Desenvolvimento e aplicações de um biorreator airlift com circulação externa

Dissertação (Mestrado) - Universidade Federal de Santa Catarina, Centro TecnologicoUm reator airlíft com circlação externa construído em vidro foi desenvolvido utilizando relações geométricas sugeridas na literatura e dados obtidos através de estudos preliminares utilizando um biorreator de PVC. O comportamento hidrodinâmico do sistema foi estudado através da determinação do gas holdup e da velocidade superficial do líquido. Foi proposto um modelo para a determinação do coeficiente volumétrico de transferência de massa neste reator. Os resultados tiveram uma boa concordância quando comparados com a literatura. Verificou-se a presença de dois regimes de bolhas, homogêneo e heterogêneo e uma zona de transição entre os dois regimes. A velocidade de circulação do líquido mostrou ser influenciada pela diferença de gas holdup do riser e do downcomer, pelo gas holdup no riser e pela altura do biorreator. O desempenho do biorreator airlift foi avaliado através da realização de processos fermentativos em batelada com dois microrganismos: Saccharomyces cerevisiae e Alcaligenes eutrophus. Os resultados obtidos correspondem aos valores encontrados em fermentadores agitados clássicos (STR) utilizando os mesmos microrganismos, meios de cultura e fatores fisico-químicos. Portanto, conclui-se que os fermentadores airlift, mesmo que em escala laboratorial, atende as necessidades de transferência de massa e de agitação para os processos estudados

Fast accumulation of few polyhedra mutants during passage of a Spodoptera frugiperda multicapsid nucleopolyhedrovirus (Baculoviridae) in Sf9 cell cultures

Baculoviruses are a group of viruses that infect invertebrates and that have been used worldwide as a biopesticide against several insect pests of the Order Lepidoptera. In Brazil, the baculovirus Spodoptera frugiperda multicapsid nucleopolyhedrovirus (SfMNPV, Baculoviridae) has been used experimentally to control S. frugiperda (Lepidoptera: Noctuidae), an important insect pest of corn (maize) fields and other crops. Baculoviruses can be produced either in insect larvae or in cell culture bioreactors. A major limitation to the in vitro production of baculoviruses is the rapid generation of mutants when the virus undergoes passages in cell culture. In order to evaluate the potential of in vitro methods of producing SfMNPV on a large-scale, we have multiplied a Brazilian isolate of this virus in cell culture. Extensive formation of few polyhedra mutants was observed after only two passages in Sf9 cells

Techno-functionality of fisetin-enriched yoghurt fermented with Lactobacillus acidophilus bio-capsules produced via osmoporation

International audienceOsmoporation is a novel encapsulation approach for bioactive compounds based on the osmoresistance mechanisms of microbial cells. To the best of our knowledge, this is the first study investigating the production of fisetin-enriched yoghurt using Lactobacillus acidophilus-based bio-capsules via osmoporation as the starter culture. Results showed that the milk acidification with fisetin-loaded L. acidophilus progressed at a slower pace due to complex mechanisms induced by osmoporation and internalized fisetin. Milk fermentation using fisetin bio-capsules reached a maximum acidification rate of 0.18 pH units/h after 23 h and pH 4.6 was achieved after 32 h. Besides, the antioxidant activity of yoghurts produced with fisetin bio-capsules did not change during cold storage, while the antioxidant activity of yoghurt produced with non-encapsulated fisetin was reduced by 2.5-fold after 28 days. Overall, this study shows that fisetin osmoporation using L. acidophilus is a versatile encapsulation bioprocess that enables the delivery of preserved phytoactives into fermented foods like yoghurt. This strategy has the potential to be extended to other applications in the dairy industry using lactic acid bacteria as both the encapsulation matrix and fermentation agent

Osmoporation is a versatile technique to encapsulate fisetin using the probiotic bacteria Lactobacillus acidophilus.

International audienceThe objective of this study was to evaluate the performance of Lactobacillus acidophilus cells as a novel encapsulating carrier for fisetin via osmoporation. Initially, the effects of osmotic pressure and initial fisetin concentration on the performance of the osmoporation process were evaluated. The best results were achieved when 15 MPa was applied, while the maximum loading capacity was reached when fisetin concentration of 2.0 mg·mL-1 was used. For these conditions, the cell viability, encapsulation efficiency (EE), and encapsulated fisetin content (EF) were 72%, 28%, and 0.990 mg, respectively. Further, the encapsulation was confirmed by Fourier transform-infrared (FT-IR), differential scanning calorimetry (DSC), and X-ray diffraction (XRD) analysis. DSC thermograms revealed an increase of 40 °C in the melting point of fisetin after encapsulation. In addition, the enhancement of fisetin bioaccessibility by osmoporated biocapsules is shown for the first time in the literature. When the fisetin biocapsules were subjected to in vitro gastrointestinal digestion, 99.6% of the encapsulated content were retained through the gastric stage and 45.5% were released during the intestinal stage, despite no active cells were detected during simulated digestion. These results suggest that alive cells are required for an effective osmoporation-assisted encapsulation process; however, osmoporated biocapsules can efficiently protect and preserve labile compounds, independently of their activity. Overall, this study demonstrated that osmoporation using probiotic L. acidophilus is a simple, versatile, and efficient technique to encapsulate and deliver lipophilic fisetin for food applications. KEY POINTS : •Fisetin is efficiently encapsulated into L. acidophilus via osmoporation. •Fisetin bioaccessibility is improved by osmoporation into L. acidophilus. •Release mechanisms of osmoporation carriers are independent of the cell activity

