Carotenoids are a group of isoprenoid pigments naturally synthesized by plants and microorganisms, which are applied industrially in food, cosmetic, and pharmaceutical product formulations. In ...addition to their use as coloring agents, carotenoids have been proposed as health additives, being able to prevent cancer, macular degradation, and cataracts. Moreover, carotenoids may also protect cells against oxidative damage, acting as an antioxidant agent. Considering the interest in greener and sustainable industrial processing, the search for natural carotenoids has increased over the last few decades. In particular, it has been suggested that the use of bioprocessing technologies can improve carotenoid production yields or, as a minimum, increase the efficiency of currently used production processes. Thus, this review provides a short but comprehensive overview of the recent biotechnological developments in carotenoid production using microorganisms. The hot topics in the field are properly addressed, from carotenoid biosynthesis to the current technologies involved in their extraction, and even highlighting the recent advances in the marketing and application of “microbial” carotenoids. It is expected that this review will improve the knowledge and understanding of the most appropriate and economic strategies for a biotechnological production of carotenoids.
Fluorescent proteins have many applications as biomarkers and biosensors in the medical and biological fields. Their success was largely supported by modifications of the first isolated fluorescent ...protein, the wild-type Green Fluorescent Protein (wtGFP), which allowed the development of improved variants such as the Enhanced GFP (EGFP). The first reports on EGFP indicated that the protein presented a single form and fluorescence peak, in contrast to the two conformations observed in wtGFP. However, after experimental determination of the crystalline structure of EGFP, two conformations were found, generating questions regarding the relationship between EGFP structure and its spectral characteristics. To resolve the controversy, this study evaluated EGFP 3D fluorescence spectra at lower wavelengths and under distinct conditions (different concentrations, pH and temperatures), revealing the existence of a second fluorescence peak for this protein. It was possible to confirm that the new peak was not a reflection of the intrinsic fluorescence of proteins or an artefact from the 3D fluorescence spectroscopy. It was also shown that the second peak is pH dependent, sensitive to high temperatures and linearly related to EGFP concentration, confirming a direct relationship between the new fluorescence peak and EGFP protein structure. In addition to the revelation of the new EGFP fluorescence peak, this study demonstrated that 3D fluorescence can be used as powerful technique in the discovery of other elusive fluorophores.
Three-dimensional fluorescence spectroscopy as a powerful tool to identify a new fluorescence peak of Enhanced Green Fluorescent Protein (EGFP).
We evaluated the biogeographic factors that influence composition and diversity of the Neotropical otter diet. We also tested whether
Lontra longicaudis
selected fish in its diet per their ...availability. We collected data from 41 studies, covering most of the otter’s distribution, 32 of which had prey identified to family level. Fish was the main prey for
L. longicaudis
throughout the species range. However, its frequency was higher in sites farther from the coast and at higher slopes. Alternative prey items compensate the decrease in fish. Temperature seasonality, slope, and distance to coast were the most important factors determining the diversity of the otter’s diet. Populations in areas with more variation in temperature and/or closer to the coast showed a more diverse diet with an increase of alternative prey. Trophic diversity was positively correlated with temperature seasonality but negatively correlated with the slope and distance to coast. As an opportunist, semiaquatic predator with a wide distribution range,
L. longicaudis
has a flexible diet, and this plasticity might be explained by the wide range of habitats occupied by otters. Furthermore, Neotropical otter diet was also found to be more similar in geographically closer areas, as well as in similar environment types.
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•The production of carotenoids by Rhodotorula glutinis were optimized applying two statistical designs.•R. glutinis produced simultaneously carotenoids (β-carotene, torularhodin and ...torulene) and lipids.•Combined supplementation with organic and inorganic nitrogen sources have a key role in the production of carotenoids.•The production yield of biomass and carotenoids were increased in the stirred-tank bioreactor.
Rhodotorula strains can produce industrial valuable bioproducts. In this work, the production of carotenoids, and lipids by Rhodotorula glutinis using different nitrogen sources was evaluated. Two statistical experimental designs were applied to improve carotenoid production: a first 25−1 fractional factorial design evaluating the influence of independent variables pH, nitrogen source, glucose, KH2PO4 and MgSO4 concentration; a second 22 central factorial design to optimize the effect of pH and nitrogen sources. After the optimization using two statistical designs, a culture mediium composed of (in g/L) glucose (10), asparagine (10), NH4NO3 (4), KH2PO4 (0.52), MgSO4 (0.52) was found as the best for the production of carotenoids at a pH 5 and 30 °C. The best bioprocess was scaled-up to a 5 L stirred-tank bioreactor. The change to a bioreactor allowed to improve aeration and agitation conditions, and consequently, increasing the production yields (m/v) in, approximately, 25.83 %, 11.88 %, 24.50 % and 10.32 % for β-carotene, torularhodin, torulene and lipids, respectively. The combined supplementation of the culture media with both organic (asparagine) and inorganic nitrogen (ammonium nitrate) sources was primordial for enhancing the carotenogenesis. R. glutinis are very efficient in the production of valuable carotenoids and lipids, presenting high potential of yeast for the industrial production of more sustainable cosmetic, pharmaceutical, and food products.
