RESUMO
In this study, highly selenite-resistant strains belonging to Brevundimonas diminuta (OK287021, OK287022) genus were isolated from previously operated single chamber microbial fuel cell (SCMFC). The central composite design showed that the B. diminuta consortium could reduce selenite. Under optimum conditions, 15.38 Log CFU mL-1 microbial growth, 99.08% Se(IV) reduction, and 89.94% chemical oxygen demand (COD) removal were observed. Moreover, the UV-visible spectroscopy (UV) and Fourier transform infrared spectroscopy (FTIR) analyses confirmed the synthesis of elemental selenium nanoparticles (SeNPs). In addition, transmission electron microscopy (TEM) and scanning electron microscope (SEM) revealed the formation of SeNPs nano-spheres. Besides, the bioelectrochemical performance of B. diminuta in the SCMFC illustrated that the maximum power density was higher in the case of selenite SCMFCs than those of the sterile control SCMFCs. Additionally, the bioelectrochemical impedance spectroscopy and cyclic voltammetry characterization illustrated the production of definite extracellular redox mediators that might be involved in the electron transfer progression during the reduction of selenite. In conclusion, B. diminuta whose electrochemical activity has never previously been reported could be a suitable and robust biocatalyst for selenite bioreduction along with wastewater treatment, bioelectricity generation, and economical synthesis of SeNPs in MFCs.
Assuntos
Fontes de Energia Bioelétrica , Oxirredução , Ácido Selenioso , Selênio , Selênio/metabolismo , Selênio/química , Ácido Selenioso/metabolismo , Caulobacteraceae/metabolismo , Nanopartículas/química , Eletricidade , Nanopartículas Metálicas/química , Consórcios Microbianos , Análise da Demanda Biológica de OxigênioRESUMO
BACKGROUND: Hydrocarbon pollution stemming from petrochemical activities is a significant global environmental concern. Bioremediation, employing microbial chitinase-based bioproducts to detoxify or remove contaminants, presents an intriguing solution for addressing hydrocarbon pollution. Chitooligosaccharides, a product of chitin degradation by chitinase enzymes, emerge as key components in this process. Utilizing chitinaceous wastes as a cost-effective substrate, microbial chitinase can be harnessed to produce Chitooligosaccharides. This investigation explores two strategies to enhance chitinase productivity, firstly, statistical optimization by the Plackett Burman design approach to evaluating the influence of individual physical and chemical parameters on chitinase production, Followed by response surface methodology (RSM) which delvs into the interactions among these factors to optimize chitinase production. Second, to further boost chitinase production, we employed heterologous expression of the chitinase-encoding gene in E. coli BL21(DE3) using a suitable vector. Enhancing chitinase activity not only boosts productivity but also augments the production of Chitooligosaccharides, which are found to be used as emulsifiers. RESULTS: In this study, we focused on optimizing the production of chitinase A from S. marcescens using the Plackett Burman design and response surface methods. This approach led to achieving a maximum activity of 78.65 U/mL. Subsequently, we cloned and expressed the gene responsible for chitinase A in E. coli BL21(DE3). The gene sequence, named SmChiA, spans 1692 base pairs, encoding 563 amino acids with a molecular weight of approximately 58 kDa. This sequence has been deposited in the NCBI GenBank under the accession number "OR643436". The purified recombinant chitinase exhibited a remarkable activity of 228.085 U/mL, with optimal conditions at a pH of 5.5 and a temperature of 65 °C. This activity was 2.9 times higher than that of the optimized enzyme. We then employed the recombinant chitinase A to effectively hydrolyze shrimp waste, yielding chitooligosaccharides (COS) at a rate of 33% of the substrate. The structure of the COS was confirmed through NMR and mass spectrometry analyses. Moreover, the COS demonstrated its utility by forming stable emulsions with various hydrocarbons. Its emulsification index remained stable across a wide range of salinity, pH, and temperature conditions. We further observed that the COS facilitated the recovery of motor oil, burned motor oil, and aniline from polluted sand. Gravimetric assessment of residual hydrocarbons showed a correlation with FTIR analyses, indicating the efficacy of COS in remediation efforts. CONCLUSIONS: The recombinant chitinase holds significant promise for the biological conversion of chitinaceous wastes into chitooligosaccharides (COS), which proved its potential in bioremediation efforts targeting hydrocarbon-contaminated sand.