Sonoprocessing enhances the stabilization of fisetin by encapsulation in Saccharomyces cerevisiae cells

International audienceThe objective of this study was to investigate for the first time the role of S. cerevisiae natural barriers and endogenous cytoplasmatic bodies on the stabilization of fisetin encapsulated via sonoprocessing coupled to freeze-drying (FD) or spray drying (SD). Both protocols of encapsulation improved the resistance of fisetin against thermal treatments (between 60 and 150 °C) and photochemical-induced deterioration (light exposition for 60 days) compared to non-encapsulated fisetin (antioxidant activity retention of approximately 55% and 90%, respectively). When stored under constant relative humidity (from 32.8 to 90%) for 60 days, yeast carriers improved the half-life time of fisetin by up to 4-fold. Spray dried particles were smaller (4.9 μm) and showed higher fisetin release after simulated gastrointestinal digestion (55.7%) when compared to FD. Freeze-dried particles, in turn, tended to agglomerate more than SD (zeta potential -19.7 mV), resulting in reduced loading features (6.3 mg/g) and less efficient protection of fisetin to heat, photo, and moisture-induced deterioration. Overall, spray-dried sonoprocessed fisetin capsules are an efficient way to preserve fisetin against harsh conditions. Altogether, this report shows that sonoprocessing coupled to drying is an efficient, creative, and straightforward route to protect and deliver lipophilic fisetin using yeast capsules for food applications

Fisetin yeast-based bio-capsules via osmoporation: effects of process variables on the encapsulation efficiency and internalized fisetin content

International audienceOsmoporation is an innovative method that can be used with food-grade yeast cells of Saccharomyces cerevisiae as natural encapsulating matrices. This technique overcomes barriers that difficult encapsulation and enables the internalization of fragile bioactive molecules such as fisetin into yeasts. In the present study, we assessed the effects of concentration, osmotic pressure, and temperature on the encapsulation efficiency (EE) and internalized fisetin content (IF). Two different quantification strategies were investigated: direct extraction (DE) without cell washing or freeze-drying steps and indirect extraction (IE) performed after washings with ethanol and freeze-drying. Our results showed that osmoporation improved EE (33 %) and IF (1.199 mg). The best experimental conditions were found by using DE. High-resolution images showed that the yeast cell envelope was preserved during osmoporation at 30 MPa and 84 % of yeast cells remained viable after treatment. Washing cells with organic solvent led to decreased EE (0.65 %) and IF (0.023 mg). This was probably due to either damages caused to yeast cell envelope or fisetin dragged out of cell. Overall, the results demonstrated the adequacy and relevant biotechnological potential of yeasts as encapsulating matrices for hydrophobic compounds. This fresh biotechnological approach has proven to be a promising tool for the production of bioactive-rich food products

Sonoprocessing is an effective strategy to encapsulate fisetin into Saccharomyces cerevisiae cells

International audienceThe encapsulation of fisetin into S. cerevisiae cells through sonoporation coupled with drying is reported for the first time in the literature. To establish the best conditions to maximize the amount of internalized fisetin, the cell density (5-10% w/v), fisetin concentration (1-3 mg/mL), acoustic energy density (0-333.3 W/L), and drying method (freeze-drying and spray drying) were analyzed through a Box-Behnken experimental design (BBD) coupled with response surface methodology (RSM). Higher encapsulation efficiency (EE) was achieved with a cell density of 10% w/v, while fisetin concentration of 3 mg/mL favored the encapsulation yield (EY) and antioxidant activity (AA). Higher EE (67.7%), EY (25.7 mg/g), and AA (90%) were registered when an acoustic density of 333.3 W/L was used. Furthermore, both drying protocols promoted fisetin encapsulation, but through spray drying, the EE, EY, and AA were 11.5%, 11.1%, and 26.6% higher than via freeze-drying, respectively. This work proved that fully filled biocapsules were produced through sonoprocessing, and their morphology was influenced by the acoustic energy and drying process. Overall, these results open new perspectives for the application of sonoprocessing-assisted encapsulation, paving the way for developing innovative yeast-based delivery systems for lipophilic compounds such as fisetin. KEY POINTS: • Sonoprocessing improves the encapsulation of fisetin into S. cerevisiae cells • Spray drying promotes fisetin loading into yeasts' intracellular space and cavities • Fisetin binding with yeast extracellular agents are favored by freeze-drying

Kinetic analysis of in vitro production of wild-type Spodoptera frugiperda nucleopolyhedrovirus

In this study, the kinetic behavior of Sf9 and Sf21 cells used in the production of a baculovirus biopesticide to control the pest of corn Spodoptera frugiperda was analyzed. Kinetic variables such as maximum specific growth rate, cell productivity, mean rate of infection, as well as the mean rate of occlusion body production were determined during the infection of these cell-lines with the extracellular virus of the S. frugiperda nucleopolyhedrovirus (SfMNPV). The Sf9 cell-line resulted in better viral production results (5.0 x 10 OB/mL) than the Sf21 cell-line (2.5 x 10 OB/mL)