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Caproate production by microbial fermentation gained the advantages of sustainability and eco-friendliness, but challenged by sterile fermentation environment, necessity of organic electron donors. Here, a single-step electro-fermentation (EF) process of mixed culture was proposed for caprate production from rice straw. At the optimal potential of -0.8â¯V, caproate concentration, yield and selectivity in the neutral red (NR)-mediated EF system were 2.4â¯g/L, 0.2â¯g/g and 26.6â¯%. Long-term operation accumulated 5.3â¯g/L caproate with the yield and selectivity of 0.2â¯g/g and 34.2â¯% in the EF+NR system. Bioaugmentation by dosing chain-elongation microbial consortium further improved the caproate production, yield and selectivity to 9.1â¯g/L, 0.3â¯g/g and 41.5â¯%, respectively. The improved caproate production in the bioaugmented EF+NR system was likely due to the enhanced interspecies electron transfer, reconstructed microbial community, multiple electron donors and suitable pH environment. Present study offers a feasible strategy for cost-effective caprate production directly from waste biomass.
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Modifying cells with polymers on the surface can enable them to gain or enhance function with various applications, wherein the atom transfer radical polymerization (ATRP) has garnered significant potential due to its biocompatibility. However, specifically initiating ATRP from the cell surface for in-situ modification remains challenging. This study established a bacterial surface-initiated ATRP method and further applied it for enhanced Cr(VI) removal. The cell surface specificity was facilely achieved by cell surface labelling with azide substrates, following alkynyl ATRP initiator specifically anchoring with azide-alkyne click chemistry. Then, the ATRP polymerization was initiated from the cell surface, and different polymers were successfully applied to in-situ modification. Further analysis revealed that the modification of Shewanella oneidensis with poly (4-vinyl pyridine) and sodium polymethacrylate improved the heavy metal tolerance and enhanced the Cr(VI) removal rate of 2.6 times from 0.088 h-1 to 0.314 h-1. This work provided a novel idea for bacterial surface modification and would extend the application of ATRP in bioremediation.
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Nanocellulose (NC) is a promising biopolymer for various biomedical applications owing to its biocompatibility and low toxicity. However, it faces challenges in tissue engineering (TE) applications due to the inconsistency of the microenvironment within the NC-based scaffolds with target tissues, including anisotropy microstructure and biomechanics. To address this challenge, a facile swelling-induced nanofiber alignment and a novel in situ biomineralization reinforcement strategies were developed for the preparation of NC-based scaffolds with tunable anisotropic structure and mechanical strength for guiding the differentiation of bone marrow-derived mesenchymal stem cells for potential TE application. The bacterial cellulose (BC) and cellulose nanofibrils (CNFs) based scaffolds with tunable swelling anisotropic index in the range of 10-100 could be prepared by controlling the swelling medium. The in situ biomineralization efficiently reinforced the scaffolds with 2-4 times and 10-20 times modulus increasement for BC and CNFs, respectively. The scaffolds with higher mechanical strength were superior in supporting cell growth and proliferation, suggesting the potential application in TE application. This work demonstrated the feasibility of the proposed strategy in the preparation of scaffolds with mechanical anisotropy to induce cells-directed differentiation for TE applications.
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Biomineralización , Diferenciación Celular , Celulosa , Células Madre Mesenquimatosas , Nanofibras , Andamios del Tejido , Celulosa/química , Células Madre Mesenquimatosas/citología , Células Madre Mesenquimatosas/efectos de los fármacos , Andamios del Tejido/química , Diferenciación Celular/efectos de los fármacos , Anisotropía , Nanofibras/química , Ingeniería de Tejidos/métodos , Animales , Proliferación Celular/efectos de los fármacos , Células de la Médula Ósea/citologíaRESUMEN
Direct interspecies electron transfer (DIET) is essential for maintaining the function and stability of anaerobic microbial consortia. However, only limited natural DIET modes have been identified and DIET engineering remains highly challenging. In this study, an unnatural DIET between Shewanella oneidensis MR-1 (SO, electron donating partner) and Rhodopseudomonas palustris (RP, electron accepting partner) was artificially established by a facile living cell-cell click chemistry strategy. By introducing alkyne- or azide-modified monosaccharides onto the cell outer surface of the target species, precise covalent connections between different species in high proximity were realized through a fast click chemistry reaction. Remarkably, upon covalent connection, outer cell surface C-type cytochromes mediated DIET between SO and RP was achieved and identified, although this was never realized naturally. Moreover, this connection directly shifted the natural H2 mediated interspecies electron transfer (MIET) to DIET between SO and RP, which delivered superior interspecies electron exchange efficiency. Therefore, this work demonstrated a naturally unachievable DIET and an unprecedented MIET shift to DIET accomplished by cell-cell distance engineering, offering an efficient and versatile solution for DIET engineering, which extends our understanding of DIET and opens up new avenues for DIET exploration and applications.
