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1.
Ecotoxicol Environ Saf ; 231: 113216, 2022 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-35065503

RESUMO

Procambarus clarkii (crayfish) accumulates a high concentration of Arsenic (As) from the aquatic environment and causes considerable human health risks. In this study, Limosilactobacillus fermentum GR-3 strain was isolated from "Jiangshui" and applied for As(III) adsorption and antioxidant abilities. Strain GR-3 removed 50.67% of 50 mg/L As(III) and exhibited the high antioxidant potential of DPPH (1,1-Diphenyl-2-picrylhydrazyl) (87.63%) and hydroxyl radical (74.51%) scavenging rate in vitro. P. clarkii was feed with strain GR-3, the results showed that As(III) concentration reduced, and residual level in hepatopancreas was decreased by 36%, compared to As(III)-exposed group (control). Gut microbial sequencing showed that strain GR-3 restores gut microbiota dysbiosis caused by As(III) exposure. Further application in the field scale was performed and revealed a decrease in As(III) accumulation and increasing 50% aquaculture production of the total output. In summary, feed-additive probiotic is recommended as a novel strategy to minimize aquaculture foods toxicity and safe human health.


Assuntos
Arsênio , Limosilactobacillus fermentum , Animais , Arsênio/metabolismo , Arsênio/toxicidade , Astacoidea , Hepatopâncreas/metabolismo , Humanos , Estresse Oxidativo
2.
Ecotoxicology ; 25(1): 234-47, 2016 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-26589947

RESUMO

Strain LZ-C, isolated from a petrochemical wastewater discharge site, was found to be resistant to heavy metals and to degrade various aromatic compounds, including naphenol, naphthalene, 2-methylnaphthalene and toluene. Data obtained from 16S rRNA gene sequencing showed that this strain was closely related to Delftia lacustris. The 5,889,360 bp genome of strain LZ-C was assembled into 239 contigs and 197 scaffolds containing 5855 predicted open reading frames (ORFs). Among these predicted ORFs, 464 were different from the type strain of Delftia. The minimal inhibitory concentrations were 4 mM, 30 µM, 2 mM and 1 mM for Cr(VI), Hg(II), Cd(II) and Pb(II), respectively. Both genome sequencing and quantitative real-time PCR data revealed that genes related to Chr, Czc and Mer family genes play important roles in heavy metal resistance in strain LZ-C. In addition, the Na(+)/H(+) antiporter NhaA is important for adaptation to high salinity resistance (2.5 M NaCl). The complete pathways of benzene and benzoate degradation were identified through KEGG analysis. Interestingly, strain LZ-C also degrades naphthalene but lacks the key naphthalene degradation gene NahA. Thus, we propose that strain LZ-C exhibits a novel protein with a function similar to NahA. This study is the first to reveal the mechanisms of heavy metal resistance and salinity tolerance in D. lacustris and to identify a potential 2-methylnaphthalene degradation protein in this strain. Through whole-genome sequencing analysis, strain LZ-C might be a good candidate for the bioremediation of heavy metals and polycyclic aromatic hydrocarbons.


Assuntos
Delftia/genética , Delftia/metabolismo , Genoma Bacteriano , Metais Pesados/metabolismo , Hidrocarbonetos Policíclicos Aromáticos/metabolismo , Poluentes do Solo/metabolismo , China , Reação em Cadeia da Polimerase em Tempo Real , Análise de Sequência de DNA
3.
World J Microbiol Biotechnol ; 30(6): 1819-27, 2014 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-24458877

RESUMO

Streptomyces trypsin is one of the serine proteinases in Streptomyces griseus and acts as a key mediator during cell growth and differentiation. S. griseus trypsin (SGT) could be successfully expressed in Pichia pastoris by engineering the natural propeptide APNP. In this study, the recombinant Exmt with peptide YVEF and the wild-type SGT were comparatively investigated in detail. The recombinant Exmt showed significantly increased thermostability which t(½) value was 3.89-fold of that of the SGT at 40 °C. Moreover, the catalytic efficiency (referring to the specificity constant, k cat/K m) and pH tolerance of Exmt were also improved. In silico modeling analysis uncovered that introduction of the peptide YVEF resulted in a broadened substrate binding pocket and closer catalytic triad (His57, Asp¹°² and Ser¹95). The intramolecular Hydrogen bonds and the cation π-interactions were also dramatically increased. The results indicated that engineering of the N-terminus with artificial peptides might be an effective approach for optimizing the properties of the target enzymes.


