Artigos - Bioprocessos e Biotecnologia - Araraquara

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https://hdl.handle.net/11449/132948

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Ionic liquids and deep eutectic solvents for the stabilization of biopharmaceuticals: a review
(Elsevier, 2024-01-08) Veríssimo, Nathalia Vieira Porphirio [UNESP]; Mussagy, Cassamo Usemane; Bento, Heitor Buzetti Simões [UNESP]; Pereira, Jorge Fernando Brandão; Santos-Ebinuma, Valéria de Carvalho [UNESP]; Universidade de São Paulo; Pontificia Universidad Católica de Valparaíso; Universidade de Coimbra
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.
Effect of amphiphilic ionic liquids on the colorimetric properties of polyketides colorants
(Elsevier, 2022-07-16) Veríssimo, Nathalia Vieira [UNESP]; Nakamura, Cecília Naomi [UNESP]; Oliveira, Fernanda de [UNESP]; Kuhn, Bruna L.; Frizzo, Clarissa P.; Pereira, Jorge F. B. [UNESP]; Santos-Ebinuma, Valéria C. [UNESP]; Universidade Estadual Paulista (Unesp)
Consumers are currently pressuring the food industries to replace synthetic colorants with their natural counterparts. However, natural colorants still represent less than a third of food coloring additives, as they are more difficult to produce and present application issues due to their lower stability. To find surfactants compatible with natural colorants, this study evaluated the effect of amphiphilic ionic liquids (ILs) aqueous solutions on the colorimetric properties of polyketides colorants (yellow-orange and red color) produced by Talaromyces amestolkiae. Only the IL n-alkyl-3-methylimidazolium bromide ([Cnmim]Br with n = 10 at 2.5 mM preserved the colorimetric and fluorescence characteristics of the fungal colorants, while the other surfactants (e.g., [Cnmim]Br with n = 12, 14, and 16, [P44414]Cl, and CTAB) altered them. Fluorescence was more sensitive to the effect of ILs than their color and absorbance. Furthermore, the red colorant was more affected by the ILs than the yellow-orange chromophores. Hence, this work shows that formulations with [C10mim]Br (up to 2.5 mM) can preserve the properties of T. amestolkiae polyketides colorants, while the other ILs can be used as additives to modulate their color and fluorescence for biosensing applications.
Improving the cost effectiveness of enhanced green fluorescent protein production using recombinant Escherichia coli BL21 (DE3): Decreasing the expression inducer concentration
(Wiley, 2019-04-08) dos Santos, Nathalia Vieira [UNESP]; Pereira, Jorge Fernando Brandão [UNESP]; Pedrolli, Danielle Biscaro [UNESP]; Santos-Ebinuma, Valéria De Carvalho [UNESP]; Valentini, Sandro Roberto [UNESP]; Lopes, Camila [UNESP]; Dupont, Jana [UNESP]; Universidade Estadual Paulista (Unesp)
Green Fluorescent Protein (GFP) is a globular protein used as biosensor and biomarker in medical and industrial fields. However, due to the expensive production costs of expressing proteins using high-cost inducers like isopropyl-β-D-1-thiogalactopyranoside (IPTG), the number of GFP applications are still scarce. This work studied the production of Enhanced GFP (EGFP) using Escherichia coli BL21 (DE3) [pLysS; pET28(a)], aiming to increase its yield and reduce costs. Firstly, the influence of agitation rate, induction time and concentration of IPTG in the production of EGFP were evaluated, but only the first two parameters were significant. Subsequently, aiming to reduce costs related to the use of inducer, the IPTG concentration (0.005, 0.010 and 0.025 mM) was decreased and, interestingly, the production levels were maintained or increased. These results show that a proper choice of production conditions, particularly through the decrease of inducer concentration, is effective to reduce the upstream production costs and guarantee high EGFP expression.
