Valorization of By-Products from Biofuel Biorefineries: Extraction and Purification of Bioactive Molecules from Post-Fermentation Corn Oil

The aim of this work was to develop innovative and sustainable extraction, concentration, and purification technologies aimed to recover target substances from corn oil, obtained as side stream product of biomass refineries. Residues of bioactive compounds such as carotenoids, phytosterols, tocopherols, and polyphenols could be extracted from this matrix and applied as ingredients for food and feeds, nutraceuticals, pharmaceuticals, and cosmetic products. These molecules are well known for their antioxidant and antiradical capacity, besides other specific biological activities, generically involved in the prevention of chronic and degenerative diseases. The project involved the development of methods for the selective extraction of these minor components, using as suitable extraction technique solid phase extraction. All the extracted and purified fractions were evaluated by NMR spectroscopic analyses and UV–Vis spectrophotometric techniques and characterized by quali-quantitative HPLC analyses. TPC (total phenolic content) and TFC (total flavonoid content) were also determined. DPPH and ABTS radical were used to evaluate radical quenching abilities. Acetylcholinesterase (AChE), amylase, glucosidase, and tyrosinase were selected as enzymes in the enzyme inhibitory assays. The obtained results showed the presence of a complex group of interesting molecules with strong potential in market applications according to circular economy principles.

Microencapsulation of Chokeberry Polyphenols and Volatiles: Application of Alginate and Pectin as Wall Materials

Microencapsulation is a rapidly evolving technology that allows preservation of various high-value, but unstable, compounds, such as polyphenols and volatiles. These components of chokeberry juice are reported to have various health-promoting properties. In the present study, hydrogel beads with alginate or alginate and pectin as wall materials and chokeberry juice as active agent were prepared using Encapsulator B-390. The effects of different compositions of wall material as well as the duration of complexation (30 or 90 min) with hardening solution on microencapsulation of chokeberry polyphenols and volatiles were investigated. Spectrophotometric and HPLC analyses showed that beads with pectin addition contained higher concentrations of polyphenols and anthocyanins compared to those prepared with alginate. Antioxidant activities evaluated with FRAP, CUPRAC, DPPH, and ABTS assays followed the same trend. Encapsulation of volatiles which were determined using GC-MS analysis also depended on the composition of hydrogel beads and in some cases on the time of complexation. Results of this study showed that the selection of the wall material is a relevant factor determining the preservation of polyphenols and volatiles. The incorporation of bioactive compounds in hydrogel beads opens up a wide range of possibilities for the development of functional and innovative foods.

The Contribution of G-Layer Glucose in Salix Clones for Biofuels; Comparative Enzymatic and HPLC Analysis of Stem Cross-Sections

Background Interest on the use of short-rotation willow as a lignocellulose resource for liquid transport fuels has increased greatly over the last ten years. Investigations have shown the advantages and potential of using Salix spp. for such fuels but have also emphasized the wide variations existing in the compositional structure between different species and genotypes in addition to their effects on overall yield. The present work studied the importance of tension wood (TW) as a readily available source of glucose in two-year-old stems of four Salix clones (Tora, Björn, Jorr, Loden). Studies involved application of a novel approach whereby TW-glucose and residual sugars and lignin were quantified using stem cross-sections with results correlated with HPLC analyses of milled wood. Compositional analyses were made for four points along stems and glucose derived from enzyme saccharification of TW gelatinous (G) layers (G-glucose), structural cell wall glucose (CW-glucose) remaining after saccharification and total glucose (T-glucose) determined both theoretically and from HPLC analyses. Comparisons were also made between presence of other characteristic sugars as well as acid-soluble and -insoluble lignin. Results Initial studies showed good agreement between using stem serial sections and milled powder for determining total sugar and lignin. Therefore, sections were used throughout the work. HPLC determination of T-glucose in Salix clones varied between 47.1–52.8%, showing a trend for higher T-glucose with increasing height (Björn, Tora and Jorr). Using histochemical/microscopy and image analysis, Tora (24.2%) and Björn (28.2%) showed greater volumes of % TW than Jorr (15.5%) and Loden (14.0%). Total G-glucose with enzyme saccharification of TW G-layers varied between 3.7–14.7% increasing as the total TW volume increased. CW-glucose measured after enzyme saccharification showed mean values of 41.9–49.1%. Total lignin between and within clones showed small differences with mean variations of 22.4–22.8% before, and 22.4–24.3% after enzyme saccharification. Calculated theoretical and quantified values for CW-glucose at different heights for clones were similar with strong correlation: T-glucose = G-glucose + CW-glucose. Pearson´s correlation displayed a strong and positive correlation between T-glucose and G-glucose, % TW and stem height, and between G-glucose with % TW and stem height. Conclusions The use of stem cross-sections to estimate TW together with enzyme saccharification represents a viable approach for determining freely available G-glucose from TW allowing comparisons between Salix clones. Using stem sections provides for discrete morphological/compositional tissue comparisons between clones with results consistent with traditional wet chemical analysis approaches where entire stems are milled and analyzed. The four clones showed variable TW and presence of total % G-glucose in the order Björn > Tora > Jorr > Loden. Calculated in terms of 1 m3, Salix stems Tora and Björn would contain ca. 0.24 and 0.28 m3 of tension wood representing a significant amount of freely available glucose.

