Differential impacts of furfural and acetic acid on the bioenergetics and fermentation performance of Scheffersomyces stipitis.

Lignocellulosic material is a leading carbon source for economically viable biotechnological processes; however, compounds such furfural and acetic acid exhibit toxicity to yeasts. Nonetheless, research about the molecular mechanism of furfural and acetic acid toxicity is still scarce in yeasts like Scheffersomyces stipitis. Thus, this study aims to elucidate the impact of furfural and acetic acid on S. stipitis regarding bioenergetic and fermentation parameters. Here, we provide evidence that furfural and acetic acid induce a delay in cell growth and extend the lag phase. The mitochondrial membrane potential decreased in all treatments with no significant differences between inhibitors or concentrations. Interestingly, reactive oxygen species increased when the inhibitor concentrations were from 0.1 to 0.3 % (v/v). The glycolytic flux was not significantly (p > 0.05) altered by acetic acid, but furfural caused different effects. Ethanol production decreased significantly (4.32 g·L-1 in furfural and 5.06 g·L-1 in acetic acid) compared to the control (26.3 g·L-1). In contrast, biomass levels were not significantly different in most treatments compared to the control. This study enhances our understanding of the effects of furfural and acetic acid at the mitochondrial level in a pentose-fermenting yeast like S. stipitis.

Properties and Biodegradability of Films Based on Cellulose and Cellulose Nanocrystals from Corn Cob in Mixture with Chitosan.

The increase in consumer demand for more sustainable packaging materials represents an opportunity for biopolymers utilization as an alternative to reduce the environmental impact of plastics. Cellulose (C) and chitosan (CH) are attractive biopolymers for film production due to their high abundance, biodegradability and low toxicity. The objective of this work was to incorporate cellulose nanocrystals (NC) and C extracted from corn cobs in films added with chitosan and to evaluate their properties and biodegradability. The physicochemical (water vapor barrier, moisture content, water solubility and color) and mechanical properties of the films were evaluated. Component interactions using Fourier-transform infrared (FTIR) spectroscopy, surface topography by means of atomic force microscopy (AFM), biodegradability utilizing a fungal mixture and compostability by burying film discs in compost were also determined. The C-NC-CH compared to C-CH films presented a lower moisture content (17.19 ± 1.11% and 20.07 ± 1.01%; w/w, respectively) and water vapor permeability (g m−1 s−1 Pa−1 × 10−12: 1.05 ± 0.15 and 1.57 ± 0.10; w/w, respectively) associated with the NC addition. Significantly high roughness (Rq = 4.90 ± 0.98 nm) was observed in films added to NC, suggesting a decreased homogeneity. The biodegradability test showed larger fungal growth on C-CH films than on CH films (>60% and <10%, respectively) due to the antifungal properties of CH. C extracted from corn cobs resulted in a good option as an alternative packaging material, while the use of NC improved the luminosity and water barrier properties of C-CH films, promoting strong interactions due to hydrogen bonds.

Physicochemical and Antimicrobial Characterization of Beeswax-Starch Food-Grade Nanoemulsions Incorporating Natural Antimicrobials.

Nanoemulsions are feasible delivery systems of lipophilic compounds, showing potential as edible coatings with enhanced functional properties. The aim of this work was to study the effect of emulsifier type (stearic acid (SA), Tween 80 (T80) or Tween 80/Span 60 (T80/S60)) and emulsification process (homogenization, ultrasound or microfluidization) on nanoemulsion formation based on oxidized corn starch, beeswax (BW) and natural antimicrobials (lauric arginate and natamycin). The response variables were physicochemical properties, rheological behavior, wettability and antimicrobial activity of BW-starch nanoemulsions (BW-SN). The BW-SN emulsified using T80 and microfluidized showed the lowest droplet size (77.6 ± 6.2 nm), a polydispersion index of 0.4 ± 0.0 and whiteness index (WI) of 31.8 ± 0.8. This BW-SN exhibited a more negative ζ-potential: -36 ± 4 mV, and Newtonian flow behavior, indicating great stability. BW-SN antimicrobial activity was not affected by microfluidization nor the presence of T80, showing inhibition of the deteriorative fungi R. stolonifer, C. gloeosporioides and B. cinerea, and the pathogenic bacterium S. Saintpaul. In addition, regardless of emulsifier type and emulsification process, BW-SN applied on the tomato surface exhibited low contact angles (38.5° to 48.6°), resulting in efficient wettability (-7.0 mN/m to -8.9 mN/m). These nanoemulsions may be useful to produce edible coatings to preserve fresh-produce quality and safety.

