborne transmission of antibiotic-resistance genes (ARGs) in landfill and acquisition of antibiotic resistance by pathogenic bacteria are posing potential threat to human and environmental health. However, little is known about contribution of waste decomposition to airborne ARGs and pathogens during landfilling of household waste. Herein, the dynamic changes of microbial communities and ARGs were comparatively investigated in leachate and bioaerosol during the decomposition of chicken, fish, and pork wastes. Results found that chicken and pork decomposition could result in emitting high abundance of bioaerosol and pathogen, while fish fermentation will lead to high airborne microbial activity. The main pathogens were Bacilli, Burkholderia-Paraburkholderia and Mycobacterium in bioaerosols, but were Wohlfahrtiimonas, Peptoniphilus and Fusobacterium in leachate, suggesting that the ability of aerosolization of bacteria in leachate was independent of their abundance and diversity. Whereas, diversity and relative abundance of ARGs in leachate were significantly higher than bioaerosol. Moreover, the relative abundance of ARGs in leachate and bioaerosols was not completely relevant. The changes of pathogenic community contributed significantly to the prevalence of ARGs in bioaerosol and leachate. The results will define the contribution of household waste decomposition to airborne pathogen and ARG distribution and provide foundation for airborne bacterial exposure risk and control in landfill.

A non-blue laccase of Bacillus sp. GZB displays manganese-oxidase activity: A study of laccase characterization, Mn(II) oxidation and prediction of Mn(II) oxidation mechanism.

Laccase, a unique class of multicopper oxidase, presents promising potential as a biocatalyst in many industrial and biotechnological applications. Recently, it has been significantly applied in many metal-polluted sites due to its Manganese (Mn)-oxidation ability. Here, we demonstrate the Mn(II)-oxidase activity of laccase obtained from Bacillus sp. GZB. The CotA gene of GZB was transformed in E. coli BL21 and overexpressed. The purified laccase (LACREC3-laccase) displayed the absence of a peak at 610 nm that is usually found in blue-laccase. Further, the LACREC3-laccase exhibited high activity and stability at different pH and temperatures with substrates 2, 2'-Azino-bis (3-ethylbenzothiazoline-6-sulfonate) and syringaldazine, respectively. It also functioned in the presence of various metals and enzyme inhibitors. Most notably, LACREC3-laccase formed insoluble brown Mn(III)/Mn(IV)-oxide particles from Mn(II) mineral, exhibiting its Mn(II)-oxidase activity. In addition to native polyacrylamide gel electrophoresis and buffer test, we developed an 'agarose gel plate' assay to evaluate Mn(II) oxidation activity of laccase. Furthermore, using the leucoberbelin blue assay, a total of 44.45 ± 0.45% Mn(IV)-oxides were quantified, in which 5.87 ± 0.61% autoxidized after 24 h. The Mn(II) oxidation mechanisms were further predicted by trapping Mn(III) using pyrophosphate during Mn(II) to Mn(IV) conversion by LACREC3-laccase. Overall, the laccase of GZB has excellent activity and stability plus an ability to oxidize Mn(II). This study is the first report on a non-blue laccase, exhibiting Mn(II)-oxidase activity. Thus, it offers a novel finding of the Mn(II) oxidation processes that can be a valuable way of Mn(II)-mineralization in various metal-polluted environments.

Pollution profiles of antibiotic resistance genes associated with airborne opportunistic pathogens from typical area, Pearl River Estuary and their exposure risk to human.

To reveal the selective pressures of near-shore human activities on marine and continental bioaerosols, the pollution profile and potential exposure risk of airborne pathogens and antibiotic-resistance genes (ARGs) in Pearl River Estuaries (113.52 oE, 22.69 oN), a transitional zone between marine and continental environments, were fully explored. The results showed that the total bacteria among bioaerosols varied largely with average pollution levels of 1.86 × 105 and 4.35 × 104 cfu m-3 in spring and summer, respectively, and were high than those of airborne fungi. The predominant aerodynamic diameters of bioaerosols were in respirable size range (<4.7 µm), and the microbes communities' diversity and abundance varied significantly. Besides, many opportunistic pathogenic bacteria (Burkholderia-Paraburkholderia, Staphylococcus and Acinetobacter) and fungi (Alternaria, Penicillium and Cladosporium) were dominant in bioaerosol samples. Of 21 ARGs subtypes detected, the tetracycline resistance gene tetA was the most abundant, followed by aminoglycoside resistance gene and mobile genetic elements. Correlation analysis revealed that the changes of pathogens community contributed significantly to the prevalence of ARGs in bioaerosol. Based on the average daily dose rates of microorganisms and human direct intake of ARGs, health risk of bioaerosols from the Pearl River Estuaries were also evaluated. In summary, the presence of opportunistic pathogens and diversity of ARGs strengthens the call to consider the bioaerosol in air quality monitoring and risk assessment in the future.

The formation mechanism of antibiotic-resistance genes associated with bacterial communities during biological decomposition of household garbage.