Assuntos
Biodegradação Ambiental , Quitinases , Quitosana , Oligossacarídeos , Proteínas Recombinantes , Quitinases/metabolismo , Quitinases/genética , Oligossacarídeos/metabolismo , Animais , Quitosana/metabolismo , Proteínas Recombinantes/metabolismo , Proteínas Recombinantes/genética , Proteínas Recombinantes/biossíntese , Quitina/metabolismo , Hidrocarbonetos/metabolismo , Escherichia coli/metabolismo , Escherichia coli/genética , Crustáceos/metabolismo , Emulsificantes/metabolismo , Emulsificantes/químicaRESUMO
BACKGROUND: The application of exopolysaccharide-producing bacteria (EPS) in dual chamber microbial fuel cells (DCMFC) is critical which can minimize the chemical oxygen demand (COD) of molasses with bioelectricity production. Hence, our study aimed to evaluate the EPS production by the novel strain Bacillus piscis by using molasses waste. Therefore, statistical modeling was used to optimize the EPS production. Its structure was characterized by UV, FTIR, NMR, and monosaccharides compositions. Eventually, to highlight B. piscis' adaptability in energy applications, bioelectricity production by this organism was studied in the BCMFC fed by an optimized molasses medium. RESULTS: B. piscis OK324045 characterized by 16S rRNA is a potent EPS-forming organism and yielded a 6.42-fold increase upon supplementation of molasses (5%), MgSO4 (0.05%), and inoculum size (4%). The novel exopolysaccharide produced by Bacillus sp. (EPS-BP5M) was confirmed by the structural analysis. The findings indicated that the MFC's maximum close circuit voltage (CCV) was 265 mV. The strain enhanced the performance of DCMFC achieving maximum power density (PD) of 31.98 mW m-2, COD removal rate of 90.91%, and color removal of 27.68%. Furthermore, cyclic voltammetry (CV) revealed that anodic biofilms may directly transfer electrons to anodes without the use of external redox mediators. Additionally, CV measurements made at various sweep scan rates to evaluate the kinetic studies showed that the electron charge transfer was irreversible. The SEM images showed the biofilm growth distributed over the electrode's surface. CONCLUSIONS: This study offers a novel B. piscis strain for EPS-BP5M production, COD removal, decolorization, and electricity generation of the optimized molasses medium in MFCs. The biosynthesis of EPS-BP5M by a Bacillus piscis strain and its electrochemical activity has never been documented before. The approach adopted will provide significant benefits to sugar industries by generating bioelectricity using molasses as fuel and providing a viable way to improve molasses wastewater treatment.
Assuntos
Bacillus , Fontes de Energia Bioelétrica , Melaço , Cinética , RNA Ribossômico 16S , Eletricidade , EletrodosRESUMO
Microbial fuel cell (MFC) is used to remove organic pollutants while generating electricity. Biocathode plays as an efficient electrocatalyst for accelerating the Oxidation Reduction Reaction (ORR) of oxygen in MFC. This study integrated biocathode into a single-chamber microbial fuel cell (BSCMFC) to produce electricity from an organic substrate using aerobic activated sludge to gain more insights into anodic and cathodic biofilms. The maximum power density, current density, chemical oxygen demand (COD) removal, and coulombic efficiency were 0.593 W m-3, 2.6 A m-3, 83 ± 8.4%, and 22 ± 2.5%, respectively. Extracellular polymeric substances (EPS) produced by biofilm from the biocathode were higher than the bioanode. Infrared spectroscopy and Scanning Electron Microscope (SEM) examined confirmed the presence of biofilm by the adhesion on electrodes. The dominant phyla in bioanode were Proteobacteria, Firmicutes, Bacteroidetes, and Actinobacteria, while the dominant phylum in the biocathode was Proteobacteria. Therefore, this study demonstrates the applicable use of BSCMFC for bioelectricity generation and pollution control.
Assuntos
Fontes de Energia Bioelétrica , Biofilmes , Eletricidade , Eletrodos , Esgotos , Análise da Demanda Biológica de OxigênioRESUMO
Exopolysaccharides (EPSs) synthesized by lactic acid bacteria (LAB), have recently received much interest because of their various functional features in several industries. Food wastes (FWs) have become a major source of worry, as they can cause serious environmental contamination if improperly disposed. The utilization of these FWs is an excellent choice (approach) for producing value-added products such as EPSs, which will efficiently remediate wastes. The overall EPSs yield for the selected producers is strain-specific, and is heavily influenced by the nutritional and growing conditions used. This review emphasizes what is currently known about LAB's ability to generate economically relevant EPSs from FWs. In addition, a concise overview of the food industry, packaging, pharmaceutical and clinical applications application is discussed.
