Enzymatic production process of capric acid-rich structured lipids: Development of formulation as a new therapeutic approach.

The present work reports an optimization of the synthesis of MLM-type (medium, long, medium) structured lipids (SL) through an acidolysis reaction of grape seed oil with capric acid catalyzed by Rhizopus oryzae lipase immobilized. At first, tests were carried out by preparing the biocatalysts using enzyme loadings (0.15 to 1 g of enzymatic powder) for each gram of support. Enzyme loading was used 0.3 g of enzymatic powder, and hydrolytic activity of 1860 ± 23.4 IU/g was reached. Optimized conditions determined by the Central Composite Rotatable Design (CCRD) revealed that the acidolysis reaction reached approximately 59 % incorporation degree (%ID) after 24 h, in addition to the fact that the biocatalyst could maintain the incorporation degree in five consecutive cycles. From this high incorporation degree, cell viability assays were performed with murine fibroblast cell lines and human cervical adenocarcinoma cell lines. Concerning the cytotoxicity assays, the concentration of MLM-SL to 1.75 and 2 % v/v were able to induce cell death in 56 % and 64 % of adenocarcinoma cells, respectively. Human cervical adenocarcinoma cells showed greater sensitivity to the induction of cell death when using emulsions with MLM-SL > 1.75 % v/v compared to emulsions with lower content indicating a potential for combating carcinogenic cells.

A look into Phaffia rhodozyma biorefinery: From the recovery and fractionation of carotenoids, lipids and proteins to the sustainable manufacturing of biologically active bioplastics.

Carotenoids over-producing yeast has become a focus of interest of the biorefineries, in which the integration of the bioproduction with the following downstream processing units for the recovery and purification of carotenoids and other value-added byproducts is crucial to improve the sustainability and profitability of the overall bioprocess. Aiming the future implementation of Phaffia rhodozyma-based biorefineries, in this work, an integrative process for fractionation of intracellular compounds from P. rhodozyma biomass using non-hazardous bio-based solvents was developed. After one-extraction step, the total amount of astaxanthin, ß-carotene, lipids and proteins recovered was 63.11 µg/gDCW, 42.81 µg/gDCW, 53.75 mg/gDCW and 10.93 mg/g, respectively. The implementation of sequential back-extraction processes and integration with saponification and precipitation operations allowed the efficient fractionation and recovery (% w/w) of astaxanthin (∼72.5 %), ß-carotene âˆ¼90.17 %), proteins (21.04 %) and lipids (23.72 %). After fractionation, the manufacture of carotenoids-based products was demonstrated, through the mixture of carotenoids-rich extracts with bacterial cellulose to obtain biologically active bioplastics.

Advanced methylene blue - nanoemulsions for in vitro photodynamic therapy on oral and cervical human carcinoma.

Photodynamic therapy (PDT) is a therapeutic modality with high contributions in the treatment of cancer. This approach is based on photophysical principles, which presents as a less invasive strategy than conventional therapies. Combined with nanotechnology, the therapy becomes more efficient because nanoparticles (NPs) have advantageous characteristics such as biocompatibility, controlled, and targeted release, promoting solubility and decreasing the toxicity and side effects involved. In this work were developed nanoemulsions containing the methylene blue photosensitizer (MB) (MB/NE) and in the empty form (unloaded/NE). Subsequently, the mentioned nanomaterials were characterized by the measurement of dynamic light scattering (DLS). The MB/NE and unloaded/NE showed appropriate physical and chemical characteristics, with particle size ≤ 200 nm, polydispersity index close to 0.3, and zeta potential exhibiting negative charge, showing stable values during the analysis. The incorporation of the MB did not cause changes in the photophysical profile of the photosensitizer. The quantification performed showed an incorporation rate of 81.9%. Viability studies showed an absence of cytotoxicity for MB/NE in the concentrations of 10-75 µmol·L-1, free MB at the concentration of 75 µmol·L-1, and unloaded NE 47.5% (v/v), presenting viability close to 90%, respectively. PDT in vitro protocols applied to OSCC and HeLa cells showed a decrease in cell viability through only one irradiation, evidencing the photodynamic activity of the formulation when applied to cancer cells. The results obtained were superior to those found in the literature where they use free MB, showing that the association between nanotechnology and PDT optimizes the proposed protocol. From the results obtained, it is possible to indicate that the NE have high stability, with satisfactory physical-chemical parameters, in addition to not presenting cytotoxicity in the tested concentrations, showing their in vitro biocompatibility, in addition to presenting satisfactory effects when combined MB/NE with PDT, showing the potential of MB/NE as a very promising nanostructured photosensitizer for the treatment of some types of cancer.