Innovative Application of Phytochemicals from Fermented Legumes and Spices/Herbs Added in Extruded Snacks

A trend related to adding legume seeds to various products has been observed. This work aimed to use fermented red bean/broad bean seeds and their hulls to produce extruded snacks with more beneficial nutritional properties and good sensory quality. Extruded snacks containing fermented ground seeds (50%) or hull (10%) of red bean/broad bean and corn grits with the addition of selected herbs/spices (0.5%) were prepared. The chemical composition, phenolic profile, antioxidant activity, and sensory quality were analysed. The results showed that the protein content ranged from 9 to 22.9 g 100 g−1, phenolic compounds ranged from 3.97 to 12.80 mg 100 g−1 (with the addition of herbs/spices, even up to 62.88 mg 100 g−1), and antioxidant activities ranged from 4.32 to 10.23 Trolox g−1 (ABTS assay), depending on the type of fermented materials. The addition of ground seeds/hull did not influence the consumer desirability, whereas the addition of selected herbs/spices, particularly lovage, increased it. The application of fermented red bean and broad bean seeds and their hulls, as part of the assumptions of the planetary diet, enabled enrichment of extruded corn products, which are often consumed by vegans and vegetarians, with nutritionally valuable ingredients.

ANTIOXIDANT ACTIVITY OF FOREST MANGGOSTEEN (Garcinia hombroniana Pierre) FRACTION USING DPPH AND ABTS METHOD

This study was carried out to determine the active antioxidant fraction of Garcinia hombroniana bark extract by the DPPH and ABTS method. Along with this, the study also attempts to identify the class of compounds present in the various extract of G. hombroniana by the active fraction. The bark extract of G. hombroniana was prepared by the multilevel maceration method by using three solvents including n-hexane, ethyl acetate, and 96% ethanol. Results of study suggested that n-hexane (HSE), ethyl acetate (EASE) and ethanol 96% extract (ESE) have antioxidant activity with IC50 value of 423 ± 16.72 μg/mL, 284.89 ± 2.7μg/mL, and 10.49 ± 0.19 μg/mL in DPPH assay, while these extracts showed IC50 value of 560.92 ± 48.86 μg/mL, 430.18 ± 16.65 μg/mL, and 13.92 ± 0.57 μg/mL respectively in ABTS assay. Further, fractionated using vacuum column chromatography revealed the presence of five fractions viz., A, B, C, D, and E. Among these, fractions D showed the highest antioxidant activity with an IC50 value of 4.83 ± 0.18 μg/mL and 6.82 ± 0.31 μg/mL in DPPH and ABTS assays. Results of the fractioned analysis and qualitative determination showed that the active fraction of G. hombroniana contained flavonoid, triterpenoid, alkaloid, and saponin compounds, and antioxidant activities of these extracts might be due to the presence of these active ingredients.

Characterization of Two Hydrogen Peroxide Resistant Peroxidases from Rhodococcus opacus 1CP

The dye-decolorizing peroxidases (DyP) are a family of heme-dependent enzymes present on a broad spectrum of microorganisms. While the natural function of these enzymes is not fully understood, their capacity to degrade highly contaminant pigments such as azo dyes or anthraquinones make them excellent candidates for applications in bioremediation and organic synthesis. In this work, two novel DyP peroxidases from the organism Rhodococcus opacus 1CP (DypA and DypB) were cloned and expressed in Escherichia coli. The enzymes were purified and biochemically characterized. The activities of the two DyPs via 2,2′-azino-bis [3-ethylbenzthiazoline-6-sulphonic acid] (ABTS) assay and against Reactive Blue 5 were assessed and optimized. Results showed varying trends for DypA and DypB. Remarkably, these enzymes presented a particularly high tolerance towards H2O2, retaining its activities at about 10 mM H2O2 for DypA and about 4.9 mM H2O2 for DypB.

Pharmacological and Toxicological Appraisal of Promising Enzyme-tyrosinase Isolated from Soil Bacteria-an Update

