Examinando por Autor "Valenzuela-Ibaceta, Felipe"
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Ítem Biosynthesis of Cu-In-S Nanoparticles by a Yeast Isolated from Union Glacier, Antarctica: A Platform for Enhanced Quantum Dot-Sensitized Solar Cells(Multidisciplinary Digital Publishing Institute (MDPI), 2024-03) Arriaza-Echanes, Carolina; Campo-Giraldo, Jessica L.; Valenzuela-Ibaceta, Felipe; Ramos-Zúñiga, Javiera; Pérez-Donoso, José M.In recent years, the utilization of extremophile microorganisms for the synthesis of metal nanoparticles, featuring enhanced properties and diverse compositions, has emerged as a sustainable strategy to generate high-quality nanomaterials with unique characteristics. Our study focuses on the biosynthesis of Cu-In-S (CIS) nanoparticles, which has garnered considerable attention in the past decade due to their low toxicity and versatile applications in biomedicine and solar cells. Despite this interest, there is a notable absence of reports on biological methods for CIS nanoparticle synthesis. In this research, three yeast species were isolated from soil samples in an extreme Antarctic environment—Union Glacier, Ellsworth Mountains. Among these isolates, Filobasidium stepposum demonstrated the capability to biosynthesize CIS nanoparticles when exposed to copper sulfate, indium chloride, glutathione, and cysteine. Subsequent purification and spectroscopic characterization confirmed the presence of characteristic absorbance and fluorescence peaks for CIS nanoparticles at 500 and 650 nm, respectively. Transmission electron microscopy analysis revealed the synthesis of monodisperse nanoparticles with a size range of 3–5 nm. Energy dispersive X-ray spectroscopy confirmed the composition of the nanoparticles, revealing the presence of copper, indium, and sulfur. The copper/indium ratio ranged from 0.15 to 0.27, depending on the reaction time. The biosynthesized CIS nanoparticles showed higher photostability than biomimetic nanoparticles and demonstrated successful application as photosensitizers in quantum dot-sensitized solar cells (QDSSC), achieving a conversion efficiency of up to 0.0247%. In summary, this work presents a cost-effective, straightforward, and environmentally friendly method for CIS nanoparticle synthesis. Furthermore, it constitutes the first documented instance of a biological procedure for producing these nanoparticles, opening avenues for the development of environmentally sustainable solar cells.Ítem Biosynthesis of photostable CdS quantum dots by UV-resistant psychrotolerant bacteria isolated from Union Glacier, Antarctica(BioMed Central Ltd, 2024-12) Vargas-Reyes, Matías; Bruna, Nicolás; Ramos-Zúñiga, Javiera; Valenzuela-Ibaceta, Felipe; Rivas-Álvarez, Paula; Navarro, Claudio A.; Pérez-Donoso, José MBackground: Quantum Dots (QDs) are fluorescent nanoparticles with exceptional optical and optoelectronic properties, finding widespread utility in diverse industrial applications. Presently, chemically synthesized QDs are employed in solar cells, bioimaging, and various technological domains. However, many applications demand QDs with prolonged lifespans under conditions of high-energy radiation. Over the past decade, microbial biosynthesis of nanomaterials has emerged as a sustainable and cost-effective process. In this context, the utilization of extremophile microorganisms for synthesizing QDs with unique properties has recently been reported. Results: In this study, UV-resistant bacteria were isolated from one of the most extreme environments in Antarctica, Union Glacier at the Ellsworth Mountains. Bacterial isolates, identified through 16 S sequencing, belong to the genera Rhodococcus, Pseudarthrobacter, and Arthrobacter. Notably, Rhodococcus sp. (EXRC-4 A-4), Pseudarthrobacter sp. (RC-2-3), and Arthrobacter sp. (EH-1B-1) tolerate UV-C radiation doses ≥ 120 J/m². Isolated UV-resistant bacteria biosynthesized CdS QDs with fluorescence intensities 4 to 8 times higher than those biosynthesized by E. coli, a mesophilic organism tolerating low doses of UV radiation. Transmission electron microscopy (TEM) analysis determined QD sizes ranging from 6 to 23 nm, and Fourier-transform infrared (FTIR) analysis demonstrated the presence of biomolecules. QDs produced by UV-resistant Antarctic bacteria exhibit high photostability after exposure to UV-B radiation, particularly in comparison to those biosynthesized by E. coli. Interestingly, red fluorescence-emitting QDs biosynthesized by Rhodococcus sp. (EXRC-4 A-4) and Arthrobacter sp. (EH-1B-1) increased their fluorescence emission after irradiation. Analysis of methylene blue degradation after exposure to irradiated QDs biosynthesized by UV-resistant bacteria, indicates that the QDs transfer their electrons to O2 for the formation of reactive oxygen species (ROS) at different levels. Conclusions: UV-resistant Antarctic bacteria represent a novel alternative for the sustainable generation of nanostructures with increased radiation tolerance—two characteristics favoring their potential application in technologies requiring continuous exposure to high-energy radiation. Graphical abstract: (Figure presented.) © The Author(s) 2024.