Assuntos
Proteínas de Bactérias/química , Proteínas de Bactérias/genética , Peptídeos/genética , Engenharia de Proteínas , Streptomyces griseus/enzimologia , Tripsina/química , Tripsina/genética , Motivos de Aminoácidos , Proteínas de Bactérias/metabolismo , Biocatálise , Estabilidade Enzimática , Cinética , Peptídeos/metabolismo , Proteínas Recombinantes de Fusão/química , Proteínas Recombinantes de Fusão/genética , Proteínas Recombinantes de Fusão/metabolismo , Streptomyces griseus/química , Streptomyces griseus/genética , Tripsina/metabolismo
4.
Chemosphere ; 356: 141927, 2024 May.
Artigo em Inglês | MEDLINE | ID: mdl-38593954

RESUMO

Numerous animal studies have demonstrated the toxicity of hexavalent chromium [Cr(VI)] and the bioremediative effects of probiotics on the composition and functions of gut microbiota. Since the precise mechanisms of Cr(VI) detoxification and its interactions with human gut microbiota were unknown, a novel dual-chamber simulated intestinal (DCSI) system was developed to maintain both the stability of the simulated system and the composition of the gut microbiota. Probiotic GR-1 was found to regulate intestinal gut microbiota, thereby reducing the toxicity of Cr(VI) within the DCSI system. The results indicate that Cr(VI) levels were reduced from 2.260 ± 0.2438 µg/g to 1.7086 ± 0.1950 µg/g in the gut microbiota cell pellet, and Cr(VI) permeability decreased from 0.5521 ± 0.1132 µg/L to 0.3681 ± 0.0178 µg/L after 48 h in simulated gut fluid. Additionally, the removal rate of 1,1-Diphenyl-2-picrylhydrazyl (DPPH), reducibility (Vitamin C), and total antioxidant capacity (T-AOC) increased by 50.83%, 31.70%, and 27.56%, respectively, following probiotic treatment. The increase in antioxidant capacity correlated with total Cr removal (P < 0.05, r from -0.80 to 0.73). 16S rRNA sequencing analysis showed that gut microbiota composition was reshaped by the addition of probiotics, which regulated the recovery of the functional gut microbiota to normal levels, rather than restoring the entire gut microbiota composition for community function. Thus, this study not only demonstrates the feasibility and stability of culturing gut microbiota but also offers a new biotechnological approach to synthesizing functional communities with functional strains for environmental risk management.


Assuntos
Cromo , Microbioma Gastrointestinal , Pediococcus acidilactici , Probióticos , Cromo/toxicidade , Cromo/metabolismo , Microbioma Gastrointestinal/efeitos dos fármacos , Humanos , Biodegradação Ambiental
5.
Food Chem ; 440: 138270, 2024 May 15.
Artigo em Inglês | MEDLINE | ID: mdl-38150908