This work presents a novel approach for tailoring molecularly imprinted polymers (MIPs) with a preliminary stage of atom transfer radical polymerization (ATRP), for a more precise definition of the ...imprinted cavity. A well-defined copolymer of acrylamide and
,
'-methylenebisacrylamide (PAAm-co-PMBAm) was synthesized by ATRP and applied to gold electrodes with the template, followed by a crosslinking reaction. The template was removed from the polymer matrix by enzymatic/chemical action. The surface modifications were monitored via electrochemical impedance spectroscopy (EIS), having the MIP polymer as a non-conducting film designed with affinity sites for CA15-3. The resulting biosensor exhibited a linear response to CA15-3 log concentrations from 0.001 to 100 U/mL in PBS or in diluted fetal bovine serum (1000×) in PBS. Compared to the polyacrylamide (PAAm) MIP from conventional free-radical polymerization, the ATRP-based MIP extended the biosensor's dynamic linear range 10-fold, improving low concentration detection, and enhanced the signal reproducibility across units. The biosensor demonstrated good sensitivity and selectivity. Overall, the work described confirmed that the process of radical polymerization to build an MIP material influences the detection capacity for the target substance and the reproducibility among different biosensor units. Extending this approach to other cancer biomarkers, the methodology presented could open doors to a new generation of MIP-based biosensors for point-of-care disease diagnosis.
Biopharmaceuticals have allowed the control of previously untreatable diseases. However, their low solubility and stability still hinder their application, transport, and storage. Hence, researchers ...have applied different compounds to preserve and enhance the delivery of biopharmaceuticals, such as ionic liquids (ILs) and deep eutectic solvents (DESs). Although the biopharmaceutical industry can employ various substances for enhancing formulations, their effect will change depending on the properties of the target biomolecule and environmental conditions. Hence, this review organized the current state-of-the-art on the application of ILs and DESs to stabilize biopharmaceuticals, considering the properties of the biomolecules, ILs, and DESs classes, concentration range, types of stability, and effect. We also provided a critical discussion regarding the potential utilization of ILs and DESs in pharmaceutical formulations, considering the restrictions in this field, as well as the advantages and drawbacks of these substances for medical applications. Overall, the most applied IL and DES classes for stabilizing biopharmaceuticals were cholinium-, imidazolium-, and ammonium-based, with cholinium ILs also employed to improve their delivery. Interestingly, dilute and concentrated ILs and DESs solutions presented similar results regarding the stabilization of biopharmaceuticals. With additional investigation, ILs and DESs have the potential to overcome current challenges in biopharmaceutical formulation.
•Most Ionic liquids (ILs) and deep eutectic solvents (DESs) improved the stability or delivery of biopharmaceuticals.•The predominant classes of ILs and DESs reported were cholinium-, imidazolium- and ammonium-based.•Dilute and concentrated solutions of ILs and DESs presented similar results.•Cholinium ILs and DESs improved the delivery of biopharmaceuticals.•Despite market expansion, biopharmaceutical complexities endure, demanding continuous R&D to address formulation issues.
•Control of the biomolecules partitioning using ILs as co-surfactants in AMTPs.•Imidazolium-based ILs and fermented broth increase the AMTPs’ cloud point temperature.•Increasing the colorants ...partitioning to the surfactant rich-phase with the addition of high ILs’ concentration.•Temperature strongly influences the partitioning of colorants in ILs-based AMPTs.•Higher temperatures enlarge the partitioning of red colorants into surfactant-rich phase.
Over the last few years, synthetic colorants have been increasingly replaced by natural alternatives that cause fewer adverse health effects and can show biological activity. These biocompounds can be produced by fermentative processes using microorganisms such as bacteria and filamentous fungi. However, the biocompounds must be extracted from the fermented broth before industrial application. Thus, the development of effective techniques for the extraction and purification of biocolorants with high recovery is of great interest. The main goal of this study was to evaluate the recovery of natural colorants produced by Talaromyces amestolkiae using aqueous micellar two-phase system (AMTPS) composed of Triton X-114 and Ionic Liquids (ILs) from the imidazolium family as co-surfactants. Preliminary experiments were performed to determine the binodal curves in both presence and absence of fermented broth containing the red colorants. Subsequently, a set of partitioning experiments was performed using the developed systems. Under all conditions, the red colorants were partitioned mainly into the micelle-rich phase. The systems with low concentrations of Triton X-114 and ILs showed high partition coefficients. The system (0.1wt% C10mimCl+3wt% Triton X-114) with the highest partition coefficient (KRC=6.12) was used to study the effect of temperature on partitioning. The partitioning of the red colorants was strongly influenced by the temperature, and KRC increased to 24.71 at 45°C. The results of this study demonstrate that the partitioning of biomolecules can be controlled by the presence of ILs, mainly by electrostatic interactions, or by the proper adjustment of the partitioning temperature.