Food wastes are significant reservoir of antibiotic-resistance genes (ARGs) and antibiotic-resistant bacteria (ARB) available for exchange with clinical pathogens. However, food wastes-related changes of antibiotic resistance in long-period decomposition have been overlooked. Here, we evaluated the comprehensive ARG profile and its association with microbial communities, explained how this might vary with household garbage decomposition. Average of 128, 150 and 91 ARGs were detected in meat, vegetable and fruit wastes, respectively, with multidrug and tetracycline as the predominant ARG types. ARG abundance significantly increased at initial stage of waste fermentation and then decreased. High abundance of Eubacterium-coprostanoligenes, Sporanaerobacter, Peptoniphilus, Peptostreptococcus might be explained for the high relative abundance of ARGs in meat, while high abundance of Advenella, Prevotella, Solobacterium was attributed to the high diversity of ARGs in vegetables. Significant correlations were observed among volatile organic compounds, mobile genetic elements and ARGs, implying that they might contribute to transfer and transport of ARGs. Network analysis revealed that aph(2')-Id-01, acrA-05, tetO-1 were potential ARG indicators, while Hathewaya, Paraclostridium and Prevotellaceae were possible hosts of ARGs. Our work might unveil underlining mechanism of the effects of food wastes decomposition on development and spread of ARGs in environment and also clues to ARG mitigation.

Malodorous gases production from food wastes decomposition by indigenous microorganisms.

Volatile organic compounds (VOCs) produced during the degradation of food wastes may harm to the health of people and create annoyance in adjacent communities. In this work, the VOCs emitted from the decomposition food wastes including fruit, meat and vegetable, and their microbial communities were measured in three individual 57-L reactors for 61 days. Total of 232.8, 373.5, and 191.1 µg·kg-1·h-1 VOCs with oxygenated VOCs (57.6%), volatile organic sulfur compounds (VOSCs, 58.6%) and VOSCs (54.9%) as the main group were detected during fruit, meat and vegetable fermentation, respectively. 2-Butanone (55.1%) and ethyl acetate (13.8%) were the two most abundant VOCs from fruit wastes, while dimethyl sulfide (68.0 and 26.6%) and dimethyl disulfide (89.2 and 10.1%) were in vegetable and meat wastes. The predominant Firmicutes represented 93.0-99.9% of the bacterial communities of meat decomposition, while Firmicutes and Proteobacteria were the dominant phyla throughout the fruit digestion process. Proteobacteria (16.9%-83.6%) was the dominant phylum in vegetable wastes, followed by Bacteroidetes, Firmicutes, and Actinobacteria. Malodorous VOCs emissions were highly affected by microbial activity, the abundant Weissella, Leuconostoc and Enterobacteriaceae in vegetable wastes showed correlation with carbon disulfide and dimethyl sulfide, while dominant Peptococcus, Bacteroides, Lactobacillales and Peptoniphilus in meat wastes was related to dimethyl disulfide. Overall, significant differences and correlation between VOCs emission profiles and bacterial communities among different food wastes decomposition were observed. These data contribute to a more comprehensive understanding the relationship between microbial community dynamics and malodorous VOCs emission.

Biodegradation of typical BFRs 2,4,6-tribromophenol by an indigenous strain Bacillus sp. GZT isolated from e-waste dismantling area through functional heterologous expression.

Legacy wastewater contaminants from e-waste dismantling process such as 2,4,6-tribromophenol (TBP), one of the most widely used brominated flame retardants (BFRs), have raised concern owing to their toxicity and recalcitrance. Our previously isolated Bacillus sp. GZT from river sludge in e-waste dismantling area is a good candidate for bioremediation of BFRs contaminated sites considering its remarkable degradability of TBP and its intermediates. However, there exists a new challenge because bio-degrader cannot produce enough biomass or metabolic activity to cleanup TBP in practice complex environment. Here, we heterologously expressed and functionally characterized the genes and enzymes responsible for TBP degradation to examine the feasibility of enhancing the ability of this microorganism to detoxify TBP. Results demonstrated that five recombinant strains containing functional genes, designated tbpA, tbpB, tbpC, tbpD, and tbpE, become more tolerant toward a wide range of brominated compounds than the nontransgenic strain. Cytochrome P450 reductase encoded by tbpA gene could greatly increase efficiency to remove TBP (98.8%), as compared to wild-type strain GZT (93.2%). Its debromination intermediates 2,4-dibromophenol, 2,6-dibromo-4-methylphenol and 2-bromophenol were significantly metabolized by halophenol dehalogenases encoded by tbpB, tbpC, and tbpD, respectively. Finally, under the function of tbpE gene encoding enzyme, further debrominated product (phenol) was dramatically detoxified. To reduce the risk of these xenobiotics, the expression of these genes can be induced and significantly up-regulated during exposure to them. These results open broad scope for future study in developing genetic engineering technologies for more efficient remediation wastewater of e-waste recycling sites contaminated with TBP, which would certainly be important steps to lower TBP exposures and prevent potential health effects.