RESUMO

Limited application in protecting lung health is attributed to the low levels of active compounds in lily plant bulbs. This study focused on enhancing the active compounds by fermenting Lilium davidii (Lanzhou Lily) bulbs with Limosilactobacillus fermentum GR-3, isolated from Jiangshui. Lily fermented bulbs with strain GR-3 (LFB+GR-3) increased the bioavailability of hexadecanoic acid methyl ester, 22-tetrahydroxy-5alpha-cholestan-6-one-3-O-beta-d-allopyranoside, 22-O-(6-deoxy-Alpha-l-mannopyranosyl)-3-O-beta-d-glucopyranosyl-pregn-5-en-20-one, 1-O-trans-feruloylglycerol, and 3,4 dihydroxybenzoic acid. LFB+GR-3 fraction was employed to treat the mice model exposed to the carbon black nanoparticles (CBNPs). Immunohistochemical analysis revealed that the deposition of CBNPs and damages in lung tissues were limited in the LFB+GR-3 treatment group, while TNF-α, IL-10, and IL-6 were elevated by 6.9, 4.3, and 7 folds in the CBNP exposure group. In addition, Lactobacillus, Escherichia, Lactococcus, and Muribacter were dominant in the lung microbiota of LFB+GR-3 than the CBNP group. The use of probiotic fermented lily bulbs might be helpful in lung infection treatment.


Assuntos
Lilium , Probióticos , Animais , Camundongos , Lilium/química , Plantas , Raízes de Plantas/química , Pulmão
6.
Appl Environ Microbiol ; 79(12): 3851-5, 2013 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-23563937

RESUMO

Applying in silico simulations and in vitro experiments, the amino acid proline was proved to have a profound influence on Streptomyces griseus trypsinogen, and the hydrogen bond between H(57) and D(102) was found to be crucial for trypsin activity. By introducing an artificial propeptide, IVEF, the titer of trypsin was increased 6.71-fold.


Assuntos
Reatores Biológicos , Modelos Moleculares , Pichia/metabolismo , Precursores de Proteínas/farmacologia , Streptomyces griseus/enzimologia , Tripsina/biossíntese , Tripsinogênio/metabolismo , Simulação por Computador , Eletroforese em Gel de Poliacrilamida , Vetores Genéticos/genética , Ligação de Hidrogênio , Precursores de Proteínas/administração & dosagem , Streptomyces griseus/química , Tripsinogênio/química
7.
Sci Total Environ ; 904: 166773, 2023 Dec 15.
Artigo em Inglês | MEDLINE | ID: mdl-37689204

RESUMO

The gut microbes thrive by utilizing host energy and, in return, provide valuable benefits, akin to the symbiotic relationship. To study the mutualistic association between the gut microbiota and host, a range of gut microbe populations (85 %, 66 %, 45 % and 38 % at the normal level) with comparable structures were constructed in broiler model. The results revealed that reductions in gut microbial population led to decreased energy consumption, resulting in increased host weight (10.26 %, 30.88 %, 17.65 % and - 12.77 %, respectively). Fecal metabolome revealed that among 85 % and 66 % of the normal population level, the gut microbes downregulated the immune-associated pathways of tryptophan metabolism and catecholamine biosynthesis, while the level of fatty acid oxidation was upregulated at 45 %. In the host, the concentration of gut microbes contributed to regulate functions related to lipid biosynthesis (from glycerophosphoserines to glycerophosphoethanolamines (9.63 %, 12.20 %, 6.66 % and 47.75 %) and glycerophosphocholines (10.78 %, 36.51 %, 2.00 % and 87.11 %)) and inflammation responses (methionine and betaine metabolism). From 85 % to 45 % of gut microbes, broiler showed an inhibited immunity (thymus gland, spleen, SIgG and IgA) and increased low-level inflammation response (ALT and T-SOD). However, at 38 %, the immune indexes exhibited an increase (thymus gland, spleen, SIgG, and IgA increased by 8.67 %, 8.50 %, 20.87 %, and 29.43 %, respectively), indicating the host lipid accumulation and inflammation response were negatively correlated with the immune reaction. Collectively, the gut microbiota maintains a symbiotic relationship with the host through the secretion of beneficial substances to interact with the host.