Aims: The membrane associated tyrosinase is an enzymatically active monomeric glycoprotein which is purified to only a low degree. It has gained importance in the present era due to its antioxidant and immunomodulatory properties as well as applications in industry. Moreover its role in the synthesis of melanin in skin for protection from UV radiations also paved the way towards the better understanding of this enzyme. Study Design: Biochemical and molecular characterization of tyrosinase producing soil bacterium had been followed by the assessment of its antioxidant, cytotoxic and anti-cancer activities. Place and Duration of Study: Whole work had been completed at the Microbiology and Molecular biology labs of IMBB during 2018- 2019. Methodology: Tyrosinase producing species were identified by biochemical characterization followed by their genomic DNA sequencing and BLAST analysis while crude tyrosinase was characterized by Bradford's methods, tyrosinase activity and total protein activity, followed by their molecular characterization on SDS-PAGE. The antioxidant and free radical scavenging properties of tyrosinase were evaluated via DPPH, ABTS and FRAP assays and cell proliferation inhibition and the cytotoxicity was calculated via antitumor and MTT assays. Results: P. putida, B. cereus, B. mycoides, M. luteus, K. pneumonia were found to be tyrosinase producing species while their SDS-PAGE analysis showed that the molecular mass of crude tyrosinase was about 39 kDa. Protein contents, total tyrosinase and specific tyrosinase activity was found to be highest in tyrosinase from B. mycoides [0.008±0.06 mg/ml, 1500±0.06 U/ml and 346820.8±0.03 U/mg of tyrosinase respectively]. The results of biological activities of crude tyrosinase vary from bacteria to bacteria. Tyrosinase from P. putida showed higher antioxidant [66.4±0.01% in DPPH assay, 32.04±0.06%in ABTS assay and 320.6±0.06 in FRAP assay], antitumor [67.8±0.01%] and cytotoxic activity [39±1.0% cell viability], followed by B. Cereus tyrosinase [64.7±0.06% antioxidant power in DPPH assay, 53.41±0.03% in ABTS assay and 159.6±0.06% in FRAP assay, 46.46±0.01% antitumor and 43±0.75% cell viability]. Conclusion: The study revealed that tyrosinase isolated from different bacterial strains depicted optimal percentage of anti-oxidative, anti-proliferative and cellular viability and can be used in the near future for medical and industrial purposes.

Antioxidant Activity in Rheum emodi Wall (Himalayan Rhubarb)

Using natural products as antioxidant agents has been beneficial to replace synthetic products. Efforts have been made to profile the antioxidant capacities of natural resources, such as medicinal plants. The polyphenol content of Himalayan rhubarb, Rheum emodi wall, was measured and the antioxidant activity was determined using DPPH and ABTS+ assay, and the oxidative stress was assessed using SOD enzymatic assay. Five different solvent fractions, n-hexane, n-butanol, ethyl acetate, dichloromethane, and water, were used for screening the antioxidant capacity in effort to determine the optimum extraction solvent. The total phenolic contents for R. emodi fractions ranged from 27.76 to 209.21 mg of gallic acid equivalents (GAE)/g of dry weight. DPPH and ABTS+ assay results are presented into IC50 values, ranged from 21.52 to 2448.79 μg/mL and 90.25 to 1718.05 μg/mL, respectively. The ethyl acetate fraction had the highest antioxidant activity among other fractions. Also, n-butanol and water fractions showed significantly lower IC50 values than the positive control in DPPH radical scavenging activity. The IC50 values of SOD assay of fractions ranged from 2.31 to 64.78 μg/mL. A similar result was observed with ethyl acetate fraction showing the highest SOD radical scavenging activity. The study suggests that the ethyl acetate fraction of R. emodi possess the strongest antioxidant activity, thus the most efficient in extracting antioxidant contents. Moreover, a highly significant correlation was shown between total polyphenol content and antioxidant activity screening assays. The compounds related to the antioxidant activity of R. emodi were identified to myricitrin, myricetin 3-galloyl rhamnoside, and myricetin, which have not been reported in studies about R. emodi before.

Relationships between Structure and Antioxidant Capacity and Activity of Glycosylated Flavonols

The antioxidant capacity (AC) and antioxidant activity (AA) of three flavonols (FLV), aglycones and their glycosylated derivatives were evaluated using 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) assays in various solvents. Findings confirmed that the glycosylation at the 3-position (3-glycosylation) always decreased the AC under most conditions due to substitution of the 3-position hydroxyl group and glycoside disruption in the molecular planarity. The 7-glycosylated derivatives did not have the above effects, thus generally exhibited ACs similar to their aglycones. Glycosylation decreased the AA of kaempferol and isorhamnetin for both assays in methanol, 3-glycosylation inhibited quercetin AA in the ABTS assay. In the DPPH assay, the AA of 3-glycosylated quercetin was significantly higher than quercetin. Using LC–MS/MS analysis, we found that quercetin and quercetin-7-glucoside underwent dimerization during the antioxidant reaction, potentially leading to a decline in AAs. However, 3-glycoside substitution may have hindered dimer formation, thereby allowing the FLVs to retain strong free radical scavenging abilities.