From green to blue economy: Marine biorefineries for a sustainable ocean-based economy
(Royal Chemical Society, 2021-12-07) Veríssimo, Nathalia Vieira; Mussagy, Cassamo Ussemane; Oshiro, Ariane Alves [UNESP]; Mendonça, Carlos Miguel Nóbrega; Santos-Ebinuma, Valéria De Carvalho [UNESP]; Pessoa, Adalberto; Oliveira, Ricardo Pinheiro De Souza; Pereira, Jorge Fernando Brandão [UNESP]; Universidade Estadual Paulista (Unesp); Universidade de São Paulo (USP)
Despite being a vital asset for global sustenance and economy, ocean aquatic ecosystems are in danger due to the effects of incorrect management of their resources, pollution, and climate change. Considering the seafood industry discards half of its fish-product mass in the ocean, a proper valorization of its residues would decrease not only the ocean contamination but also improve the management of marine resources and increase the sector competitiveness. With these goals in mind, ocean-based industries are adopting new sustainable production models, similar to biorefineries, which are effective for waste valorization, namely, converting low-value biomass into commercially relevant by-products. Based on a deeper knowledge of aquatic feedstocks, the development and implementation of a marine biorefinery can be fundamental to consolidate a greenersocioeconomic development, similar to that observed in green chemistry. However, biorefineries are sophisticated multi-step systems with numerous feedstocks and commodities. Therefore, their implementation requires expertise in all stages of manufacturing, in addition to a clear vision of all raw materials, residues, and products. In this sense, with this perspective, we provide an initial overview of the current state-of-the-art on marine biorefineries and the sources and applications of their by-products. Afterward, we suggest how to integrate green chemistry and blue economy principles into ocean-based industries, aiming to support a more sustainable, profitable, and conscious ocean economy. This journal is
Obtaining of xanthan gum impregnated of cellulose microfibrils derived from sugarcane bagasse
(2015) Gomes, Gleice V. P.; Assis, Denilson de J.; Silva, Jania B. A. da; Santos, Valéria de Carvalho [UNESP]; Costa, Larissa A. S.; Druzian, Janice I.; Universidade Estadual Paulista (Unesp); Universidade Federal da Bahia (UFBA); Universidade Federal do Recôncavo Baiano; Instituto Federal Baiano
Xanthan gum (XG) is a microbial exopolysaccharide that occupies a prominent place in the market because of its unique properties compared with other microbial gums. When lignocellulosic materials such as sugarcane bagasse (SCB) are used as a carbon source, the degradation of the SCB material can be partial, retaining microfibrils of cellulose in the fermentation broth. These microfibrils can be incorporated into the XG structure. The aim of this work was to select the best concentration of SCB to produce XG and to analyze the characteristics of the biopolymer obtained. The XG produced in the best condition was added to biodegradable films as a reinforcing agent. The SCB was evaluated at the following concentrations: 2, 4, 6, 8 and 10% w/w. The XG produced with 4% w/w SCB provided the best XG, not only for production but also for polymer characteristics. Visualization with transmission electron microscopy (TEM) established the presence of microfibrils in the polysaccharide structure, and the incorporation of XG with microfibrils provided a flexible film with enhanced mechanical properties.
Estudo de modelo matemático do comportamento oscilatório em processos contínuos de fermentação alcoólica com Zymomonas mobilis
(2014) Gambarato, Bruno Chaboli; Oliveira, Samuel Conceição de [UNESP]; Universidade Estadual Paulista (Unesp)
Neste trabalho, foi estudado o modelo matemático oscilatório proposto por Daugulis et al. (1997) referente a um processo contínuo de fermentação alcoólica, de modo a compreender sua estrutura, variáveis, fenômenos e hipóteses postuladas. O estudo foi realizado de modo a compreender como os mecanismos de inibição por produto e substrato, e como os mecanismos adaptativos do microrganismo são levados em consideração pelo modelo, resultando no comportamento oscilatório desejado. Por fim, foram realizadas simulações com perturbações do tipo degrau na taxa de diluição e na concentração de substrato na alimentação, de modo a extrapolar o comportamento do modelo.
A dual control mechanism synchronizes riboflavin and sulphur metabolism in Bacillus subtilis
(2015-10-22) Pedrolli, Danielle Biscaro [UNESP]; Kühm, Christian; Sévin, Daniel C.; Vockenhuber, Michael P.; Sauer, Uwe; Suess, Beatrix; Mack, Matthias; Universidade Estadual Paulista (Unesp); Department of Biotechnology, Institute for Technical Microbiology, Hochschule Mannheim, 68163 Mannheim, Germany;; Institute of Molecular Systems Biology, Eidgenössische Technische Hochschule (ETH) Zürich, 8093 Zürich, Switzerland;; Department of Biology, Technical University Darmstadt, 64287 Darmstadt, Germany.; Department of Biotechnology, Institute for Technical Microbiology, Hochschule Mannheim, 68163 Mannheim, Germany; m.mack@hs-mannheim.de.
Flavin mononucleotide (FMN) riboswitches are genetic elements, which in many bacteria control genes responsible for biosynthesis and/or transport of riboflavin (rib genes). Cytoplasmic riboflavin is rapidly and almost completely converted to FMN by flavokinases. When cytoplasmic levels of FMN are sufficient (high levels), FMN binding to FMN riboswitches leads to a reduction of rib gene expression. We report here that the protein RibR counteracts the FMN-induced turn-off activities of both FMN riboswitches in Bacillus subtilis, allowing rib gene expression even in the presence of high levels of FMN. The reason for this secondary metabolic control by RibR is to couple sulfur metabolism with riboflavin metabolism.
