RESUMO
The development and application of an electrochemical sensor is reported for detection of poly(3-hydroxybutyrate) (P3HB) - a bioplastic derived from agro-industrial residues. To overcome the challenges of molecular imprinting of macromolecules such as P3HB, this study employed methanolysis reaction to break down the P3HB biopolymer chains into methyl 3-hydroxybutyrate (M3HB) monomers. Thereafter, M3HB were employed as the target molecules in the construction of molecularly imprinted sensors. The electrochemical device was then prepared by electropolymerizing a molecularly imprinted poly (indole-3-acetic acid) thin film on a glassy carbon electrode surface modified with reduced graphene oxide (GCE/rGO-MIP) in the presence of M3HB. Electrochemical impedance spectroscopy (EIS), cyclic voltammetry (CV), scanning electron microscopy with field emission gun (SEM-FEG), Raman spectroscopy, attenuated total reflection Fourier-transform infrared (ATR-FTIR) and X-ray Photoelectron Spectroscopy (XPS) were employed to characterize the electrode surface. Under ideal conditions, the MIP sensor exhibited a wide linear working range of 0.1 - 10 nM and a detection limit of 0.3 pM (n = 3). The sensor showed good repeatability, selectivity, and stability over time. For the sensor application, the bioproduction of P3HB was carried out in a bioreactor containing the Burkholderia glumae MA13 strain and sugarcane byproducts as a supplementary carbon source. The analyses were validated through recovery assays, yielding recovery values between 102 and 104%. These results indicate that this MIP sensor can present advantages in the monitoring of P3HB during the bioconversion process.
Assuntos
Burkholderia , Técnicas Eletroquímicas , Eletrodos , Grafite , Hidroxibutiratos , Polímeros Molecularmente Impressos , Poliésteres , Grafite/química , Poliésteres/química , Hidroxibutiratos/química , Burkholderia/química , Burkholderia/metabolismo , Técnicas Eletroquímicas/métodos , Técnicas Eletroquímicas/instrumentação , Polímeros Molecularmente Impressos/química , Limite de Detecção , Oxirredução , Poli-HidroxibutiratosRESUMO
Poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) nanofibers embedded with borate glasses of 45B5 composition doped with Co2+, Cu2+, and Zn2 +(46.1 B2O326.9-X CaO24.4 Na2O2.6 P2O5, X CoO/CuO/ZnO mol % (X = 0-5)) were produced by electrospinning for wound healing applications. Prior to their addition, the glasses exhibited two broad halos typical of a vitreous borate network, which were mainly composed of ring-type metaborate structural units. The particle distribution in the PHBV nanofibers embedded with 45B5 borate bioactive glasses is present in isolated and agglomerated states, being partially coated by a polymeric layer-except for the cobalt-doped glass, which resulted in a successful encapsulation with 100% embedding efficiency. The incorporation of the glasses reduced the PHBV crystallinity degree and its decomposition temperature, as well as its mechanical properties, including Young's modulus, tensile strength, and elongation at break. The neat PHBV fibers and those containing the cobalt-doped glasses demonstrated great cytocompatibility with human keratinocytes (HaCat), as suggested by the high cell viability after 7 days of exposure. Further studies are needed to fully understand the wound healing potential of these fibers, but our results significantly contribute to the area.
Assuntos
Bandagens , Boratos , Cobalto , Cobre , Poliésteres , Zinco , Humanos , Cobre/química , Cobalto/química , Poliésteres/química , Boratos/química , Zinco/química , Vidro/química , Teste de Materiais , Cicatrização , Nanofibras/química , Linhagem Celular , Poli-HidroxibutiratosRESUMO
Dental implant success is threatened by peri-implantitis, an inflammation leading to implant failure. Conventional treatments struggle with the intricate microbial and host factors involved. Antibacterial membranes, acting as barriers and delivering antimicrobials, may offer a promising solution. Thus, this study highlights the potential of developing antibacterial membranes of poly-3-hydroxybutyrate and silver nanoparticles (Ag Nps) to address peri-implantitis challenges, discussing design and efficacy against potential pathogens. Electrospun membranes composed of PHB microfibers and Ag Nps were synthesized in a blend of DMF/chloroform at three different concentrations. Various studies were conducted on the characterization and antimicrobial activity of the membranes. The synthesized Ag Nps ranged from 4 to 8 nm in size. Furthermore, Young's modulus decreased, reducing from 13.308 MPa in PHB membranes without Ag Nps to 0.983 MPa in PHB membranes containing higher concentrations of Ag Nps. This demonstrates that adding Ag Nps results in a less stiff membrane. An increase in elongation at break was noted with the rise in Ag Nps concentration, from 23.597 % in PHB membranes to 60.136 % in PHB membranes loaded with Ag Nps. The antibiotic and antibiofilm activity of the membranes were evaluated against Pseudomonas aeruginosa, Staphylococcus aureus, Streptococcus mutans, and Candida albicans. The results indicated that all PHB membranes containing Ag Nps exhibited potent antibacterial activity by inhibiting the growth of biofilms and planktonic bacteria. However, inhibition of C. albicans occurred only with the PHB-Ag Nps C membrane. These findings emphasize the versatility and potential of Ag Nps-incorporated membranes as a multifunctional approach for preventing and addressing microbial infections associated with peri-implantitis. The combination of antibacterial and antibiofilm properties in these membranes holds promise for improving the management and treatment of peri-implantitis-related complications.