Assuntos
Microbioma Gastrointestinal , Animais , Microbioma Gastrointestinal/fisiologia , Galinhas , Inflamação , Lipídeos , Imunoglobulina A
8.
Food Res Int ; 170: 112960, 2023 08.
Artigo em Inglês | MEDLINE | ID: mdl-37316006

RESUMO

Gluten accumulation damages the proximal small intestine and causes celiac disease (CeD) which has not been effectively treated except by using a gluten-free diet. In this study, strain Bacillus subtilis LZU-GM was isolated from Pakistani traditional fermented sourdough and could degrade 73.7% of gluten in 24 h in vitro. Strain LZU-GM was employed for practical application to investigate gluten degradation in mice models. The results showed that strain LZU-GM was colonized in mice and the survival rate was around 0.95 % (P < 0.0001). The gluten degradation was 3-fold higher in the small intestine of the strain LZU-GM treated mice group remaining 1511.96 ng/mL of gluten peptides than the untreated mice group (6500.38 ng/mL). Immunochemical analysis showed that gluten-treated mice established positive antigliadin antibodies (AGA) in serum (IgA, IgG, and anti-TG2 antibodies) as compared to the strain LZU-GM treatment group. Furthermore, the number of IFN-γ, TNF-α, IL-10, and COX-2 cells decrease in the lamina propria of the strain LZU-GM treatment group (P < 0.0001). Microbial community bar plot analysis showed that Lactobacillus, Dubosiella, and Enterococcus genera were restored and stabilized in the LZU-GM treatment group while Blautia and Ruminococcus were found lower. The oral gavage of probiotic strain LZU-GM might be useful for gluten metabolism in the intestine during digestion and would be a long-term dietary treatment for CeD management.


Assuntos
Doença Celíaca , Microbioma Gastrointestinal , Microbiota , Animais , Camundongos , Bacillus subtilis , Glutens , Aditivos Alimentares
9.
Environ Pollut ; 324: 121349, 2023 May 01.
Artigo em Inglês | MEDLINE | ID: mdl-36870597

RESUMO

Serum uric acid elevation has been found in long-term nickel (Ni) exposure occupational workers, but the mechanism is unclear. In this study, the relationship between Ni exposure and uric acid elevation was explored in a cohort of 109 participants composed of a Ni-exposed workers group and a control group. The results showed that Ni concentration (5.70 ± 3.21 µg/L) and uric acid level (355.95 ± 67.87 µmol/L) in the serum were increased in the exposure group with a significant positive correlation (r = 0.413, p < 0.0001). The composition of gut microbiota and metabolome revealed that the abundance of uric acid-lowering bacteria, such as Lactobacillus, Lachnospiraceae_Unclassfied and Blautia were reduced while pathogenic bacteria including Parabacteriadies and Escherichia-Shigella were enriched in Ni group, accompanied by impaired intestinal degradation of purines and upregulated biosynthesis of primary bile acids. Consistent with human results, the mice experiments showed that Ni treatment significantly promotes uric acid elevation and systemic inflammation. Lactobacillus and Blautia in gut microbiota were reduced and inflammation-related taxa Alistipes and Mycoplasma were enriched in the Ni treatment. In addition, LC-MS/MS metabolomic analysis indicated that purine nucleosides were accumulated in mice feces, which increased purine absorption and uric acid elevation in the serum. In summary, this study provides evidence that UA elevation was correlated with heavy metals exposure and highlighted the role of gut microbiota in intestinal purine catabolism and in the pathogenesis of heavy metal-induced hyperuricemia.


Assuntos
Microbioma Gastrointestinal , Humanos , Animais , Camundongos , Ácido Úrico , Níquel/toxicidade , Cromatografia Líquida , Espectrometria de Massas em Tandem , Inflamação
10.
J Ind Microbiol Biotechnol ; 39(11): 1651-62, 2012 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-22842958

RESUMO

In the present study, the genes encoding trypsinogen and active trypsin from Streptomyces griseus were both cloned and expressed in the methylotrophic yeast Pichia pastoris with the α-factor secretion signal under the control of the alcohol oxidase promoter. The mature trypsin was successfully accumulated extracellularly in soluble form with a maximum amidase activity of 6.6 U ml(-1) (batch cultivation with flask cultivation) or 14.4 U ml(-1) (fed-batch cultivation with a 3-l fermentor). In contrast, the recombinant trypsinogen formed inclusion bodies and no activity was detected. Replacement of the trypsin propeptide Ala-Pro-Asn-Pro confirmed that its physiological function was as a repressor of activity. More importantly, our results proved that the propeptide inhibited the activity of trypsinogen after its successful folding.