Application of a novel gene encoding bromophenol dehalogenase from Ochrobactrum sp. T in TBBPA degradation.

Tetrabromobisphenol-A (TBBPA), a typical brominated flame retardant, leaked from commercial products into the environments has attracted people's attention around the world. Ochrobactrum sp. T capable of degradation and mineralization of TBBPA was isolated in our early work. In this study, the identification of TBBPA-degrading gene from the strain was further carried out by combining whole-genome sequencing with gene cloning and expression procedures. In total, 3877 open reading frames were found within 3.9 Mb genome and seven of them were identified as dehalogenating-relating genes. One gene with a significant ability to degrade TBBPA was designated as tbbpaA. Sequence alignments analysis showed that it shared 100% identity with haloacid dehalogenases. Furthermore, tbbpaA gene was cloned and expressed into E. coli to achieve a constructed strain. Like the original strain, the constructed strain could degrade TBBPA (6 mg L-1) with 78% of debromination efficiency and 37.8% mineralization efficiency within 96 h. Gene expression study revealed that tbbpaA was up-regulated in the presence of TBBPA. Overall, we report the identification of a functional TBBPA-degrading gene in an aerobe, which can deepen the knowledge of enhancing TBBPA removal by Strain T at the genetic level and facilitate in situ TBBPA bioremediation.

Purification, molecular characterization and metabolic mechanism of an aerobic tetrabromobisphenol A dehalogenase, a key enzyme of halorespiration in Ochrobactrum sp. T.

Due to the detoxification of tetrabromobisphenol A (TBBPA) varies from different bacterial strains and depends on their specific enzymatic machinery, it is necessary to understand them for potential in situ bioremediation application. The special ability of our previously isolated Ochrobactrum sp. T to simultaneously debrominate and aerobic mineralize TBBPA urgent us to continuously study its degradation molecular mechanism. Herein, the purification and characterization of the dehalogenase which can debrominate TBBPA was investigated based on its corresponding encoding gene tbbpaA. Results showed that an enzyme with molecular mass of 117 kDa, Km of 26.6 µM and Vmax of 0.133 µM min-1 mg-1 was purified and designated as bromophenol dehalogenase. It was the only detected dehalogenase which exhibited TBBPA degradation ability (78%). Moreover, its activity was significantly enhanced by adding NADPH or methyl viologen to the reaction solution. The high similarity of substrate spectrum between the dehalogenase from the recombinant strain and the wild strain further indicated that it was the main dehalogenase responsible for the debromination in wild strain. Based on three identified metabolites, a metabolic pathway of TBBPA by purified enzyme under oxic condition was proposed. This study provides an excellent dehalogenase candidate for mechanistic study of aerobic dehalogenation of brominated aromatic compound.

Purifying, cloning and characterizing a novel dehalogenase from Bacillus sp. GZT to enhance the biodegradation of 2,4,6-tribromophenol in water.

2,4,6-Tribromophenol (TBP), an intermediate of brominated flame retardants, can easily release to environment and recalcitrant to degradation. Previously, Bacillus sp. GZT, a pure aerobic strain capable of simultaneously debrominating and mineralizing TBP, was successfully isolated by us. To further obtain a practical application and dig up its TBP degradation mechanism, a total of 46.7-fold purification of a novel dehalogenase with a final specific activity of 18.9 U mg-1 and a molecular mass of 63.4 kDa was achieved. Under optimal conditions (35 °C and 200 rpm), up to 80% degradation efficiencies were achieved within 120 min. Adding H2O2, NADPH, Mn2+ and Mg2+ promoted enzyme reaction effectively; while EDTA, methyl viologen, Ni2+, Cu2+, Ca2+ and Fe2+ strongly inhibited reaction activities. The debromination of TBP was catalyzed by the enzyme at a Km of 78 µM and a Vmax of 0.65 min-1 mg protein-1, which indicated that this dehalogenase could specifically eliminate TBP with a high efficiency and stability. Based on MALDI-TOF/TOF analysis, the dehalogenase shared 98% identity with peptide ABC transporter substrate-binding protein. One open reading frame (ORF) encoding this peptide was found in Strain GZT genome, subjected to clone and expressed in Escherichia coli (E. coli) to characterize the encoding gene. Result showed that this recombinant strain could also remove as similar amount of TBP as Bacillus sp. GZT under the identical condition. Based on these results, we suggest that this newly-isolated TBP dehalogenase highlights a new approach for remediating TBP pollution.

Draft Genome Sequence of Bacillus sp. GZT, a 2,4,6-Tribromophenol-Degrading Strain Isolated from the River Sludge of an Electronic Waste-Dismantling Region.

Here, we report the draft genome sequence of Bacillus sp. strain GZT, a 2,4,6-tribromophenol (TBP)-degrading bacterium previously isolated from an electronic waste-dismantling region. The draft genome sequence is 5.18 Mb and has a G+C content of 35.1%. This is the first genome report of a brominated flame retardant-degrading strain.