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Química Clic , Rhodopseudomonas , Shewanella , Transporte de Electrón , Shewanella/metabolismo , Shewanella/química , Rhodopseudomonas/metabolismo , Rhodopseudomonas/química , Azidas/química , Azidas/metabolismo , Alquinos/químicaRESUMEN
In recent years, the rapid development of pig farming has led to a large quantity of heavy metal-polluted wastewater. Thus, it was desirable to develop a simple heavy metal detection method for fast monitoring of the wastewater from the pig farms. Therefore, there was an urgent need to develop a simple method for rapidly detecting heavy metal ions in pig farm wastewater. Herein, a simple electrochemical method for simultaneous detection of Cu2+ and Zn2+ was developed and applied to pig farm wastewater. With a glassy carbon electrode and anodic stripping voltammetry, simultaneous detection of Cu2+ and Zn2+ in water was achieved without the need for complicated electrode modification. Furthermore, it was found that the addition of Cd2+ can enhance the response current of the electrode to Zn2+, which increased the signal by eight times. After systematic optimization, the limit of detection (LOD) of 9.3 µg/L for Cu2+ and 45.3 µg/L for Zn2+ was obtained. Finally, it was successfully applied for the quantification of Cu2+ and Zn2+ with high accuracy in pig farm wastewater. This work provided a new and simple solution for fast monitoring of the wastewater from pig farms and demonstrated the potential of electrochemical measurement for application in modern animal husbandry.
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Cobre , Técnicas Electroquímicas , Granjas , Aguas Residuales , Zinc , Aguas Residuales/química , Aguas Residuales/análisis , Cobre/análisis , Cobre/química , Zinc/análisis , Zinc/química , Animales , Porcinos , Técnicas Electroquímicas/métodos , Contaminantes Químicos del Agua/análisis , Electrodos , Límite de DetecciónRESUMEN
Microbial polysaccharides (MPs) are biopolymers secreted by microorganisms such as bacteria and fungi during their metabolic processes. Compared to polysaccharides derived from plants and animals, MPs have advantages such as wide sources, high production efficiency, and less susceptibility to natural environmental influences. The most attractive feature of MPs lies in their diverse biological activities, such as antioxidative, anti-tumor, antibacterial, and immunomodulatory activities, which have demonstrated immense potential for applications in functional foods, cosmetics, and biomedicine. These bioactivities are precisely regulated by their sophisticated molecular structure. However, the mechanisms underlying this precise regulation are not yet fully understood and continue to evolve. This article presents a comprehensive review of the most representative species of MPs, including their fermentation and purification processes and their biomedical applications in recent years. In particular, this work presents an in-depth analysis into the structure-activity relationships of MPs across multiple molecular levels. Additionally, this review discusses the challenges and prospects of investigating the structure-activity relationships, providing valuable insights into the broad and high-value utilization of MPs.
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Antibacterianos , Antioxidantes , Animales , Antibacterianos/farmacología , Antioxidantes/farmacología , Transporte Biológico , Fermentación , Alimentos FuncionalesRESUMEN
Amyloid fibrils derived from different proteins have been proved as a promising material for adsorption of various pollutants from wastewater, which showed advantages of low cost and eco-friendliness. However, most of the amyloid fibrils derived from animal-based proteins with high environmental footprint, while more sustainable amyloid fibrils derived from plant materials are desirable. In this study, a plant-derived amyloid fibril was extracted from the commonly used wheat flour with a simple and scalable protein purification and fibrillization process. Interestingly, the amyloid fibrils showed good adsorption capacity towards typical organic dyes (Eosin Y (EY) and Congo red (CR)) from contaminated water. Adsorption kinetic analysis indicated the adsorption process to EY or CR by wheat flour amyloid well fitted with a pseudo-second-order model. The adsorption also followed a Langmuir isothermal model with adsorption capacities of 333 mg/g and 138 mg/g towards CR and EY, respectively. This work demonstrated the feasibility to utilize the plant-based amyloid fibril for organic dyes removal from contaminated water, which provided an affordable, sustainable and scalable tool for organic dyes removal from wastewater.
