Conjugation of Soybean Proteins 7S/11S Isolate with Glucose/Fructose in Gels through Wet-Heating Maillard Reaction.

Conjugation with glucose (G) and fructose (F) via the Maillard reaction under the wet-heating condition is a natural and non-toxic method of improving the technological functions of 7S/11S proteins in different kinds of gels. It may be used as an affordable supply of emulsifiers and an excellent encapsulating matrix for gels. This study aimed to create a glucose/fructose-conjugated 7S/11S soy protein via the Maillard reaction. The conjugation was confirmed by determining the SDS-PAGE profile and circular dichroism spectra. In addition, these conjugates were comprehensively characterized in terms of grafting degree, browning degree, sulfhydryl content, surface hydrophobicity (H0), and differential scanning calorimetry (DSC) through various reaction times (0, 24, 48, and 72 h) to evaluate their ability to be used in food gels. The functional characteristics of the 7S/11S isolate-G/F conjugate formed at 70 °C, with a high degree of glycosylation and browning, were superior to those obtained at other reaction times. The SDS-PAGE profile indicated that the conjugation between the 7S and 11S proteins and carbohydrate sources of G and F through the Maillard reaction occurred. Secondary structural results revealed that covalent interactions with G and F affected the secondary structural components of 7S/11S proteins, leading to increased random coils. When exposed to moist heating conditions, G and F have significant potential for protein alteration through the Maillard reaction. The results of this study may provide new insights into protein modification and establish the theoretical basis for the therapeutic application of both G and F conjugation with soy proteins in different food matrixes and gels.

Biological synthesis of hybrid silver nanoparticles by Periploca aphylla Dcne. From nanotechnology to biotechnology applications.

Nanotechnology is one of the advanced technologies that have almost universal implications in every field of science. The importance is due to the unique properties of nanoparticles; however, green synthesized nanoparticles are considered eco-friendly. The current project was rationalized to prepare green-synthesized biogenic Periploca aphylla Dcne. silver nanoparticles (Pe-AgNPs) and poly (ethylene glycol) methacrylate coated AgNPs nanocomposites (PEGMA-AgNPs) with higher potential for their application in plant tissue culture for enhancing the biomass of Stevia rebaudiana calli. The increased biomass accumulation (17.61 g/3 plates) was observed on a medium containing virgin Pe-AgNPs 40th days after incubation, while the maximum increase was found by supplementing virgin Pe-AgNPs and PEGMA capped AgNPs (19.56 g/3 plates), compared with control (12.01 g/3 plates). In this study, PEGMA capped AgNPs supplementation also induced the maximum increase in total phenolics content (2.46 mg GAE/g-FW), total flavonoids content (3.68 mg QE/g-FW), SOD activity (53.78 U/ml protein), GSH content (139.75 µg/g FW), antioxidant activity (54.3 mg AAE/g FW), FRAP (54 mg AAE/g FW), and DPPH (76.3%) in S. rebaudiana calli compared with the control. It was concluded that virgin Pe-AgNPs and PEGMA capped AgNPs (hybrid polymer) are potent growth regulator agents and elicitors that can be exploited in the biotechnology field for growth promotion and induction of essential bioactive compounds and secondary metabolites from various commercially important and medicinally valuable plants such as S. rebaudiana without laborious field cultivation.

Molecular mechanism of Cu metal and drought stress resistance triggered by Porostereum spadiceum AGH786 in Solanum lycopersicum L.

Rapid industrialization and global warming have threatened the plants with multiple abiotic stresses, such as heavy metals and drought stress. For crop cultivation, the conventional approach of cleaning the soils by excavation is very costly and not feasible for large scale. Establishing toxin-free and drought-resistant crops is a major challenge in the environment under natural and anthropogenic pressure. In the past decades, copper contamination of agricultural land has become an emerging concern. For dry land reclamation, several new strategies, including bioremediation (phytoremediation and microbial remediation), have been used. Owing to the potential of Cu hyperaccumulators, the current project aims to enhance the drought tolerance and the phytoremediation potential of Solanum lycopersicum L. with the inoculation of copper and 12% polyethylene glycol (PEG)-induced drought stress-tolerant endophytic fungus Porostereum spadiceum AGH786 under the combined stress of copper heavy metal and PEG-induced drought stress. When S. lycopersicum L. was watered with individual stress of copper (Cu) concentration (400 ppm) in the form of copper sulfate (CuSO4.5H2O), 12% PEG-induced drought stress and the combined stress of both negatively affected the growth attributes, hormonal, metabolic, and antioxidant potential, compared with control. However, the multistress-resistant AGH786 endophytic fungus ameliorated the multistress tolerance response in S. lycopersicum L. by positively affecting the growth attributes, hormonal, metabolic, and antioxidant potential, and by restricting the root-to-shoot translocation of Cu and inducing its sequestration in the root tissues of affected plants. AGH786-associated plants exhibited a reduction in the severity of copper (Cu) and drought stress, with higher levels of SlCOPT (Cu transporters) and SlMT (metallothionine) gene expressions in root and shoot tissues, indicating that AGH786 contributed to resistance to copper metal toxicity and drought stress in the host S. lycopersicum L.