Assuntos
Reatores Biológicos , Pichia/genética , Pichia/metabolismo , Proteínas Recombinantes/metabolismo , Streptomyces griseus/metabolismo , Tripsina/metabolismo , Amidoidrolases/metabolismo , Esterases/metabolismo , Fermentação , Corpos de Inclusão/metabolismo , Dobramento de Proteína , Proteínas Recombinantes/biossíntese , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Reprodutibilidade dos Testes , Streptomyces griseus/genética , Tripsina/biossíntese , Tripsina/química , Tripsina/genética , Tripsinogênio/antagonistas & inibidores , Tripsinogênio/genética , Tripsinogênio/metabolismo
11.
Trends Microbiol ; 30(1): 69-78, 2022 01.
Artigo em Inglês | MEDLINE | ID: mdl-34215486

RESUMO

Biomethanation of carbohydrates (e.g., lignocellulosic biomass) and lipids (e.g., waste oils) has been well studied. However, investigations on the biomethanation of protein-rich biowastes (PRBs) and associated microbial communities have not been reported. This review summarizes the challenges in the metabolic process of anaerobic digestion of PRBs and the microbial instability associated with it. We discuss the diversity of bacterial and archaeal communities via metagenomics under PRB mono- and codigestion. A stable community structure with enhanced metabolic activity is a core factor in PRB biomethanation. The application of strategies such as codigestion of PRBs with carbon-rich biomass and microbial stimulation/augmentation would make PRB biomethanation more feasible.


Assuntos
Reatores Biológicos , Metano , Anaerobiose , Archaea/genética , Archaea/metabolismo , Biocombustíveis , Metano/metabolismo
12.
Carbohydr Polym ; 277: 118894, 2022 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-34893296

RESUMO

Chitin refers to a natural biopolymer, which is economically significant to next-generation biorefineries. In this study, a novel high-yield method with cell surface-display chitosanase (CHI-1) was built to produce chitooligosaccharides (COS) from shrimp chaff through the co-fermentation in the presence of Bacillus subtilis and Acetobacter sp. Under the optimized co-fermentation conditions (5 g/L yeast extracts, 10 g/L KH2PO4, 6% ethanol, 50 g/L glucose), the final deproteinization (DP) and demineralization (DM) efficiency and the chitin yield were achieved as 94, 92 and 18%, respectively. The engineered E. coli BL21-pET23b(+)-NICHI maintained 81% of the initial enzyme activity after 40 days at room temperature. The crude CHI-1 was inactivated after one-day interacting with prepared chitosan. Moreover, E. coli BL21-pET23b(+)-NICHI still maintained excellent hydrolysis ability in 7 days, and the COS yield reached 41%. Accordingly, the proposed method exhibited excellent stability and a high hydrolysis efficiency to produce COS with whole engineered cells.


Assuntos
Glicosídeo Hidrolases/metabolismo , Oligossacarídeos/biossíntese , Animais , Quitosana/química , Decápodes , Escherichia coli/enzimologia , Fermentação , Oligossacarídeos/química
13.
NPJ Biofilms Microbiomes ; 8(1): 63, 2022 08 16.
Artigo em Inglês | MEDLINE | ID: mdl-35974020

