New approaches for modulation of alginate-chitosan delivery properties.

Alginate is a biopolymer widely used on delivery systems when bioactive protection at acidic pH is required, while chitosan can enhance mucoadhesion and controlled release at alkaline pHs. In this work, alginate ionotropic gelation and electrostatic complexation to chitosan were evaluated concomitantly or in a two-step approach to improve the delivery properties of systems in different pHs. The effect of pH on alginate gelation and chitosan interactions were also evaluated. Alginate microspheres were prepared by ionotropic gelation in CaCl2 at different pH values (2.5 and 6.0) by extrusion. Complexation with chitosan was carried out during alginate ionotropic gelation (one-step approach) or after alginate gel formation (two-step approach). Alginate microparticles without chitosan showed larger pores and lower mechanical strength. Extruded microspheres at pH 6.0 were more stable to pH and showed smaller pores than the formed at pH 2.5. One-step production retained a large amount of bioactive at pH 7.0 and resulted in lower release at the pH of intestinal digestion. The two-step approach retained less amount of bioactive but confer more protection to the pH of the stomach phase and higher release in pH of the intestinal phase than one-step samples. These results indicate that the formation of alginate gels by ionotropic gelation followed by the complexation with chitosan (in two-step) is promising for the transport and delivery of bioactives into intestinal conditions, whereas the ionotropic gelation concomitantly to electrostatic complexation (one-step approach) is indicated to the delivery of bioactives into lower pH environments.

Antimicrobial activity of lactoferrin-chitosan-gellan nanoparticles and their influence on strawberry preservation.

Lactoferrin (L), chitosan (C) and gellan (G) nanoparticles were produced by electrostatic complexation, aiming to enhance the antimicrobial characteristics of these compounds. Binary complexes (lactoferrin-gellan and chitosan-gellan) and ternary complexes (lactoferrin-chitosan-gellan) were studied in eight different proportions of biopolymers. The antimicrobial activity of nanoparticles against S. aureus was compared with that of pure biopolymers and related to nanoparticle size, charge density and morphology. Those with a 4.5L:4.5C:1G ratio presented the highest positive charge density (+57.90 ± 1.50) mV and smaller hydrodynamic diameter (53.53 ± 2.06) nm, which resulted in the highest antimicrobial action (minimum inhibitory concentration of 0.0117 mg/ml). When applied to a coating of fresh strawberries, the nanoparticles were effective in preserving their physicochemical properties, especially in the presence of carboxymethylcellulose that enhanced the adhesion of particles to the fruits. In this study, the antimicrobial action of nanoparticles was better than that of lactoferrin and pure chitosan alone, proving that the antimicrobial properties of such biopolymers were enhanced due to aggregation at the nanoscale. The nanoparticles produced have outstanding application potential as natural food preservers.

Synthesis, characterization and application of antibacterial lactoferrin nanoparticles.

Lactoferrin (L) and gellan gum (G) nanoparticles were produced in different biopolymer proportions through electrostatic complexation to enhance the antimicrobial properties of lactoferrin. The nanoparticles were characterized according to size, charge density, morphology and antimicrobial activity against S. aureus and E. coli, in two different broths to show the effect of the broth composition on the nanoparticle activity. The 9L:1G particles showed the highest positive zeta potential (+21.20 mV) and reduced diameter (92.03 nm) which resulted in a minimum inhibitory concentration six times smaller (0.3 mg/ml) than pure lactoferrin (2 mg/ml). However, the bacteriostatic action of nanoparticles was inhibited in the presence of divalent cations. When applied to strawberries as a coating, lactoferrin nanoparticles extended fruit shelf-life up to 6 days in the presence of carboxymethylcellulose (CMC). Therefore, lactoferrin-gellan gum complexation was proved to be a promising tool to enhance lactoferrin antimicrobial action and broaden its application as a food preserver.

Structural and mechanical properties of organogels: Role of oil and gelator molecular structure.

This work aims at evaluating the influence of oil and gelator structure on organogels' properties through rheological measurements, polarized microscopy and small-angle X-ray scattering (SAXS). Four different food-grade gelators (glyceryl tristearate - GT; sorbitan tristearate - ST; sorbitan monostearate - SM and glyceryl monostearate - GM) were tested in medium-chain triglyceride and high oleic sunflower (MCT and LCT, respectively) oil phases. Organogels were prepared by mixing the oil phase and gelator at different concentrations (5, 10, 15, 20 and 25%) at 80°C during 30min. All organogels presented birefringence confirming the formation of a crystalline structure that changed with the increase of the gelator concentration. Through the evaluation of SAXS peaks it has been confirmed that all structures were organized as lamellas but with different d-spacing values. These particularities at micro- and nanoscale level lead to differences in rheological properties of organogels. Results showed that the oil type (i.e. medium- and long-chain triglyceride) and hydrophilic head of gelators (i.e. sorbitan versus glyceryl) exert influence on the organogels physical properties, but the presence of monostearate leads to the formation of stronger organogels. Moreover, gels produced with LCT were stronger and gelled at lower organogelator concentration than MCT.

Breaking oil-in-water emulsions stabilized by yeast.

Several biotechnological processes can show an undesirable formation of emulsions making difficult phase separation and product recovery. The breakup of oil-in-water emulsions stabilized by yeast was studied using different physical and chemical methods. These emulsions were composed by deionized water, hexadecane and commercial yeast (Saccharomyces cerevisiae). The stability of the emulsions was evaluated varying the yeast concentration from 7.47 to 22.11% (w/w) and the phases obtained after gravity separation were evaluated on chemical composition, droplet size distribution, rheological behavior and optical microscopy. The cream phase showed kinetic stability attributed to mechanisms as electrostatic repulsion between the droplets, a possible Pickering-type stabilization and the viscoelastic properties of the concentrated emulsion. Oil recovery from cream phase was performed using gravity separation, centrifugation, heating and addition of demulsifier agents (alcohols and magnetic nanoparticles). Long centrifugation time and high centrifugal forces (2 h/150,000×g) were necessary to obtain a complete oil recovery. The heat treatment (60°C) was not enough to promote a satisfactory oil separation. Addition of alcohols followed by centrifugation enhanced oil recovery: butanol addition allowed almost complete phase separation of the emulsion while ethanol addition resulted in 84% of oil recovery. Implementation of this method, however, would require additional steps for solvent separation. Addition of charged magnetic nanoparticles was effective by interacting electrostatically with the interface, resulting in emulsion destabilization under a magnetic field. This method reached almost 96% of oil recovery and it was potentially advantageous since no additional steps might be necessary for further purifying the recovered oil.

Microbial advanced biofuels production: overcoming emulsification challenges for large-scale operation.

Isoprenoids and alkanes produced and secreted by microorganisms are emerging as an alternative biofuel for diesel and jet fuel replacements. In a similar way as for other bioprocesses comprising an organic liquid phase, the presence of microorganisms, medium composition, and process conditions may result in emulsion formation during fermentation, hindering product recovery. At the same time, a low-cost production process overcoming this challenge is required to make these advanced biofuels a feasible alternative. We review the main mechanisms and causes of emulsion formation during fermentation, because a better understanding on the microscale can give insights into how to improve large-scale processes and the process technology options that can address these challenges.