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Bioresource Technology Feb 2024Heterotrophic nitrification-aerobic denitrification (HN-AD) shows innovation potential of wastewater treatment process in a single tank. However, how to enrich HN-AD...
Feast/famine ratio regulates the succession of heterotrophic nitrification-aerobic denitrification and autotrophic ammonia oxidizing bacteria in halophilic aerobic granular sludge treating saline wastewater.
Heterotrophic nitrification-aerobic denitrification (HN-AD) shows innovation potential of wastewater treatment process in a single tank. However, how to enrich HN-AD bacteria in activated sludge to enhance their contribution remained unknown. This study explored the impact of the feast/famine (F/F) ratio on the succession of autotrophic ammonia oxidizing bacteria (AOB) and HN-AD bacteria in a halophilic aerobic granular sludge (HAGS) system. As the F/F ratio decreased from 1/9 to 1/15, the total inorganic nitrogen (TIN) removal performance significantly decreased. The proportion of heterotrophic bacteria was dropped from 79.0 % to 33 %. Accordingly, the relative abundance of Paracoccus decreased from 70.8 % to 25.4 %, and the copy number of the napA gene was reduced from 2.2 × 10 copies/g HAGS to 8.1 × 10 copies/g HAGS. It found the F/F ratio regulated the population succession of autotrophic AOB and HN-AD bacteria, thereby providing a solution to achieve the enrichment of HN-AD bacteria in HAGS.
Topics: Nitrification; Wastewater; Sewage; Denitrification; Ammonia; Bioreactors; Heterotrophic Processes; Bacteria; Nitrogen; Betaproteobacteria; Oxidation-Reduction; Aerobiosis
PubMed: 37951552
DOI: 10.1016/j.biortech.2023.129995 -
Bioresource Technology Feb 2024L-theanine is a natural non-protein amino acid with wide applications. Thus, a high yield of L-theanine production is required on an industrial scale. Herein, an...
L-theanine is a natural non-protein amino acid with wide applications. Thus, a high yield of L-theanine production is required on an industrial scale. Herein, an efficient L-theanine-producing strain of Corynebacterium glutamicum was constructed by combining protein and metabolic engineering. Firstly, a γ-glutamylmethylamide synthetase from Paracoccus aminovorans (PaGMAS) was isolated and engineered by computer-aided design, the resulting mutant E179K/N105R improved L-theanine yield by 36.61 %. Subsequently, to increase carbon flux towards L-theanine production, the gene ggt which degrades L-theanine, the gene alaT which participated in L-alanine synthesis, and the gene NCgl1221 which encodes glutamate-exporting protein were deleted. Finally, ppk gene was overexpressed to enhance intracellular ATP production. The reprogramed strain produced 44.12 g/L L-theanine with a yield of 57.11 % and productivity of 1.16 g/L/h, which is the highest L-theanine titer reported by Corynebacterium glutamicum. This study provides an efficient and economical biosynthetic pathway for the industrial production of L-theanine.
Topics: Corynebacterium glutamicum; Metabolic Engineering; Fermentation; Glutamic Acid; Glutamates
PubMed: 38103752
DOI: 10.1016/j.biortech.2023.130200 -
Scientific Reports Mar 2024In the Anthropocene, plastic pollution has become a new environmental biotope, the so-called plastisphere. In the oceans, nano- and micro-sized plastics are omnipresent...
In the Anthropocene, plastic pollution has become a new environmental biotope, the so-called plastisphere. In the oceans, nano- and micro-sized plastics are omnipresent and found in huge quantities throughout the water column and sediment, and their large surface area-to-volume ratio offers an excellent surface to which hydrophobic chemical pollutants (e.g. petrochemicals and POPs) can readily sorb to. Our understanding of the microbial communities that breakdown plastic-sorbed chemical pollutants, however, remains poor. Here, we investigated the formation of 500 nm and 1000 nm polystyrene (PS) agglomerations in natural seawater from a coastal environment, and we applied DNA-based stable isotope probing (DNA-SIP) with the 500 nm PS sorbed with isotopically-labelled phenanthrene to identify the bacterial members in the seawater community capable of degrading the hydrocarbon. Whilst we observed no significant impact of nanoplastic size on the microbial communities associated with agglomerates that formed in these experiments, these communities were, however, significantly different to those in the surrounding seawater. By DNA-SIP, we identified Arcobacteraceae, Brevundimonas, Comamonas, uncultured Comamonadaceae, Delftia, Sphingomonas and Staphylococcus, as well as the first member of the genera Acidiphilum and Pelomonas to degrade phenanthrene, and of the genera Aquabacterium, Paracoccus and Polymorphobacter to degrade a hydrocarbon. This work provides new information that feeds into our growing understanding on the fate of co-pollutants associated with nano- and microplastics in the ocean.