Assuntos
Indústria Alimentícia , Alimentos , Alimento Funcional , Lactobacillales/química , Polissacarídeos Bacterianos/análise , Eliminação de ResíduosRESUMO
Considering the need of new lactic acid bacteria (LAB) for the production of novel biosurfactant (BS) molecules, the current study brings out a new insight on the exploration of cheese samples for BS producers and process optimization for industrial applications. In view of this, Lactobacillus plantarum 60FHE, Lactobacillus paracasei 75FHE, and Lactobacillus paracasei 77FHE were selected as the most operative strains. The biosurfactants (BSs) described as glycolipoproteins via Fourier-transform infrared spectroscopy (FTIR) exhibited antimicrobial activity against the food-borne pathogens. L. plantarum 60FHE BS showed an anticancer activity against colon carcinoma cells and had a week antiviral activity against Hepatitis A virus. Furthermore, glycolipoprotein production was enhanced by 1.42-fold through the development of an optimized process using central composite design (CCD). Emulsifying activities were stable after 60-min incubation from 4 to 120 °C, at pH 2-12, and after the addition of NaCl (2-14%). Characterization by nuclear magnetic resonance spectroscopy (1H NMR) revealed that BS produced from strain 60FHE was glycolipoprotein. L. plantarum produced mixed BSs determined by Liquid Chromatography/Mass Spectrometry (LC-MS). Thus, indicating that BS was applied as a microbial food prevention and biomedical. Also, L. plantarum 60FHE BS was achieved with the use of statistical optimization on inexpensive food wastes.
Assuntos
Anti-Infecciosos/isolamento & purificação , Antineoplásicos/isolamento & purificação , Proteínas de Bactérias/isolamento & purificação , Queijo/microbiologia , Glicoproteínas/isolamento & purificação , Lactobacillus plantarum/química , Lipoproteínas/isolamento & purificação , Tensoativos/isolamento & purificação , Anti-Infecciosos/química , Anti-Infecciosos/economia , Anti-Infecciosos/farmacologia , Antineoplásicos/química , Antineoplásicos/economia , Antineoplásicos/farmacologia , Proteínas de Bactérias/química , Proteínas de Bactérias/economia , Proteínas de Bactérias/farmacologia , Linhagem Celular Tumoral , Cromatografia Líquida , Neoplasias do Colo/patologia , Glicoproteínas/química , Glicoproteínas/economia , Glicoproteínas/farmacologia , Bactérias Gram-Negativas/efeitos dos fármacos , Bactérias Gram-Positivas/efeitos dos fármacos , Hemólise/efeitos dos fármacos , Vírus da Hepatite A/efeitos dos fármacos , Humanos , Lacticaseibacillus paracasei/química , Lacticaseibacillus paracasei/isolamento & purificação , Lactobacillus plantarum/isolamento & purificação , Lipoproteínas/química , Lipoproteínas/economia , Lipoproteínas/farmacologia , Espectrometria de Massas , Ressonância Magnética Nuclear Biomolecular , Filogenia , Ribotipagem , Espectroscopia de Infravermelho com Transformada de Fourier , Tensoativos/química , Tensoativos/economia , Tensoativos/farmacologia , Resíduos/análiseRESUMO
Utilization of agricultural wastes as cheap natural resources for production of bioactive products is currently attracting global attention. For this purpose, this study focused on isolation of Aspergillus wewitschiae MN056175 as promising producer of inulinase, then investigating physiochemical, kinetics and thermodynamics of the obtained inulinase, and its ability to extract bioactive fructo-oligosaccharides (FOS) from Cynara scolymus leaves (artichoke leaves, AL). A. wewitschiae MN056175 inulinase gave the maximum activity at temperature 60 °C and inulin concentration 1%. The kinetics including Km and Vmax were determined to be 105.26 mg·ml-1 and 83.33 µmol·ml-1·min-1, respectively. The thermodynamics including, Ea (activation energy) and Ed (activation energy for denaturation) were determined to be 21.82 and 73.21 kJ·mol-1, Kd, T1/2, D-value, ΔH°, ΔG° and ΔS° at 40, 50 and 60 °C which indicated the stability of A. wewitschiae MN056175 inulinase. Moreover, this inulinase was capable of hydrolyzing Cynara scolymus leaves into reducing sugar and 15 FOS with different DP, total carbohydrate, and protein content under different conditions designed by central composite design (CCD). The 15 AL FOS showed different high antioxidant and prebiotic activities. Central FOS with probiotic bacteria exhibited significant antimicrobial activity against tested gram positive bacteria in a way higher than those recorded against gram negative bacteria.