RESUMO

Exposure to heavy metals (HMs) is a threat to human health. Although probiotics can detoxify HMs in animals, their effectiveness and mechanism of action in humans have not been studied well. Therefore, we conducted this randomized, double-blind, controlled trial on 152 occupational workers from the metal industry, an at-risk human population, to explore the effectiveness of probiotic yogurt in reducing HM levels. Participants were randomly assigned to two groups: one consumed probiotic yogurt containing the HM-resistant strain Pediococcus acidilactici GR-1 and the other consumed conventional yogurt for 12 weeks. Analysis of metal contents in the blood revealed that the consumption of probiotic yogurt resulted in a higher and faster decrease in copper (34.45%) and nickel (38.34%) levels in the blood than the consumption of conventional yogurt (16.41% and 27.57%, respectively). Metagenomic and metabolomic studies identified a close correlation between gut microbiota (GM) and host metabolism. Significantly enriched members of Blautia and Bifidobacterium correlated positively with the antioxidant capacities of GM and host. Further murine experiments confirmed the essential role of GM and protective effect of GR-1 on the antioxidative role of the intestine against copper. Thus, the use of probiotic yogurt may be an effective and affordable approach for combating toxic metal exposure through the protection of indigenous GM in humans.ClinicalTrials.gov identifier: ChiCTR2100053222.


Assuntos
Microbioma Gastrointestinal , Metais Pesados , Pediococcus acidilactici , Probióticos , Animais , Cobre , Humanos , Metaboloma , Camundongos
14.
Environ Pollut ; 289: 117885, 2021 Nov 15.
Artigo em Inglês | MEDLINE | ID: mdl-34388552

RESUMO

Phytoremediation causes a large quantity of phytoremediation residues rich in heavy metals (HMs). This kind of plant residue can be used as a substrate for anaerobic digestion (AD) to reduce the content of HM-containing biomass, but high concentrations of HMs will inhibit the digestion efficiency and reduce the conversion efficiency of plant residues. Bioaugmentation may be an effective method to improve the degradation efficiency and methane yield of plant residues rich in HMs. In this study, a cellulose-degrading anaerobic bacteria Paracoccus sp. Termed strain LZ-G1 was isolated from cow dung, which can degrade cellulose and simultaneously adsorb Cd2+. The Cd2+ (10 mg/L)-adsorbtion efficiency and cellulose (463.12 g/kg)-degradation rate were 65.1 % and 60.59 %, respectively. In addition, using the strain LZ-G1 bioaugmented Cd2+-containing plant residues and cow manure mixed AD system, the system's biogas and methane production significantly increased (98.97 % and 142.03 %, respectively). During the AD process, the strain LZ-G1 was successfully colonized in the digestion system. Furthermore, the microbial community analysis revealed that LZ-G1 bioaugmentation alleviates the toxicity of free Cd2+ to the microbial community in the AD system, regulates and restores the archaea genus dominant in the methanogenesis stage, and restores the relative abundance of dominant bacteria associated with biomass hydrolysis. The restoration of the microbial community increased the biogas yield and methane production rate. Thus, bioaugmentation provides an easy and a feasible method for the actual on-site treatment of HM-rich phytoremediation residues.


Assuntos
Cádmio , Esterco , Anaerobiose , Animais , Bovinos , Celulose , Digestão , Feminino
15.
Environ Pollut ; 276: 116634, 2021 May 01.
Artigo em Inglês | MEDLINE | ID: mdl-33592445

RESUMO

Lead (Pb2+) is easy to accumulate in fish which become a major source of Pb2+ exposure to humans. In this study, a recombinant Escherichia coli strain expressing Pb2+-specific surface-binding protein anchored by the ice nucleation protein was introduced into grass carp (Ctenopharyngodon idellus) gut to investigate its protective effect against dietary Pb2+ exposure. Pb2+ mostly precipitated on the surface of the engineered strain through Pb2+-specific surface-binding protein, with a maximum adsorption efficiency of 73% and an adsorption capacity of 163 µmol/g dry cells. The Pb2+ concentration in engineered bacteria-fed grass carp was reduced significantly, and the residual level of Pb2+ in feces was increased by 76%, compared with the control group. Meantime, the engineered bacteria were able to mitigate the oxidative stress and histological alterations of intestines and dysbiosis of gut microbiota induced by Pb2+exposure. Thus, the engineered bacterium that can effectively reduce Pb2+ residue in grass carp might be a useful tool for decontamintion of lead in aquatic organisms.