Benzene solubility in ionic liquids: working toward an understanding of liquid clathrate formation
(Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim, 2014) Pereira, Jorge Fernando Brandão [UNESP]; Flores, Luis A.; Wang, Hui; Rogers, Robin D.; Universidade Estadual Paulista (Unesp); University of Alabama
The solubility of benzene in 15 imidazolium, pyrrolidinium, pyridinium, and piperidinium ionic liquids has been determined; the resulting, benzene-saturated ionic liquid solutions, also known as liquid clathrates, were examined with (1) H and (19) F nuclear magnetic resonance spectroscopy to try and understand the molecular interactions that control liquid clathrate formation. The results suggest that benzene interacts primarily with the cation of the ionic liquid, and that liquid clathrate formation (and benzene solubility) is controlled by the strength of the cation-anion interactions, that is, the stronger the cation-anion interaction, the lower the benzene solubility. Other factors that were determined to be important in the final amount of benzene in any given liquid clathrate phase included attractive interactions between the anion and benzene (when significant), and larger steric or free volume demands of the ions, both of which lead to greater benzene solubility.
Adsorption of immunoglobulin Y in supermacroporous continuous cryogel with immobilized Cu2+ ions
(Elsevier B.V., 2015-05-22) Silva Junior, Willer Ferreira da; Cano, Rodrigo; Totola, Antonio Helvecio; Carvalho, Lorendane Milena de; Cerri, Marcel Otavio [UNESP]; Reis Coimbra, Jane Sella dos; Pinto de Carvalho, Gleidson Giordano; Aparecida de Carvalho, Bruna Mara; Univ Fed Sao Joao del Rei; Universidade Federal de Viçosa (UFV); Universidade Estadual Paulista (Unesp); Universidade Federal da Bahia (UFBA); Universidade Federal de Minas Gerais (UFMG)
The influence of temperature, pH and ionic strength on the adsorption of Immunoglobulin Y (IgY) in IDA-Cu2+ cryogel system was studied by batch equilibrium Measurements. The adsorptive equilibrium data were obtained at 17 and 27 degrees C, pH 5.0 and 6.5, and at ionic strength of 50 and 200 mmol L-1 NaCl. Langmuir, Freundlich and Langmuir-Freundlich models were fitted to equilibrium data, while the enthalpy of adsorption of IgY in IDA-Cu2+ cryogel system was calculated through Van't Hoff analysis. The binding of IgY on cryogel was stronger at 27 degrees C and lowest pH and ionic strength values, with apparent maximum adsorption capacity of 27 mg g(-1). The adsorption of protein in the resin was spontaneous in all analyzed cases. The results provide valuable information to enable the improvement of IgY purification processes. (C) 2015 Elsevier BY. All rights reserved.
Continuous enzymatic interesterification of milkfat with soybean oil produces a highly spreadable product rich in polyunsaturated fatty acids
(Wiley-Blackwell, 2015-05-01) Paula, Ariela Veloso de [UNESP]; Nunes, Gisele Fátima Morais; Osório, Natália M.; Santos, Júlio César dos; Castro, Heizir Ferreira de; Ferreira-Dias, Suzana; Universidade Estadual Paulista (Unesp); Universidade de São Paulo (USP); University of Lisbon
The aim of this study was to tune the physical properties of milkfat by enzymatic interesterification with soybean oil in a continuous fluidized bed reactor (FBR) to obtain healthy interesterified fat blends having suitable texture properties for the food industry. The immobilized commercial non-regioselective Candida antarctica lipase (Novozym (R) 435) and sn1,3-regioselective Rhizopus oryzae lipase, immobilized in an organic-inorganic hybridmatrix of polysiloxane-polyvinyl alcohol, were used as biocatalysts in a FBR. The minimum value of the ascendant flow of the medium for allowing fluidization in the system was 3.13 mL.min(-1). The reaction was evaluated in terms of the interesterification yield (IY), consistency values and solid fat content (SFC). The IY values of 10.50 +/- 1.64% and of 5.70 +/- 1.46% were attained for Novozym (R) 435 and for immobilized R. oryzae lipase, respectively. The consistency of the initial 65: 35 milkfat/soybean oil mixture (1000 gf/cm(2)) decreased to 732.35 +/- 75.30 gf/cm(2) and to 478.02 +/- 71.80 gf/cm(2) in interesterified blends catalyzed by Novozym (R) 435 or by R. oryzae lipase, respectively. SFC was considered an inadequate parameter for following the interesterification of this blend formulation because the values were similar for initial and for interesterified blends. Free fatty acid levels of 1.5%, the non-notable deactivation of Novozym (R) 435 and a half-life of 190 h for R. oryzae lipase were observed during the operation time.