Physical, Structural, Barrier, and Antifungal Characterization of Chitosan-Zein Edible Films with Added Essential Oils.

Edible films (EFs) have gained great interest due to their ability to keep foods safe, maintaining their physical and organoleptic properties for a longer time. The aim of this work was to develop EFs based on a chitosan-zein mixture with three different essential oils (EOs) added: anise, orange, and cinnamon, and to characterize them to establish the relationship between their structural and physical properties. The addition of an EO into an EF significantly affected (p < 0.05) the a* (redness/greenness) and b* (yellowness/blueness) values of the film surface. The EFs presented a refractive index between 1.35 and 1.55, and thus are classified as transparent. The physical properties of EFs with an added EO were improved, and films that incorporated the anise EO showed significantly lower water vapor permeability (1.2 ± 0.1 g mm h-1 m-2 kPa-1) and high hardness (104.3 ± 3.22 MPa). EFs with an added EO were able to inhibit the growth of Penicillium sp. and Rhizopus sp. to a larger extent than without an EO. Films' structural changes were the result of chemical interactions among amino acid side chains from zein, glucosamine from chitosan, and cinnamaldehyde, anethole, or limonene from the EOs as detected by a Raman analysis. The incorporation of an EO in the EFs' formulation could represent an alternative use as coatings to enhance the shelf life of food products.

Electrostatic Fermentation: Molecular Response Insights for Tailored Beer Production.

Electrostatic fermentation avoids the cellular redox imbalance of traditional fermentation, but knowledge gaps exist. This study explores the impact of electrostatic fermentation on the growth, volatile profile, and genetic response of Saccharomyces pastorianus Saflager S-23. The applied voltage (15 and 30 V) in the electrostatic fermentation system increased the growth and substrate utilization of S. pastorianus while decreasing ethanol production. The aromas typically associated with traditional fermentation, such as alcoholic, grape, apple, and sweet notes, were diminished, while aromas like roses, fruits, flowers, and bananas were augmented in electrostatic fermentation. RNA-seq analysis revealed upregulation of genes involved in cell wall structure, oxidoreductase activity, and iron ion binding, while genes associated with protein synthesis, growth control, homeostasis, and membrane function were downregulated under the influence of applied voltage. The electrostatic fermentation system modulates genetic responses and metabolic pathways in yeast, rendering it a promising method for tailored beer production. Demonstrating feasibility under industrial-scale and realistic conditions is crucial for advancing towards commercialization.

Biocontrol Activity of Bacillus altitudinis CH05 and Bacillus tropicus CH13 Isolated from Capsicum annuum L. Seeds against Fungal Strains.

In this study, seed-surface-associated bacteria from fresh fruits of Capsicum spp. were analyzed to explore potential isolates for biocontrol of phytopathogenic fungal strains. A total of 76 bacterial isolates were obtained from three different species of chili pepper (C. annuum L., C. pubescens R. & P., and C. chinense Jacq.), and two isolates were selected via mycelial growth inhibition assays based on their production of volatile organic compounds (VOCs) against six fungal strains. Genomic analysis identified these isolates as Bacillus altitudinis CH05, with a chromosome size of 3,687,823 bp and with 41.25% G+C, and Bacillus tropicus CH13, with a chromosome size of 5,283,706 bp and with 35.24% G+C. Both bacterial strains showed high mycelial growth inhibition capacities against Sclerotium rolfsii, Sclerotinia sp., Rhizoctonia solani, and Alternaria alternata but lower inhibition capacities against Colletotrichum gloesporoides and Fusarium oxysporum. VOC identification was carried out after 24 h of fermentation with 64 VOCs for B. altitudinis CH05 and 53 VOCs for B. tropicus CH13. 2,5-Dimethyl pyrazine and acetoin had the highest relative abundance values in both bacterial strains. Our findings revealed that seed-surface-associated bacteria on Capsicum spp. have the metabolic ability to produce VOCs for biocontrol of fungal strains and have the potential to be used in sustainable agriculture.