Impact of Simulated Gastrointestinal Conditions on Antiglycoxidant and α-Glucosidase Inhibition Capacities of Cyanidin-3-O-Glucoside

Cyanidin-3-O-glucoside (C3G) is a widespread anthocyanin derivative, which has been reported in vitro to exert potent antioxidant, antiglycation and α-glucosidase inhibition effects. Nevertheless, the physiological relevance of such properties remains uncertain considering its significant instability in gastrointestinal conditions. A simulated digestion procedure was thus instigated to assess the influence of gastric and intestinal media on its chemical integrity and biological activities. HPLC analyses of digested C3G samples confirmed the striking impact of intestinal conditions, as attested by a decomposition ratio of 70%. In contrast, with recovery rates of around 90%, antiglycation, as well as DPPH and ABTS scavenging assays, uniformly revealed a noteworthy persistence of its antiglycoxidant capacities. Remarkably, a prominent increase of its α-glucosidase inhibition activity was even observed after the intestinal phase, suggesting that classical in vitro evaluations might underestimate C3G antidiabetic potential. Consequently, the present data provide novel and specific insights on C3G’s digestive fate, suggesting that the gastrointestinal tract does not profoundly affect its positive action on oxidative and carbonyl stresses. More specifically, it also tends to support its regulating effects on postprandial hyperglycemia and its potential usefulness for diabetes management.

Determination of cytogenetic and epigenetic effects of manganese and copper on Zea mays L.

Heavy metal accumulation and its possible effects are prominent problem for not only human health but also for the environment and plant systems due to that heavy metals are non-biodegradable. In this research, it was aimed to examine the impacts of heavy metals on toxicity and genotoxicity in maize. Seeds of corn were subjected to various concentrations of MnSO4 and CuSO4 for determining their effects on DNA methylation, DNA damage levels, protein and phytohormone alterations. The results revealed that an increase in copper and manganese concentrations causes decrease in soluble protein levels, genomic template stability (GTS) and mitotic index but causes an increase in RAPD profile alterations and DNA hypermethylation. Additionally, HPLC analyses show that CuSO4 and MnSO4 contamination reduces growth-promoting hormones, like gibberellic acid (GA), zeatin (ZA) and indole acetic acid (IAA), and increases the abscisic acid (ABA). This study obviously indicated that CuSO4 and MnSO4 have epigenetic and genotoxic effects. A decrease in the phytohormone level (ZA, GA, and IAA) and an increase in the ABA level under CuSO4 and MnSO4 are thought to be a part of the defense system of maize to struggle with stress.

Production of Minor Ginsenosides C-K and C-Y from Naturally Occurring Major Ginsenosides Using Crude β-Glucosidase Preparation from Submerged Culture of Fomitella fraxinea

Minor ginsenosides, such as compounds (C)-K and C-Y, possess relatively better bioactivity than those of naturally occurring major ginsenosides. Therefore, this study focused on the biotransformation of major ginsenosides into minor ginsenosides using crude β-glucosidase preparation isolated from submerged liquid culture of Fomitella fraxinea (FFEP). FFEP was prepared by ammonium sulfate (30–80%) precipitation from submerged culture of F. fraxinea. FFEP was used to prepare minor ginsenosides from protopanaxadiol (PPD)-type ginsenoside (PPDG-F) or total ginsenoside fraction (TG-F). In addition, biotransformation of major ginsenosides into minor ginsenosides as affected by reaction time and pH were investigated by TLC and HPLC analyses, and the metabolites were also identified by UPLC/negative-ESI-Q-TOF-MS analysis. FFEP biotransformed ginsenosides Rb1 and Rc into C-K via the following pathways: Rd → F2 → C-K for Rb1 and both Rd → F2→ C-K and C-Mc1 → C-Mc → C-K for Rc, respectively, while C-Y is formed from Rb2 via C-O. FFEP can be applied to produce minor ginsenosides C-K and C-Y from PPDG-F or TG-F. To the best of our knowledge, this study is the first to report the production of C-K and C-Y from major ginsenosides by basidiomycete F. fraxinea.