Common Trends and Differences in Antioxidant Activity Analysis of Phenolic Substances Using Single Electron Transfer Based Assays

Numerous assays were developed to measure the antioxidant activity, but each has limitations and the results obtained by different methods are not always comparable. Popular examples are the DPPH and ABTS assay. Our aim was to study similarities and differences of these two assay regarding the measured antioxidant potentials of 24 phenolic compounds using the same measurement and evaluation methods. This should allow conclusions to be drawn as to whether one of the assays is more suitable for measuring specific subgroups like phenolic acids, flavonols, flavanones, dihydrochalcones or flavanols. The assays showed common trends for the mean values of most of the subgroups. Some dihydrochalcones and flavanones did not react with the DPPH radical in contrast to the ABTS radical, leading to significant differences. Therefore, to determine the antioxidant potential of dihydrochalcone or flavanone-rich extracts, the ABTS assay should be preferred. We found that the results of the flavonoids in the DPPH assay were dependent on the Bors criteria, whereas the structure–activity relationship in the ABTS assay was not clear. For the phenolic acids, the results in the ABTS assay were only high for pyrogallol structures, while the DPPH assay was mainly determined by the number of OH groups.

PHYTONUTRIENT SCREENING BY GAS CHROMATOGRAPHY–MASS SPECTROMETRY AND ANTI-AGING PROPERTIES OF GLYCOSMIS PENTAPHYLLA (RETZ.) DC. (SOM-CHUEN) EXTRACTS

Objective: The aims of this study were to screen the phytonutrient constituents and investigate their anti-aging property of leaves and bark ofGlycosmis pentaphylla (Retz.) DC. (GP).Methods: GP is the medicinal plant which used as traditional medicine. Glycosmis pentaphylla leaves (GPL) and Glycosmis pentaphylla bark (GPB) ofGP were extracted with ethanol. The gas chromatography–mass spectrometry was used for phytonutrients analysis. The anti-aging properties wereperformed using ABTS assay, deoxyribose degradation assay, bovine serum albumin (BSA)-fructose model, and matrix metalloproteinase 1 (MMP-1)inhibitory activity.Results: The important phytonutrients of GPL were α-tocopherol, linolenic acid, squalene, stigmasterol, and β-amyrin and for GPB were α-tocopherol,phytol, campesterol, stigmasterol, dictamine, and γ-sitosterol. The percentage inhibition of GPL and GPB extracts at various concentrations wasbetween 16.57–76.05 and 20.66-78.81 by ABTS assay, 68.24–90.06 and 73.83–96.64% by deoxyribose degradation assay, 4.24–99.98 and 54.81–99.94 by BSA-fructose model, and 6.31–81.55 and 1.06–74.45 by MMP-1 inhibitory activity, respectively.Conclusion: Leaves and bark extracts of GP (Retz.) DC. are good sources of important phytonutrients and have the potential in anti-aging property.

Simultaneous Detection and Validation of Analytical Markers of Swertia chirata by HPLC-DAD to Evaluate the Potency of Extracts and Fractions against Antioxidant Potential

The present study is based on the selection of extract and fraction of Swertia chirata plant for the antioxidant potential with HPLC fingerprinting, which includes the simultaneous detection and quantification of four analytical markers protocatechuic acid (PCA), swertiamarin (SM), mangiferin (MF) and amarogentin (AG) by HPLC-DAD. The yield of water extract (SWA), hydroalcoholic extract (SHA) and fractions of hydroalcoholic extracts were evaluated for their antioxidant potential against 2,2-diphenyl-1-picrylhydrazyl-hydrate free radical assay (DPPH assay), 2,2′-azino-bis(3- ethylbenzothiazoline-6-sulfonic acid radical scavenging assay (ABTS assay), total reducing assay (TRA), ferric reducing antioxidant potential assay (FRAP assay), total antioxidant capacity assay (TAC assay). The hydroalcoholic extracts (SHA) can be a better choice as compared to water extract (SWA) due to higher yield of extract (13.680 ± 0.548%) and higher antioxidant activity against DPPH assay, ABTS assay, TRA assay, FRAP assay and TAC assay. In hydroalcoholic extract (SHA), ethyl acetate fraction (SEA) showed most potent activity against DPPH (IC50 = 0.008 ± 0.002 mg/mL) and ABTS (0.025 ± 0.001 mg/mL). n-Hexane fraction of SHA showed higher FRAP (28.664 ± 3.153 μmol/mL) and TAC (3.263 ± 0.325 μmol/mL) value (equivalent to ascorbic acid in μmol/mL) but showed very low yield (0.468 ± 0.018%), SBU showed higher TRA value (0.413 ± 0.309 mg/mL). The ethyl acetate fraction (SEA) can be a choice for antioxidant as it showed second highest FRAP (19.547 ± 2.119 μmol/mL) and TAC (2.750 ± 0.466 μmol/mL) with better yield (2.473 ± 0.594%) as compared to n-hexane (SH) fractions (0.468 ± 0.018%).