Assuntos
Carpas , Microbioma Gastrointestinal , Adsorção , Animais , Escherichia coli , Humanos , Chumbo
16.
Environ Pollut ; 278: 116801, 2021 Jun 01.
Artigo em Inglês | MEDLINE | ID: mdl-33689949

RESUMO

Anaerobic digestion, a promising technology for waste utilization and bioenergy generation, is a suitable approach to convert the shrimp waste to biomethane, reducing its environmental impact. In this study, shrimp chaff (SC) was co-digested corn straw (CS), wheat straw (WS), and sugarcane bagasse (SB). In co-digestion, SC enhanced biomethane production of CS by 8.47-fold, followed by SC + WS (5.67-folds), and SC + SB (3.37-folds). SC addition to agricultural biomass digestion also promoted the volatile solids removal up to 85%. Microbial community analysis of SC and CS co-digestion presented the dominance of phylum Bacteroidetes, Firmicutes, Proteobacteria, and Euryarchaeota. Proteolytic bacteria were dominant (18.02%) during co-digestion of SC and CS, with Proteiniphilum as major bacterial genera (14%) that converts complex proteinaceous substrates to organic acids. Among the archaeal community, Methanosarcina responsible for conversion of acetate and hydrogen to biomethane, increased up to 70.77% in SC and CS digestion. Addition of SC to the digestion of agricultural wastes can significantly improve the biomethane production along with its effective management to reduce environmental risks.


Assuntos
Metano , Microbiota , Anaerobiose , Biocombustíveis , Biomassa , Reatores Biológicos , Digestão
17.
J Hazard Mater ; 406: 124440, 2021 03 15.
Artigo em Inglês | MEDLINE | ID: mdl-33302188

RESUMO

Sulfonamide antibiotics (SAs) are excreted into the ecosystem unchanged through feces and urine because of their low adsorption and degradation in the guts of humans and animals. In this study, a novel whole-cell biocatalyst with fungal laccase on the cell surface of Escherichia coli Nissle 1917 was developed to degrade sulfadiazine (SDZ). Engineered strain EcN-IL showed laccase enzyme activity of 2 ± 1 U/mg dry weight of cell and degraded 37 ± 1% of SDZ at temperature 40 °C and pH 5 within 3 h in vitro. Strain EcN-IL with 500 mg/kg of SDZ was employed as a food supplement to feed chicken broilers, which can reduce the residue of SDZ in broiler manure by 58 ± 2% and also reduced dysbiosis of the gut microbiota due to overuse of antibiotics. The genetically engineered EcN-IL has laid a foundation for degrading SDZ in broilers and their manure.


Assuntos
Microbioma Gastrointestinal , Sulfadiazina , Animais , Antibacterianos , Bioengenharia , Galinhas , Ecossistema , Microbioma Gastrointestinal/genética , Humanos , Lacase/genética , Esterco , Microbiologia do Solo
18.
Microb Biotechnol ; 14(2): 465-478, 2021 03.
Artigo em Inglês | MEDLINE | ID: mdl-32578381

RESUMO

Molecular analyses relying on RNA, as a direct way to unravel active microbes and their functional genes, have received increasing attention from environmental researchers recently. However, extracting sufficient and high-quality total microbial RNA from seriously heavy metal-contaminated soils is still a challenge. In this study, the guanidine thiocyanate-high EDTA (GTHE) method was established and optimized for recovering high quantity and quality of RNA from long-term heavy metal-contaminated soils. Due to the low microbial biomass in the soils, we combined multiple strong denaturants and intense mechanical lysis to break cells for increasing RNA yields. To minimize RNAase and heavy metals interference on RNA integrity, the concentrations of guanidine thiocyanate and EDTA were increased from 0.5 to 0.625 ml g-1 soil and 10 to 100 mM, respectively. This optimized GTHE method was applied to seven severely contaminated soils, and the RNA recovery efficiencies were 2.80 ~ 59.41 µg g-1 soil. The total microbial RNA of non-Cr(VI) (NT) and Cr(VI)-treated (CT) samples was utilized for molecular analyses. The result of qRT-PCR demonstrated that the expressions of two tested genes, chrA and yieF, were respectively upregulated 4.12- and 62.43-fold after Cr(VI) treatment. The total microbial RNA extracted from NT and CT samples, respectively, reached to 26.70 µg and 30.75 µg, which were much higher than the required amount (5 µg) for metatranscriptomic library construction. Besides, ratios of mRNA read were more than 86%, which indicated the high-quality libraries constructed for metatranscriptomic analysis. In summary, the GTHE method is useful to study microbes of contaminated habitats.