Effect of temperature on salt-salt aqueous biphasic systems: manifestations of upper critical solution temperature
(Springer, 2015-04-01) Dilip, Meghna; Bridges, Nicholas J.; Rodriguez, Hector; Pereira, Jorge Fernando Brandão [UNESP]; Rogers, Robin D.; University Alabama; Worcester State University; University Santiago de Compostela; Universidade Estadual Paulista (Unesp)
Water-miscible ionic liquids (ILs) may be salted out using kosmotropic salts such as potassium phosphate (K3PO4) to form salt-salt aqueous biphasic systems (ABS). The effect of temperature on these systems has been studied using phase diagrams and it is observed that the degree of binodal shift decreases (requiring lower IL and kosmotropic salt concentrations) with the increase of temperature following the trend [C(4)mim]Cl > [C(4)py]Cl > [C(4)mmim] Cl > [N-4444]Cl. This trend can be correlated with the decreasing hydrogen bonding abilities of each salt. The phase behavior was also interpreted on the basis of critical solution temperature behavior of pure aqueous ionic liquid solutions. Additionally, the distribution of alcohols in these systems was studied as a function of temperature and it was found that the distribution ratios did not change with changes in temperature. The Gibbs energy of transfer of a methylene group in these systems and correlation to tie-line length was also determined. It was concluded that while the miscibility of alcohols increases in the ILs with increasing temperature, phase divergence in the aqueous biphasic system decreases, and thus these competing forces tend to cancel each other out for small polar molecules. A comparison is provided for the response to temperature in the currently studied salt-salt systems and analogous ABS formed by the addition of hydrophilic polymers to kosmotropic salts (polymer-salt) or other polymers (polymer-polymer).
Energy recovery from agro-industrial wastewaters through biohydrogen production: kinetic evaluation and technological feasibility
(Elsevier B.V., 2015-03-01) Lucas, S. D. M.; Peixoto, G. [UNESP]; Mockaitis, G.; Zaiat, M.; Gomes, S. D.; Western Parana State Univ; Universidade Estadual Paulista (Unesp); Universidade de São Paulo (USP); Natl Res Council Canada NRC
Biohydrogen production from cassava, dairy and citrus processing wastewaters (WWs) without nutritional supplementation was evaluated in anaerobic single-batch reactors at 37 degrees C for 70 h. Hydrogen production from cassava, dairy and citrus WW was 31.41, 37.25 and 28.95 mL g(-1) of chemical oxygen demand (COD). The kinetic parameters indicated that Hy production rates for cassava processing WW (032 mL h(-1)) and dairy WW (0.31 mL h(-1)) were similar, whereas citrus processing WW exhibited the highest value (0.59 mL h(-1)). The carbohydrate degradation rate (k(1)(App)) was highest for dairy WW (0.045 h(-1)), but the most efficient overall conversion of organic matter to Hy (k(2)(App)) was observed with cassava WW (0.014 h(-1)). The rate of conversion of the organic matter of the cassava WW together with its ready availability resulted in a recovery of 0.59 10(9) MJ year(-1), the highest in this study. Cassava WW showed the highest hydrogen production potential (97.9 mL), when compared with dairy (76.1 mL) and citrus WIN (66.6 mL). The economic estimation based on the gasoline energetic equivalent indicated that a single process of H-2 production allowed the maximum economic yield of US$ 0.009 cents L-1 WW. Alternatively, a sequential hydrogen and methane process could increase energy recoveries and economic yields to values near 10.48 kJ g(-1) COD and US$ 0.61 cents L-1 WW (US$ 6.10 m(-3) WW). (C) 2014 Elsevier Ltd. All rights reserved.
Synthesis of Structured Lipids by Enzymatic Interesterification of Milkfat and Soybean Oil in a Basket-Type Stirred Tank Reactor
(Amer Chemical Soc, 2015-02-18) Paula, Ariela Veloso de [UNESP]; Nunes, Gisele F. M.; Castro, Heizir F. de; Santos, Julio C.; Universidade Estadual Paulista (Unesp); Ctr Fed Educ Tecnol Minas Gerais CEFET MG; Universidade de São Paulo (USP)
Lipase from Rhizopus oryzae immobilized on polysiloxane-poly(vinyl alcohol) (SiO2-PVA) was used to study the interesterification reaction of the milkfat with soybean oil in a stirred tank reactor (STR) containing baskets for the immobilized enzyme retention in two different configurations: central or lateral. The progress of the reaction was followed by determining free fatty acids, composition in triacylglycerols (TAGs), and consistency. The central basket was chosen for assessing the biocatalyst operational stability by running 10 consecutive batch assays lasting 6 h each. Non-notable deactivation of the biocatalyst was observed during the total operation time (60 h). The potential of the evaluated system to make the milkfat-based fat more spreadable under cooling temperature and with lower saturated fatty acids content was demonstrated in this study. Both evaluated basket reactor designs have shown potential to be used in interesterification reactions of industrial interest.

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