Topics: Microplastics; Plastics; Polystyrenes; Comamonadaceae; DNA Probes; Environmental Pollutants; Isotopes; Microbiota; Phenanthrenes; DNA
PubMed: 38433255
DOI: 10.1038/s41598-024-55825-9 -
Water Research Aug 2024Overuse of antibiotics has led to their existence in nitrogen-containing water. The impacts of antibiotics on bio-denitrification and the metabolic response of...
Overuse of antibiotics has led to their existence in nitrogen-containing water. The impacts of antibiotics on bio-denitrification and the metabolic response of denitrifiers to antibiotics are unclear. We systematically analyzed the effect of ciprofloxacin (CIP) on bio-denitrification and found that 5 mg/L CIP greatly inhibited denitrification with a model denitrifier (Paracoccus denitrificans). Nitrate reduction decreased by 32.89 % and nitrous oxide emission increased by 75.53 %. The balance analysis of carbon and nitrogen metabolism during denitrification showed that CIP exposure blocked electron transfer and reduced the flow of substrate metabolism used for denitrification. Proteomics results showed that CIP exposure induced denitrifiers to use the pentose phosphate pathway more for substrate metabolism. This caused a substrate preference to generate NADPH to prevent cellular damage rather than NADH for denitrification. Notably, despite denitrifiers having antioxidant defenses, they could not completely prevent oxidative damage caused by CIP exposure. The effect of CIP exposure on denitrifiers after removal of extracellular polymeric substances (EPS) demonstrated that EPS around denitrifiers formed a barrier against CIP. Fluorescence and infrared spectroscopy revealed that the binding effect of proteins in EPS to CIP prevented damage. This study shows that denitrifiers resist antibiotic stress through different intracellular and extracellular defense strategies.
Topics: Ciprofloxacin; Denitrification; Anti-Bacterial Agents; Paracoccus denitrificans
PubMed: 38851110
DOI: 10.1016/j.watres.2024.121851 -
Microbial Cell Factories Feb 2024To contribute to the discovery of new microbial strains with metabolic and physiological robustness and develop them into successful chasses, Paracoccus pantotrophus DSM...
BACKGROUND
To contribute to the discovery of new microbial strains with metabolic and physiological robustness and develop them into successful chasses, Paracoccus pantotrophus DSM 2944, a Gram-negative bacterium from the phylum Alphaproteobacteria and the family Rhodobacteraceae, was chosen. The strain possesses an innate ability to tolerate high salt concentrations. It utilizes diverse substrates, including cheap and renewable feedstocks, such as C1 and C2 compounds. Also, it can consume short-chain alkanes, predominately found in hydrocarbon-rich environments, making it a potential bioremediation agent. The demonstrated metabolic versatility, coupled with the synthesis of the biodegradable polymer polyhydroxyalkanoate, positions this microbial strain as a noteworthy candidate for advancing the principles of a circular bioeconomy.
RESULTS
The study aims to follow the chassis roadmap, as depicted by Calero and Nikel, and de Lorenzo, to transform wild-type P. pantotrophus DSM 2944 into a proficient SynBio (Synthetic Biology) chassis. The initial findings highlight the antibiotic resistance profile of this prospective SynBio chassis. Subsequently, the best origin of replication (ori) was identified as RK2. In contrast, the non-replicative ori R6K was selected for the development of a suicide plasmid necessary for genome integration or gene deletion. Moreover, when assessing the most effective method for gene transfer, it was observed that conjugation had superior efficiency compared to electroporation, while transformation by heat shock was ineffective. Robust host fitness was demonstrated by stable plasmid maintenance, while standardized gene expression using an array of synthetic promoters could be shown. pEMG-based scarless gene deletion was successfully adapted, allowing gene deletion and integration. The successful integration of a gene cassette for terephthalic acid degradation is showcased. The resulting strain can grow on both monomers of polyethylene terephthalate (PET), with an increased growth rate achieved through adaptive laboratory evolution.
CONCLUSION
The chassis roadmap for the development of P. pantotrophus DSM 2944 into a proficient SynBio chassis was implemented. The presented genetic toolkit allows genome editing and therewith the possibility to exploit Paracoccus for a myriad of applications.