Assuntos
Antibacterianos , Biofilmes , Hidroxibutiratos , Membranas Artificiais , Nanopartículas Metálicas , Peri-Implantite , Prata , Prata/química , Prata/farmacologia , Biofilmes/efeitos dos fármacos , Antibacterianos/farmacologia , Antibacterianos/química , Nanopartículas Metálicas/química , Peri-Implantite/tratamento farmacológico , Peri-Implantite/microbiologia , Hidroxibutiratos/química , Hidroxibutiratos/farmacologia , Poliésteres/química , Testes de Sensibilidade Microbiana , Humanos , Staphylococcus aureus/efeitos dos fármacos , Pseudomonas aeruginosa/efeitos dos fármacos , Streptococcus mutans/efeitos dos fármacos , Poli-HidroxibutiratosRESUMO
Poly-hydroxybutyrate (PHB) is an environmentally friendly alternative for conventional fossil fuel-based plastics that is produced by various microorganisms. Large-scale PHB production is challenging due to the comparatively higher biomanufacturing costs. A PHB overproducer is the haloalkaliphilic bacterium Halomonas campaniensis, which has low nutritional requirements and can grow in cultures with high salt concentrations, rendering it resistant to contamination. Despite its virtues, the metabolic capabilities of H. campaniensis as well as the limitations hindering higher PHB production remain poorly studied. To address this limitation, we present HaloGEM, the first high-quality genome-scale metabolic network reconstruction, which encompasses 888 genes, 1528 reactions (1257 gene-associated), and 1274 metabolites. HaloGEM not only displays excellent agreement with previous growth data and experiments from this study, but it also revealed nitrogen as a limiting nutrient when growing aerobically under high salt concentrations using glucose as carbon source. Among different nitrogen source mixtures for optimal growth, HaloGEM predicted glutamate and arginine as a promising mixture producing increases of 54.2% and 153.4% in the biomass yield and PHB titer, respectively. Furthermore, the model was used to predict genetic interventions for increasing PHB yield, which were consistent with the rationale of previously reported strategies. Overall, the presented reconstruction advances our understanding of the metabolic capabilities of H. campaniensis for rationally engineering this next-generation industrial biotechnology platform. KEY POINTS: A comprehensive genome-scale metabolic reconstruction of H. campaniensis was developed. Experiments and simulations predict N limitation in minimal media under aerobiosis. In silico media design increased experimental biomass yield and PHB titer.
Assuntos
Halomonas , Hidroxibutiratos , Nitrogênio , Poliésteres , Poli-Hidroxibutiratos , Halomonas/metabolismo , Halomonas/genética , Halomonas/crescimento & desenvolvimento , Nitrogênio/metabolismo , Hidroxibutiratos/metabolismo , Poliésteres/metabolismo , Redes e Vias Metabólicas/genética , Biomassa , Glucose/metabolismoRESUMO
Inulin is a fructose-based polysaccharide that can be found in several plant species, from grass and onions to chicory roots; thus, it has the potential to be an excellent renewable source of fructose for several industrial applications. Among them, inulin hydrolysis can be coupled to a fermentation operation to produce polyhydroxybutyrate (PHB) using Cupriavidus necator H16. This work reports the PHB production process involving chicory root inulin hydrolysis using inulinase Novozym 960 followed by a C. necator fermentation. It was found that the maximum saccharification (95% wt.) was reached at 269 U/ginulin after 90 min. The hydrolysates obtained were then inoculated with C. necator, leading to a biomass concentration of 4 g/L with 30% (w/w) polymer accumulation. Although PHB production was low, during the first hours, the cell growth and polymer accumulation detected did not coincide with a fructose concentration decrease, suggesting a simultaneous saccharification and fermentation process, potentially alleviating the product inhibition inherent to the inulinase-fructose system. The characterization of the obtained PHB showed a polymer with more homogeneous values of Mw, and better thermal stability than PHB produced using pure fructose as a fermentation substrate. The results obtained demonstrate a viable alternative carbon substrate for PHB production, opening the possibility for inulin-rich renewable feedstock valorization.