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Nanocellulose-based biomaterials have gained significant attention in various fields, especially in medical and pharmaceutical areas, due to their unique properties, including non-toxicity, high specific surface area, biodegradability, biocompatibility, and abundant feasible and sophisticated strategies for functional modification. The biosafety of nanocellulose itself is a prerequisite to ensure the safe and effective application of biomaterials as they interact with living cells, tissues, and organs at the nanoscale. Potential residual endogenous impurities and exogenous contaminants could lead to the failure of the intended functionalities or even serious health complications if they are not adequately removed and assessed before use. This review summarizes the sources of impurities in nanocellulose that may pose potential hazards to their biosafety, including endogenous impurities that co-exist in the cellulosic raw materials themselves and exogenous contaminants caused by external exposure. Strategies to reduce or completely remove these impurities are outlined and classified as chemical, physical, biological, and combined methods. Additionally, key points that require careful consideration in the interpretation of the biosafety evaluation outcomes were discussed to ensure the safety and effectiveness of the nanocellulose-based biomaterials in medical applications.
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Materiales Biocompatibles , Contención de Riesgos BiológicosRESUMEN
ß-mannanases are pivotal enzymes that cleave the mannan backbone to release short chain mannooligosaccharides, which have tremendous biotechnological applications including food/feed, prebiotics and biofuel production. Due to the high temperature conditions in many industrial applications, thermophilic mannanases seem to have great potential to overcome the thermal impediments. Thus, structural analysis of thermostable ß-mannanases is extremely important, as it could open up new avenues for genetic engineering, and protein engineering of these enzymes with enhanced properties and catalytic efficiencies. Under this scope, the present review provides a state-of-the-art discussion on the thermophilic ß-mannanases from bacterial origin, their production, engineering and structural characterization. It covers broad insights into various molecular biology techniques such as gene mutagenesis, heterologous gene expression, and protein engineering, that are employed to improve the catalytic efficiency and thermostability of bacterial mannanases for potential industrial applications. Further, the bottlenecks associated with mannanase production and process optimization are also discussed. Finally, future research related to bioengineering of mannanases with novel protein expression systems for commercial applications are also elaborated.
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Bacterias , beta-Manosidasa , beta-Manosidasa/química , Bacterias/metabolismo , Ingeniería Genética , Biotecnología/métodos , Mananos/química , BioingenieríaRESUMEN
Bio-photoelectrochemical cell (BPEC) is an emerging technology that can convert the solar energy into electricity or chemicals. However, traditional BPEC depending on abiotic electrodes is challenging for microbial/enzymatic catalysis because of the inefficient electron exchange. Here, electroactive bacteria (Shewanella loihica PV-4) were used to reduce graphene oxide (rGO) nanosheets and produce co-assembled rGO/Shewanella biohydrogel as a basic electrode. By adsorbing chlorophyll contained thylakoid membrane, this biohydrogel was fabricated as a photoanode that delivered maximum photocurrent 126 µA/cm3 under visible light. Impressively, the biohydrogel could be served as a cathode in BPEC by forming coculture system with genetically edited Clostridium ljungdahlii. Under illumination, the BPEC with above photoanode and cathode yielded â¼ 5.4 mM butyrate from CO2 reduction, 169 % increase compared to dark process. This work provided a new strategy (nanotechnology combined with synthetic biology) to achieve efficient bioelectricity and valuable chemical production in PBEC.