Assuntos
Metais Pesados , Poluentes do Solo , Ácido Edético , Guanidinas , Metais Pesados/análise , RNA , Solo , Poluentes do Solo/análise , Tiocianatos
19.
Sci Total Environ ; 742: 140435, 2020 Nov 10.
Artigo em Inglês | MEDLINE | ID: mdl-32623159

RESUMO

Microbial remediation is a promising method to treat Cr(VI) in industrial wastewater. The remediation efficiency and stress-resistance ability of Cr(VI) remediation genes in microbes are the limiting factors for their application in industrial wastewater treatment. To screen novel highly efficient Cr(VI) remediation genes, comparative metatranscriptomic and metagenomic analyses were performed on long-term Cr(VI)-contaminated riparian soil with/without additional Cr(VI) treatment. The most suitable Cr(VI) treatment time was determined to be 30 min according to the high quality RNA yield and fold changes in gene expression. Six novel genes, which had complete open reading frames (ORFs) in metagenomic libraries, were identified from unculturable microbes. In the phenotypic functional assay, all novel genes enhanced the Cr(VI) resistance/reduction ability of E. coli. In the industrial wastewater treatment, E-mcr and E-gsr presented at least 50% Cr(VI) removal efficiencies in the presence of 200-600 µM of Cr(VI), without a decrease in efficiency over 17 days. The stress resistance assay showed that gsr increased the growth rate of E. coli by at least 30% under different extreme conditions, and thus, gsr was identified as a general stress-response gene. In the Cr valence distribution assay, E-mcr presented ~40 µM higher extracellular Cr (III) compared to E-yieF. Additionally, transmission electron microscopy (TEM) of E-mcr showed bulk black agglomerates on the cell surface. Thus, mcr was identified as a membrane chromate reductase gene. This research provides a new idea for studying novel highly efficient contaminant remediation genes from unculturable microbes.


Assuntos
Proteínas de Escherichia coli , Águas Residuárias , Cromo , Escherichia coli , Metagenômica , Oxirredutases
20.
Commun Biol ; 3(1): 242, 2020 05 15.
Artigo em Inglês | MEDLINE | ID: mdl-32415160

RESUMO

Heavy metal contamination in food endangers human health. Probiotics can protect animals and human against heavy metals, but the detoxification mechanism has not been fully clarified. Here, mice were supplemented with Pediococcus acidilactici strain BT36 isolated from Tibetan plateau yogurt, with strong antioxidant activity but no chromate reduction ability for 20 days to ensure gut colonization. Strain BT36 decreased chromate accumulation, reduced oxidative stress, and attenuated histological damage in the liver of mice. 16S rRNA and metatranscriptome sequencing analysis of fecal microbiota showed that BT36 reversed Cr(VI)-induced changes in gut microbial composition and metabolic activity. Specifically, BT36 recovered the expressions of 788 genes, including 34 inherent Cr remediation-relevant genes. Functional analysis of 10 unannotated genes regulated by BT36 suggested the existence of a new Cr(VI)-reduction gene in the gut microbiota. Thus, BT36 can modulate the gut microbiota in response to Cr(VI) induced oxidative stress and protect against Cr toxicity.


Assuntos
Cromatos/toxicidade , Microbioma Gastrointestinal/efeitos dos fármacos , Estresse Oxidativo , Pediococcus acidilactici/química , Probióticos/farmacologia , Iogurte/microbiologia , Ração Animal/análise , Animais , Dieta , Camundongos , Probióticos/administração & dosagem , Tibet
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