Fungal Proteins from Sargassum spp. Using Solid-State Fermentation as a Green Bioprocess Strategy.

The development of green technologies and bioprocesses such as solid-state fermentation (SSF) is important for the processing of macroalgae biomass and to reduce the negative effect of Sargassum spp. on marine ecosystems, as well as the production of compounds with high added value such as fungal proteins. In the present study, Sargassum spp. biomass was subjected to hydrothermal pretreatments at different operating temperatures (150, 170, and 190 °C) and pressures (3.75, 6.91, and 11.54 bar) for 50 min, obtaining a glucan-rich substrate (17.99, 23.86, and 25.38 g/100 g d.w., respectively). The results indicate that Sargassum pretreated at a pretreatment temperature of 170 °C was suitable for fungal growth. SSF was performed in packed-bed bioreactors, obtaining the highest protein content at 96 h (6.6%) and the lowest content at 72 h (4.6%). In contrast, it was observed that the production of fungal proteins is related to the concentration of sugars. Furthermore, fermentation results in a reduction in antinutritional elements, such as heavy metals (As, Cd, Pb, Hg, and Sn), and there is a decrease in ash content during fermentation kinetics. Finally, this work shows that Aspergillus oryzae can assimilate nutrients found in the pretreated Sargassum spp. to produce fungal proteins as a strategy for the food industry.

The effect of charged residue substitutions on the thermodynamics of protein-surface interactions.

The interactions of proteins with surfaces are important in both biological processes and biotechnologies. In contrast to decades of study regarding the biophysics of proteins in bulk solution, however, our mechanistic understanding of the biophysics of proteins interacting with surfaces remains largely qualitative. In response, we have set to explore quantitatively the thermodynamics of protein-surface interactions. In this work, we explore systematically the role of electrostatics in modulating the interaction between proteins and charged surfaces. In particular, we use electrochemistry to explore the extent to which a macroscopic, hydroxyl-coated surface held at a slightly negative potential affects the folding thermodynamics of surface-attached protein variants with different composition of charged amino acids. Doing so, we find that attachment to the surface generally leads to a net stabilization, presumably due to excluded volume effects that reduce the entropy of the unfolded state. The magnitude of this stabilization, however, is strongly correlated with the charged-residue content of the protein. In particular, we find statistically significant correlations with both the net charge of the protein, with greater negative charge leading to less stabilization by the surface, and with the number of arginines, with more arginines leading to greater stabilization. Such findings refine our understanding of protein-surface interactions, providing in turn a guiding rationale to achieve the functional deposition of proteins on artificial surfaces for implementation in, for example, protein-based biotechnologies.

Trichoderma sp. spores and Kluyveromyces marxianus cells magnetic separation: Immobilization on chitosan-coated magnetic nanoparticles.

In the present study, the interactions between chitosan-coated magnetic nanoparticles (C-MNP) and Trichoderma sp. spores as well as Kluyveromyces marxianus cells were studied. By Plackett-Burman design, it was demonstrated that factors which directly influenced on yeast cell immobilization and magnetic separation were inoculum and C-MNP quantity, stirring speed, interaction time, and volume of medium, while in the case of fungal spores, the temperature also was disclosed as an influencing factor. Langmuir and Freundlich models were applied for the mathematical analysis of adsorption isotherms at 30°C. For Trichoderma sp. spore adsorption isotherm, the highest correlation coefficient was observed for lineal function of Langmuir model with a maximum adsorption capacity at 5.00E + 09 spores (C-MNP g-1). Adsorption isotherm of K. marxianus cells was better adjusted to Freundlich model with a constant (Kf) estimated as 2.05E + 08 cells (C-MNP g-1). Both systems may have a novel application in fermentation processes assisted with magnetic separation of biomass.