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Fuentes de Energía Bioeléctrica , Dióxido de Carbono , Grafito , Dióxido de Carbono/metabolismo , Butiratos , Hidrogeles , Electricidad , Luz , ElectrodosRESUMEN
Bioelectrochemical reactions using whole-cell biocatalysts are promising carbon-neutral approaches because of their easy operation, low cost, and sustainability. Bidirectional (outward or inward) electron transfer via exoelectrogens plays the main role in driving bioelectrochemical reactions. However, the low electron transfer efficiency seriously inhibits bioelectrochemical reaction kinetics. Here, a three dimensional and artificial nanoparticles-constituent inverse opal-indium tin oxide (IO-ITO) electrode is fabricated and employed to connect with exoelectrogens (Shewanella loihica PV-4). The above electrode collected 128-fold higher cell density and exhibited a maximum current output approaching 1.5 mA cm-2 within 24 h at anode mode. By changing the IO-ITO electrode to cathode mode, the exoelectrogens exhibited the attractive ability of extracellular electron uptake to reduce fumarate and 16 times higher reverse current than the commercial carbon electrode. Notably, Fe-containing oxide nanoparticles are biologically synthesized at both sides of the outer cell membrane and probably contributed to direct electron transfer with the transmembrane c-type cytochromes. Owing to the efficient electron exchange via artificial and biosynthetic nanoparticles, bioelectrochemical CO2 reduction is also realized at the cathode. This work not only explored the possibility of augmenting bidirectional electron transfer but also provided a new strategy to boost bioelectrochemical reactions by introducing biohybrid nanoparticles.
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Electrodos , Nanopartículas , Shewanella , Transporte de Electrón , Shewanella/metabolismo , Nanopartículas/química , Compuestos de Estaño/química , Técnicas Electroquímicas/métodos , Electroquímica , Dióxido de Carbono/metabolismo , Dióxido de Carbono/química , Fuentes de Energía BioeléctricaRESUMEN
Due to the limited resources and environmental problems associated with fossil fuels, there is a growing interest in utilizing renewable resources for the production of biofuels through microbial fermentation. Isobutanol is a promising biofuel that could potentially replace gasoline. However, its production efficiency is currently limited by the use of naturally isolated microorganisms. These naturally isolated microorganisms often encounter problems such as a limited range of substrates, low tolerance to solvents or inhibitors, feedback inhibition, and an imbalanced redox state. This makes it difficult to improve their production efficiency through traditional process optimization methods. Fortunately, recent advancements in genetic engineering technologies have made it possible to enhance microbial hosts for the increased production of isobutanol from renewable resources. This review provides a summary of the strategies and synthetic biology approaches that have been employed in the past few years to improve naturally isolated or non-natural microbial hosts for the enhanced production of isobutanol by utilizing different renewable resources. Furthermore, it also discusses the challenges that are faced by engineered microbial hosts and presents future perspectives to enhancing isobutanol production. KEY POINTS: ⢠Promising potential of isobutanol to replace gasoline ⢠Engineering of native and non-native microbial host for isobutanol production ⢠Challenges and opportunities for enhanced isobutanol production.
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Biocombustibles , Gasolina , Butanoles , Clonación MolecularRESUMEN
Pathogens cause various infectious diseases and high morbidity and mortality which is a global public health threat. The highly sensitive and specific detection is of significant importance for the effective treatment and intervention to minimise the impact. However, conventional detection methods including culture and molecular method gravely depend on expensive equipment and well-trained skilled personnel, limiting in the laboratory. It remains challenging to adapt in resource-limiting areas, e.g., low and middle-income countries (LMICs). To this end, low-cost, rapid, and sensitive detection tools with the capability of field testing e.g., a portable device for identification and quantification of pathogens, has attracted increasing attentions. Recently, paper-based microfluidic analytical devices (µPADs) have shown a promising tool for rapid and on-site diagnosis, providing a cost-effective and sensitive analytical approach for pathogens detection. The fast turn-round data collection may also contribute to better understanding of the risks and insights on mitigation method. In this paper, critical developments of µPADs for in-field detection of pathogens both for clinical diagnostics and environmental surveillance are reviewed. The future development, and challenges of µPADs for rapid and onsite detection of pathogens are discussed, including using the cross-disciplinary development with, emerging techniques such as deep learning and Internet of Things (IoT).