Ítem Minicells as an Escherichia coli mechanism for the accumulation and disposal of fluorescent cadmium sulphide nanoparticles(BioMed Central Ltd, 2024-12) Valenzuela-Ibaceta, Felipe; Torres-Olea, Nicolás ; Ramos-Zúñiga, Javiera ; Dietz-Vargas, Claudio ; Navarro, Claudio A. ; Pérez-Donoso, José M.Background Bacterial biosynthesis of fluorescent nanoparticles or quantum dots (QDs) has emerged as a unique mechanism for heavy metal tolerance. However, the physiological pathways governing the removal of QDs from bacterial cells remains elusive. This study investigates the role of minicells, previously identified as a means of eliminating damaged proteins and enhancing bacterial resistance to stress. Building on our prior work, which unveiled the formation of minicells during cadmium QDs biosynthesis in Escherichia coli, we hypothesize that minicells serve as a mechanism for the accumulation and detoxification of QDs in bacterial cells. Results Intracellular biosynthesis of CdS QDs was performed in E. coli mutants Delta minC and Delta minCDE, known for their minicell-producing capabilities. Fluorescence microscopy analysis demonstrated that the generated minicells exhibited fluorescence emission, indicative of QD loading. Transmission electron microscopy (TEM) confirmed the presence of nanoparticles in minicells, while energy dispersive spectroscopy (EDS) revealed the coexistence of cadmium and sulfur. Cadmium quantification through flame atomic absorption spectrometry (FAAS) demonstrated that minicells accumulated a higher cadmium content compared to rod cells. Moreover, fluorescence intensity analysis suggested that minicells accumulated a greater quantity of fluorescent nanoparticles, underscoring their efficacy in QD removal. Biosynthesis dynamics in minicell-producing strains indicated that biosynthesized QDs maintained high fluorescence intensity even during prolonged biosynthesis times, suggesting continuous QD clearance in minicells. Conclusions These findings support a model wherein E. coli utilizes minicells for the accumulation and removal of nanoparticles, highlighting their physiological role in eliminating harmful elements and maintaining cellular fitness. Additionally, this biosynthesis system presents an opportunity for generating minicell-coated nanoparticles with enhanced biocompatibility for diverse applications.Ítem Production of minicell-like structures by Escherichia coli biosynthesizing cadmium fluorescent nanoparticles: a novel response to heavy metal exposure(BioMed Central Ltd, 0025) Valenzuela-Ibaceta, Felipe; Álvarez, Sergio A.; Pérez-Donoso, José M.The bacterial synthesis of fluorescent semiconductor nanoparticles or quantum dots (QDs), presents a sustainable method for producing nanomaterials with customized optical properties and significant technological potential. However, the underlying cellular mechanisms for this process remain elusive. Specifically, the role of cellular structures in QD generation has not been thoroughly investigated. In this study, we examined the morphological changes in Escherichia coli during the biosynthesis of cadmium sulfide (CdS) QDs, using a strain overexpressing the gshA gene to promote QD biosynthesis through increased glutathione (GSH) levels. Microscopy analyses revealed that fluorescence emission associated with QDs was concentrated at the cell poles, along with fluorescence emission from small spherical cells, a phenomenon exclusively detectable during QD biosynthesis. Transmission electron microscopy (TEM) revealed electron-dense nanomaterials localized at the cell poles. Furthermore, it was demonstrated the formation of minicell-like structures (∼ 0.5 μm in diameter) originating from these poles under biosynthesis conditions. These minicells encapsulated nanometric electron-dense material. Additional analyses indicated that minicells contained inclusion bodies, likely formed due to gshA overexpression and cadmium stress. Our findings confirms the role of minicells as a bacterial mechanism for sequestering cadmium at the cell poles and expelling the metal in the form of nanoparticles. This underscores the importance of minicells in bacterial physiology and stress responses, introducing a novel mechanism for heavy metal detoxification in bacteria. © The Author(s) 2025.