PVA-Based Electrospun Biomembranes with Hydrolyzed Collagen and Ethanolic Extract of Hypericum perforatum for Potential Use as Wound Dressing: Fabrication and Characterization.

Biological, physicochemical, structural, and thermal properties of PVA-based electrospun wound dressings added with hydrolyzed collagen (HC) and different concentrations of Hypericum perforatum ethanolic extract (EEHP) were studied. Membrane characterization was carried out by X-ray diffraction, Fourier infrared spectroscopy, differential scanning calorimetry, barrier properties, scanning electron microscopy, image analysis (diameter and pore size), as well as antimicrobial and anti-inflammatory activities. Results showed that the PVA/HC/EEHP materials, fabricated under controlled conditions of temperature and humidity, generated fiber membranes with diameters between 140−390 nm, adequate porosity and pore size for cell growth (67−90% and 4−16 µm, respectively), and good barrier properties (0.005−0.032 g·m−2 s−1) to be used in the treatment of conditions on the skin, and was even better than some commercial products. Finally, they showed to have anti-inflammatory (>80%), and antimicrobial activity against S. aureus and S. epiderm. Furthermore, higher crystalline structure was observed according to the EEHP concentration. In addition, this is the first report in which PVA/HC/EEHP membranes are successfully fabricated and characterized.

Rheological and Antimicrobial Properties of Chitosan and Quinoa Protein Filmogenic Suspensions with Thyme and Rosemary Essential Oils.

Food packaging faces the negative impact of synthetic materials on the environment, and edible coatings offer one alternative from filmogenic suspensions (FS). In this work, an active edible FS based on chitosan (C) and quinoa protein (QP) cross-linked with transglutaminase was produced. Thyme (T) and rosemary (R) essential oils (EOs) were incorporated as antimicrobial agents. Particle size, Z potential, and rheological parameters were evaluated. The antimicrobial activity against Micrococcus luteus (NCIB 8166) and Salmonella sp. (Lignieres 1900) was monitored using atomic force microscopy and image analysis. Results indicate that EOs incorporation into C:QP suspensions did not affect the Z potential, ranging from -46.69 ± 3.19 mV to -46.21 ± 3.83 mV. However, the polydispersity index increased from 0.51 ± 0.07 to 0.80 ± 0.04 in suspensions with EO. The minimum inhibitory concentration of active suspensions against Salmonella sp. was 0.5% (v/v) for thyme and 1% (v/v) for rosemary. Entropy and fractal dimension of the images were used to confirm the antimicrobial effect of EOs, which modified the surface roughness.

Improved Thermal and Reusability Properties of Xylanase by Genipin Cross-Linking to Magnetic Chitosan Particles.

Enzymes are gradually increasingly preferred over chemical processes, but commercial enzyme applications remain limited due to their low stability and low product recovery, so the application of an immobilization technique is required for repeated use. The aims of this work were to produce stable enzyme complexes of cross-linked xylanase on magnetic chitosan, to describe some characteristics of these complexes, and to evaluate the thermal stability of the immobilized enzyme and its reusability. A xylanase was cross-linked to magnetite particles prepared by in situ co-precipitation of iron salts in a chitosan template. The effect of temperature, pH, kinetic parameters, and reusability on free and immobilized xylanase was evaluated. Magnetization, morphology, size, structural change, and thermal behavior of immobilized enzyme were described. 1.0 ± 0.1 µg of xylanase was immobilized per milligram of superparamagnetic chitosan nanoparticles via covalent bonds formed with genipin. Immobilized xylanase showed thermal, pH, and catalytic velocity improvement compared to the free enzyme and can be reused three times. Heterogeneous aggregates of 254 nm were obtained after enzyme immobilization. The immobilization protocol used in this work was successful in retaining enzyme thermal stability and could be important in using natural compounds such as Fe3O4@Chitosan@Xylanase in the harsh temperature condition of relevant industries.