Topics: Humans; Paracoccus pantotrophus; Prospective Studies; Plasmids; Paracoccus; Biodegradation, Environmental
PubMed: 38360576
DOI: 10.1186/s12934-024-02325-0 -
The Science of the Total Environment Jun 2024Benzo(a)pyrene (BaP) is posing serious threats to soil ecosystems and its bioremediation usually limited by environmental factors and microbial activity. Humic acid...
Benzo(a)pyrene (BaP) is posing serious threats to soil ecosystems and its bioremediation usually limited by environmental factors and microbial activity. Humic acid (HA), a ubiquitous heterogeneous organic matter, which could affect the fate of environmental pollutants. However, the impact of HA on bioremediation of organic contamination remains controversial. In the present study, the biodegradation of BaP by Paracoccus aminovorans HPD-2 with and without HA was explored. Approximately 87.4 % of BaP was biodegraded in the HPD-2 treatment after 5 days of incubation, whereas the addition of HA dramatically reduced BaP biodegradation to 56.0 %. The limited BaP biodegradation in the HA + HPD-2 treatment was probably due to the decrease of BaP bioavailability which induced by the adsorption of HA with unspecific interactions. The excitation-emission matrix (EEM) of fluorescence characteristics showed that strain HPD-2 was responsible for the presence of protein-like substances and the microbial original humic substances in the HPD-2 treatment. Addition of HA would result in the increase of soluble microbial humic-like material, which should ascribe to the biodegradation of BaP and probably utilization of HA. Furthermore, both the growth and survival of strain HPD-2 were inhibited in the HA + HPD-2 treatment, because of the limited available carbon source (i.e. BaP) at the presence of HA. The expression of gene1789 and gene2589 dramatically decreased in the HA + HPD-2 treatment, and this should be responsible for the decrease of BaP biodegradation as well. This study reveals the mechanism that HA affect the BaP biodegradation, and the decrease of biodegradation should ascribe to the interaction of HA and bacterial strain. Thus, the bioremediation strategies of PAHs need to consider the effects of organic matter in environment.
Topics: Humic Substances; Biodegradation, Environmental; Benzo(a)pyrene; Paracoccus; Soil Pollutants; Soil Microbiology
PubMed: 38537831
DOI: 10.1016/j.scitotenv.2024.171966 -
Bioresource Technology Sep 2023N,N-dimethylformamide is a toxic chemical solvent, which widely exists in industrial wastewater. Nevertheless, the relevant methods merely achieved non-hazardous...
N,N-dimethylformamide is a toxic chemical solvent, which widely exists in industrial wastewater. Nevertheless, the relevant methods merely achieved non-hazardous treatment of N,N-dimethylformamide. In this study, one efficient N,N-dimethylformamide degrading strain was isolated and developed for pollutant removal coupling with poly(3-hydroxybutyrate) (PHB) accumulation. The functional host was characterized as Paracoccus sp. PXZ, which could consume N,N-dimethylformamide as the nutrient substrate for cell reproduction. Whole-genome sequencing analysis confirmed that PXZ simultaneously possesses the essential genes for poly(3-hydroxybutyrate) synthesis. Subsequently, the approaches of nutrient supplementation and various physicochemical variables to strengthen poly(3-hydroxybutyrate) production were investigated. The optimal biopolymer concentration was 2.74 g·L with a poly(3-hydroxybutyrate) proportion of 61%, showing a yield of 0.29 g-PHB·g-fructose. Furthermore, N,N-dimethylformamide served as the special nitrogen matter that could realize a similar poly(3-hydroxybutyrate) accumulation. This study provided a fermentation technology coupling with N,N-dimethylformamide degradation, offering a new strategy for resource utilization of specific pollutants and wastewater treatment.
Topics: 3-Hydroxybutyric Acid; Dimethylformamide; Paracoccus; Environmental Pollutants; Polyesters; Hydroxybutyrates
PubMed: 37315624
DOI: 10.1016/j.biortech.2023.129318 -
Bioresource Technology Feb 2024Biochar has demonstrated the potential in mitigating NO emissions during composting. However, little is known about how microbial communities on biochar particles...
Relationships between different types of biochar and NO emissions during composting based on roles of nosZ-carrying denitrifying bacterial communities enriched on compost and biochar particles.
Biochar has demonstrated the potential in mitigating NO emissions during composting. However, little is known about how microbial communities on biochar particles interact with NO emissions. This study selected three types of biochar (corn stalk biochar (CSB), rape straw biochar (RSB), and bamboo charcoal (BC)) to investigate the relationship between NO emissions and denitrifying bacterial communities on compost and biochar particles. The results showed that NO emissions rate were higher in the thermophilic phase, and the average emissions rate of BC treatment were lower 40% and 26% than CSB and RSB, respectively. The nosZ-carrying denitrifying bacterial community played a key role in reducing NO emissions, and the network indicated that Rhizobium and Paracoccus on compost particles may have played major roles in reducing NO emissions, but only Paracoccus on biochar particles. Notably, BC enhanced the efficiency of NO emission reduction by enhancing the abundance of these key genera.