Ítem Production of minicell-like structures by Escherichia coli biosynthesizing cadmium fluorescent nanoparticles: a novel response to heavy metal exposure(BioMed Central Ltd, 0025) Valenzuela-Ibaceta, Felipe; Álvarez, Sergio A.; Pérez-Donoso, José M.The bacterial synthesis of fluorescent semiconductor nanoparticles or quantum dots (QDs), presents a sustainable method for producing nanomaterials with customized optical properties and significant technological potential. However, the underlying cellular mechanisms for this process remain elusive. Specifically, the role of cellular structures in QD generation has not been thoroughly investigated. In this study, we examined the morphological changes in Escherichia coli during the biosynthesis of cadmium sulfide (CdS) QDs, using a strain overexpressing the gshA gene to promote QD biosynthesis through increased glutathione (GSH) levels. Microscopy analyses revealed that fluorescence emission associated with QDs was concentrated at the cell poles, along with fluorescence emission from small spherical cells, a phenomenon exclusively detectable during QD biosynthesis. Transmission electron microscopy (TEM) revealed electron-dense nanomaterials localized at the cell poles. Furthermore, it was demonstrated the formation of minicell-like structures (∼ 0.5 μm in diameter) originating from these poles under biosynthesis conditions. These minicells encapsulated nanometric electron-dense material. Additional analyses indicated that minicells contained inclusion bodies, likely formed due to gshA overexpression and cadmium stress. Our findings confirms the role of minicells as a bacterial mechanism for sequestering cadmium at the cell poles and expelling the metal in the form of nanoparticles. This underscores the importance of minicells in bacterial physiology and stress responses, introducing a novel mechanism for heavy metal detoxification in bacteria. © The Author(s) 2025.Ítem Production of minicell-like structures by Escherichia coli biosynthesizing cadmium fluorescent nanoparticles: a novel response to heavy metal exposure(BioMed Central Ltd, 2025-12) Valenzuela-Ibaceta, Felipe; Álvarez, Sergio A.; Pérez-Donoso, José M.The bacterial synthesis of fluorescent semiconductor nanoparticles or quantum dots (QDs), presents a sustainable method for producing nanomaterials with customized optical properties and significant technological potential. However, the underlying cellular mechanisms for this process remain elusive. Specifically, the role of cellular structures in QD generation has not been thoroughly investigated. In this study, we examined the morphological changes in Escherichia coli during the biosynthesis of cadmium sulfide (CdS) QDs, using a strain overexpressing the gshA gene to promote QD biosynthesis through increased glutathione (GSH) levels. Microscopy analyses revealed that fluorescence emission associated with QDs was concentrated at the cell poles, along with fluorescence emission from small spherical cells, a phenomenon exclusively detectable during QD biosynthesis. Transmission electron microscopy (TEM) revealed electron-dense nanomaterials localized at the cell poles. Furthermore, it was demonstrated the formation of minicell-like structures (∼ 0.5 μm in diameter) originating from these poles under biosynthesis conditions. These minicells encapsulated nanometric electron-dense material. Additional analyses indicated that minicells contained inclusion bodies, likely formed due to gshA overexpression and cadmium stress. Our findings confirms the role of minicells as a bacterial mechanism for sequestering cadmium at the cell poles and expelling the metal in the form of nanoparticles. This underscores the importance of minicells in bacterial physiology and stress responses, introducing a novel mechanism for heavy metal detoxification in bacteria.Ítem Production of minicell-like structures by Escherichia coli biosynthesizing cadmium fluorescent nanoparticles: a novel response to heavy metal exposure(BioMed Central Ltd, 0025-12) Valenzuela-Ibaceta, Felipe; Álvarez, Sergio A.; Pérez-Donoso, José M.The bacterial synthesis of fluorescent semiconductor nanoparticles or quantum dots (QDs), presents a sustainable method for producing nanomaterials with customized optical properties and significant technological potential. However, the underlying cellular mechanisms for this process remain elusive. Specifically, the role of cellular structures in QD generation has not been thoroughly investigated. In this study, we examined the morphological changes in Escherichia coli during the biosynthesis of cadmium sulfide (CdS) QDs, using a strain overexpressing the gshA gene to promote QD biosynthesis through increased glutathione (GSH) levels. Microscopy analyses revealed that fluorescence emission associated with QDs was concentrated at the cell poles, along with fluorescence emission from small spherical cells, a phenomenon exclusively detectable during QD biosynthesis. Transmission electron microscopy (TEM) revealed electron-dense nanomaterials localized at the cell poles. Furthermore, it was demonstrated the formation of minicell-like structures (∼ 0.5 μm in diameter) originating from these poles under biosynthesis conditions. These minicells encapsulated nanometric electron-dense material. Additional analyses indicated that minicells contained inclusion bodies, likely formed due to gshA overexpression and cadmium stress. Our findings confirms the role of minicells as a bacterial mechanism for sequestering cadmium at the cell poles and expelling the metal in the form of nanoparticles. This underscores the importance of minicells in bacterial physiology and stress responses, introducing a novel mechanism for heavy metal detoxification in bacteria. © The Author(s) 2025.