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Monitoreo del Ambiente , Microfluídica , Internet , Dispositivos Laboratorio en un Chip , LaboratoriosRESUMEN
Although dye-decolourising peroxidases (DyPs) are well-known for lignin degradation, a comprehensive understanding of their mechanism remains unclear. Therefore, studying the mechanism of lignin degradation by DyPs is necessary for industrial applications and enzyme engineering. In this study, a dye-decolourising peroxidase (CsDyP) gene from C. serinivorans was heterologously expressed and studied for its lignin degradation potential. Molecular docking analysis predicted the binding of 2, 2-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), veratryl alcohol (VA), 2, 6-dimethylphenol (2, 6- DMP), guaiacol (GUA), and lignin to the substrate-binding pocket of CsDyP. Evaluation of the enzymatic properties showed that CsDyP requires pH 4.0 and 30 °C for optimal activity and has a high affinity for ABTS. In addition, CsDyP is stable over a wide range of temperatures and pH and can tolerate 5.0 mM organic solvents. Low NaCl concentrations promoted CsDyP activity. Further, CsDyP significantly reduced the chemical oxygen demand decolourised alkali lignin (AL) and milled wood lignin (MWL). CsDyP targets the ß-O-4, CO, and CC bonds linking lignin's G, S, and H units to depolymerize and produce aromatic compounds. Overall, this study delivers valuable insights into the lignin degradation mechanism of CsDyP, which can benefit its industrial applications and lignin valorization.
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Lignina , Peroxidasa , Peroxidasa/metabolismo , Lignina/química , Simulación del Acoplamiento Molecular , Oxidación-Reducción , Peroxidasas/metabolismo , Colorantes/químicaRESUMEN
Butyrate production from renewable biomass shows great potential against climate change and over-consumption of fossil fuels. Herein, key operational parameters of a cathodic electro-fermentation (CEF) process were optimized for efficient butyrate production from rice straw by mixed culture. The cathode potential, controlled pH and initial substrate dosage were optimized at -1.0 V (vs Ag/AgCl), 7.0 and 30 g/L, respectively. Under the optimal conditions, 12.50 g/L butyrate with yield of 0.51 g/g-rice straw were obtained in batch-operated CEF system. In fed-batch mode, butyrate production significantly increased to 19.66 g/L with the yield of 0.33 g/g-rice straw, but 45.99% butyrate selectivity still needs to be improved in future. Enriched butyrate producing bacteria (Clostridium cluster XIVa and IV) with proportion of 58.75% on the 21st day of the fed-batch fermentation, contributed to the high-level butyrate production. The study provides a promising approach for efficient butyrate production from lignocellulosic biomass.
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Butiratos , Oryza , Fermentación , BiomasaRESUMEN
Whole-cell biosensors are an important class of analytical tools that offer the advantages of low cost, facile operation, and unique reproduction/regeneration ability. However, it has always been quite challenging to expand the sensing spectrum of the host. Here, a new approach to extend the cell sensing spectrum with biomineralized nanoparticles is developed. The nano-biohybrid design comprise biomineralized FeS nanoparticles firmly anchored onto the bacterium, Shewanella oneidensis MR-1, wherein the nanoparticles are wired to the cellular electron transfer machinery (MtrCAB/OmcA) of the bacterium, forming an artificial cyborgian redox machinery consisting of FeS-MtrCAB/OmcA-FccA. Strikingly, with this cyborgian redox machinery, the sensing spectrum of FeS hybridized S. oneidensis MR-1 cell is successfully expanded to enable whole-cell electrochemical detection of Vitamin B12, while an unhybridized native cell is incapable of sensing. This proof-of-concept nano-biohybrid design offers a new perspective on manipulating the microbial toolkit for an expanded sensing spectrum in whole-cell biosensors.