Topics: Denitrification; Charcoal; Composting; Nitrous Oxide; Soil; Bacteria; Soil Microbiology
PubMed: 38122996
DOI: 10.1016/j.biortech.2023.130214 -
The Science of the Total Environment Jun 2024Until now, bacteria able to degrade, 3,3'-iminodipropionitrile (IDPN), a neurotoxin that destroys vestibular hair cells, causing ototoxicity, culminating in irreversible...
Until now, bacteria able to degrade, 3,3'-iminodipropionitrile (IDPN), a neurotoxin that destroys vestibular hair cells, causing ototoxicity, culminating in irreversible movement disorders, had never been isolated. The aim of this study was to isolate a novel IDPN-biodegrading microorganism and characterize its metabolic pathway. Enrichment was performed by inoculating activated sludge from a wastewater treatment bioreactor that treated IDPN-contaminated wastewater in M9 salt medium, with IDPN as the sole carbon source. A bacterial strain with a spherical morphology that could grow at high concentrations was isolated on a solid medium. Growth of the isolated strain followed the Monod kinetic model. Based on the 16S rRNA gene, the isolate was Paracoccus communis. Whole-genome sequencing revealed that the isolated P. communis possessed the expected full metabolic pathway for IDPN biodegradation. Transcriptome analyses confirmed the overexpression of the gene encoding hydantoinase/oxoprolinase during the exponential growth phase under IDPN-fed conditions, suggesting that the enzyme involved in cleaving the imine bond of IDPN may promote IDPN biodegradation. Additionally, the newly discovered P. communis isolate seems to metabolize IDPN through cleavage of the imine bond in IDPN via nitrilase, nitrile hydratase, and amidase reactions. Overall, this study lays the foundation for the application of IDPN-metabolizing bacteria in the remediation of IDPN-contaminated environments.
Topics: Nitriles; Bioreactors; Wastewater; Biodegradation, Environmental; Paracoccus; Waste Disposal, Fluid; Water Pollutants, Chemical; RNA, Ribosomal, 16S
PubMed: 38580115
DOI: 10.1016/j.scitotenv.2024.172099 -
Environmental Technology May 2024Petroleum hydrocarbon contamination is a serious concern across the globe. Here, the capability of native bacterial consortium enriched from sediment samples of Map Ta...
Petroleum hydrocarbon contamination is a serious concern across the globe. Here, the capability of native bacterial consortium enriched from sediment samples of Map Ta Phut Industrial Estate (MTPIE), Rayong, Thailand was described. The distribution of PAHs was assessed from the sediment samples collected from MTPIE by GC-FID and the toxic unit (TU) was calculated to assess the potential ecological risk to the surrounding biota. This study investigated the degradation potential and determined the PAH-degrading bacterial cultures by enriching collected sediments in PAHs mixtures (naphthalene, phenanthrene, and pyrene). The TPH degradation capacity of each bacterial consortium was validated in a soil microcosm using aged crude oil-contaminated soil. The MTPIE sediments were highly contaminated with PAHs (843.99-3904.39 ng g) and posed extremely high ecological risks to benthic biota (TU > 1). The consortium S5-P most significantly removed naphthalene (90.03%) and phenanthrene (88.14%) while the highest removal of pyrene was achieved by the S3-P consortium. Other consortia only partially degraded the PAHs. The dominant microbes in the consortia were determined using PCR-DGGE, it was found that the PAH degrading consortia were known PAH degraders such as and species and unknown PAH degraders such as species The consortium S5-P showed the highest degradation capacity, removing 74.99% of TPHs in the soil microcosm. Furthermore, the inoculation of PAH-biodegrading bacterial consortia significantly promoted the catechol-2,3-dioxygenase (C23O) and dehydrogenase (DHA) activities which directly correlated with the degradation efficiency of petroleum hydrocarbons ( < 0.05).
Topics: Thailand; Geologic Sediments; Polycyclic Aromatic Hydrocarbons; Phenanthrenes; Pyrenes; Naphthalenes; Bacteria; Risk Assessment; Petroleum; Soil; Biodegradation, Environmental; Soil Microbiology
PubMed: 36527266
DOI: 10.1080/09593330.2022.2157758