Ítem Production of minicell-like structures by Escherichia coli biosynthesizing cadmium fluorescent nanoparticles: a novel response to heavy metal exposure(BioMed Central Ltd, 2025-12) Valenzuela-Ibaceta, Felipe; Álvarez, Sergio A.; Pérez-Donoso, José M.The bacterial synthesis of fluorescent semiconductor nanoparticles or quantum dots (QDs), presents a sustainable method for producing nanomaterials with customized optical properties and significant technological potential. However, the underlying cellular mechanisms for this process remain elusive. Specifically, the role of cellular structures in QD generation has not been thoroughly investigated. In this study, we examined the morphological changes in Escherichia coli during the biosynthesis of cadmium sulfide (CdS) QDs, using a strain overexpressing the gshA gene to promote QD biosynthesis through increased glutathione (GSH) levels. Microscopy analyses revealed that fluorescence emission associated with QDs was concentrated at the cell poles, along with fluorescence emission from small spherical cells, a phenomenon exclusively detectable during QD biosynthesis. Transmission electron microscopy (TEM) revealed electron-dense nanomaterials localized at the cell poles. Furthermore, it was demonstrated the formation of minicell-like structures (∼ 0.5 μm in diameter) originating from these poles under biosynthesis conditions. These minicells encapsulated nanometric electron-dense material. Additional analyses indicated that minicells contained inclusion bodies, likely formed due to gshA overexpression and cadmium stress. Our findings confirms the role of minicells as a bacterial mechanism for sequestering cadmium at the cell poles and expelling the metal in the form of nanoparticles. This underscores the importance of minicells in bacterial physiology and stress responses, introducing a novel mechanism for heavy metal detoxification in bacteria.Ítem Steady State Kinetics for Enzymes with Multiple Binding Sites Upstream of the Catalytic Site(MDPI, 2024-12-12) Osorio, Manuel I.; Petrache, Mircea; Salinas, Dino G.; Valenzuela-Ibaceta, Felipe; González-Nilo, Fernando; Tiznado, William; Pérez-Donoso, José M.; Bravo, Denisse; Yáñez, OsvaldoThe Michaelis–Menten mechanism, which describes the binding of a substrate to an enzyme, is a simplification of the process on a molecular scale. A more detailed model should include the binding of the substrate to precatalytic binding sites (PCBSs) prior to the transition to the catalytic site. Our work shows that the incorporation of PCBSs, in steady-state conditions, generates a Michaelis–Menten-type expression, in which the kinetic parameters KM and Vmax adopt more complex expressions than in the model without PCBSs. The equations governing reaction kinetics can be seen as generalized symmetries, relative to time translation actions over the state space of the underlying chemical system. The study of their structure and defining parameters can be interpreted as looking for invariants associated with these time evolution actions. The expression of (Formula presented.) decreases as the number of PCBSs increases, while (Formula presented.) reaches a minimum when the first PCBSs are incorporated into the model. To evaluate the trend of the dynamic behavior of the system, numerical simulations were performed based on schemes with different numbers of PCBSs and six conditions of kinetic constants. From these simulations, with equal kinetic constants for the formation of the Substrate/PCBS complex, it is observed that (Formula presented.) and (Formula presented.) are lower than those obtained with the Michaelis–Menten model. For the model with PCBSs, the (Formula presented.) reaches a minimum at one PCBS and that value is maintained for all of the systems evaluated. Since (Formula presented.) decreases with the number of PCBSs, the catalytic efficiency increases for enzymes fitting this model. All of these observations are consistent with the general equation obtained. This study allows us to explain, on the basis of the PCBS to (Formula presented.) and (Formula presented.) ratios, the effect on enzyme parameters due to mutations far from the catalytic site, at sites involved in the first enzyme/substrate interaction. In addition, it incorporates a new mechanism of enzyme activity regulation that could be fundamental to search for new activity-modulating sites or for the design of mutants with modified enzyme parameters.