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Nanopartículas , Shewanella , Oxidación-Reducción , Transporte de ElectrónRESUMEN
A three-dimensional (3D) macroporous reduced graphene oxide/polypyrrole (rGO/Ppy) hydrogel assembled by bacterial cells was fabricated and applied for microbial fuel cells. By taking the advantage of electroactive cell-induced bioreduction of graphene oxide and in-situ polymerization of Ppy, a facile self-assembly by Shewanella oneidensis MR-1and in-situ polymerization approach for 3D rGO/Ppy hydrogel preparation was developed. This facile one-step self-assembly process enabled the embedding of living electroactive cells inside the hydrogel electrode, which showed an interconnected 3D macroporous structures with high conductivity and biocompatibility. Electrochemical analysis indicated that the self-assembly of cell-embedding rGO/Ppy hydrogel enhanced the electrochemical activity of the bioelectrode and reduced the electron charge transfer resistance between the cells and the electrode. Impressively, extremely high power output of 3366 ± 42 mW m-2 was achieved from the MFC with cell-embedding rGO/Ppy hydrogel rGO/Ppy, which was 8.6 times of that delivered from the MFC with bare electrode. Further analysis indicated that the increased cell loading by the hydrogel and improved electrochemical activity by the rGO/Ppy composite would be the underlying mechanism for this performance improvement. This study provided a facile approach to fabricate the biocompatible and electrochemical active 3D nanocomposites for MFC, which would also be promising for performance optimization of various bioelectrochemical systems.
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Fuentes de Energía Bioeléctrica , Fuentes de Energía Bioeléctrica/microbiología , Polímeros/química , Pirroles/química , Hidrogeles , ElectrodosRESUMEN
Butanol production by solventogenic Clostridia shows great potential to combat the energy crisis, but is still challenged by low butanol selectivity and high downstream cost. In this study, a novel cathodic electro-fermentation (CEF) system mediated by methyl viologen (MV) was proposed and sequentially optimized to obtain highly selective butanol production. Under the optimal conditions (-0.60 V cathode potential, 0.50 mM MV, 30 g/L glucose), 7.17 ± 0.55 g/L butanol production were achieved with the yield of 0.32 ± 0.02 g/g. With the supplement of 4 g/L butyric acid as co-substrate, butanol production further improved to 13.14 ± 1.14 g/L with butanol yield and selectivity as high as 0.43 ± 0.01 g/g and 90.44 ± 1.66%, respectively. The polarized electrode enabled the unbalanced fermentation towards butanol formation and MV further inhibited hydrogen production, both of which contributed to the high-level butanol production and selectivity. The MV-mediated CEF system is a promising approach for cost-effective bio-butanol production.
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Butanoles , Electrones , Fermentación , 1-Butanol , Ácido ButíricoRESUMEN
Soil microbial fuel cells (SMFCs) are an innovative device for soil-powered biosensors. However, the traditional SMFC sensors relied on anodic biosensing which might be unstable for long-term and continuous monitoring of toxic pollutants. Here, a carbon-felt-based cathodic SMFC biosensor was developed and applied for soil-powered long-term sensing of heavy metal ions. The SMFC-based biosensor generated output voltage about 400 mV with the external load of 1000 Ω. Upon the injection of metal ions, the voltage of the SMFC was increased sharply and quickly reached a stable output within 2~5 min. The metal ions of Cd2+, Zn2+, Pb2+, or Hg2+ ranging from 0.5 to 30 mg/L could be quantified by using this SMFC biosensor. As the anode was immersed in the deep soil, this SMFC-based biosensor was able to monitor efficiently for four months under repeated metal ions detection without significant decrease on the output voltage. This finding demonstrated the clear potential of the cathodic SMFC biosensor, which can be further implemented as a low-cost self-powered biosensor.
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Fuentes de Energía Bioeléctrica , Técnicas Biosensibles , Metales Pesados , Suelo , ElectrodosRESUMEN
A novel microfiber-like biohydrogel was fabricated by a facile approach relying on electroactive bacteria-induced graphene oxide reduction and confined self-assembly in a capillary tube. The microfiber-like biohydrogel (d = â¼1 mm) embedded high-density living cells and activated efficient electron exchange between cells and the conductive graphene network. Further, a miniature whole-cell electrochemical biosensing system was developed and applied for fumarate detection under -0.6 V (vs Ag/AgCl) applied potential. Taking advantage of its small size, high local cell density, and excellent electron exchange, this microfiber-like biohydrogel-based sensing system reached a linear calibration curve (R2 = 0.999) ranging from 1 nM to 10 mM. The limit of detection obtained was 0.60 nM, which was over 1300 times lower than a traditional biosensor for fumarate detection in 0.2 µL microdroplets. This work opened a new dimension for miniature whole-cell electrochemical sensing system design, which provided the possibility for bioelectrochemical detection in small volumes or three-dimensional local detection at high spatial resolutions.