Endophytic Fungal Consortia Enhance Basal Drought-Tolerance in Moringa oleifera by Upregulating the Antioxidant Enzyme (APX) through Heat Shock Factors.

Global climate change has imposed harsh environmental conditions such as drought. Naturally, the most compatible fungal consortia operate synergistically to enhance plant growth and ecophysiological responses against abiotic strains. Yet, little is known about the interactions between phytohormone-producing endophytic fungal symbionts and plant growth under drought stress. The existing research was rationalized to recognize the role of newly isolated drought-resistant, antioxidant-rich endophytic fungal consortia hosting a xerophytic plant, Carthamus oxycantha L., inoculated to Moringa oleifera L. grown under drought stress of 8% PEG (polyethylene glycol-8000). Under drought stress, the combined inoculation of endophytic strain Microdochium majus (WA), Meyerozyma guilliermondi (TG), and Aspergillus aculeatus (TL3) exhibited a significant improvement in growth attributes such as shoot fresh weight (1.71-fold), shoot length (0.86-fold), root length (0.65-fold), dry weight (2.18-fold), total chlorophyll (0.46-fold), and carotenoids (0.87-fold) in comparison to control (8% PEG). Primary and secondary metabolites were also increased in M. oleifera inoculated with endophytic consortia, under drought stress, such as proteins (1.3-fold), sugars (0.58-fold), lipids (0.41-fold), phenols (0.36-fold), flavonoids (0.52-fold), proline (0.6-fold), indole acetic acid (IAA) (4.5-fold), gibberellic acid (GA) (0.7-fold), salicylic acid (SA) (0.8-fold), ascorbic acid (ASA) (1.85-fold), while abscisic acid (ABA) level was decreased (-0.61-fold) in comparison to the control (8% PEG). Under drought stress, combined inoculation (WA, TG, TL3) also promoted the antioxidant activities of enzymes such as ascorbate peroxidase (APX) (3.5-fold), catalase (CAT) activity (1.7-fold), and increased the total antioxidant capacity (TAC) (0.78-fold) with reduced reactive oxygen species (ROS) such as H2O2 production (-0.4-fold), compared to control (8% PEG), and stomatal aperture was larger (3.5-fold) with a lesser decrease (-0.02-fold) in water potential. Moreover, combined inoculation (WA, TG, TL3) up regulated the expression of MolHSF3, MolHSF19, and MolAPX genes in M. oleifera under drought stress, compared to the control (8% PEG), is suggestive of an important regulatory role for drought stress tolerance governed by fungal endophytes. The current research supports the exploitation of the compatible endophytic fungi for establishing the tripartite mutualistic symbiosis in M. oleifera to alleviate the adverse effects of drought stress through strong antioxidant activities.

Effect of Chilled Storage on Antioxidant Capacities and Volatile Flavors of Synbiotic Yogurt Made with Probiotic Yeast Saccharomyces boulardii CNCM I-745 in Combination with Inulin.

Fermentation of available sugars in milk by yogurt starter culture initially and later by Saccharomyces boulardii (Probiotic yeast) improves the bioavailability of nutrients and produces bioactive substances and volatile compounds that enhance consumer acceptability. The combination of S. boulardii, a unique species of probiotic yeast, and inulin, an exopolysaccharide used as a prebiotic, showed remarkable probiotic and hydrocolloid properties in dairy products. The present study was designed to study the effect of fermentation and storage on antioxidant and volatile capacities of probiotic and synbiotic yogurt by incorporation of S. boulardii and inulin at 1%, 1.5%, and 2% (w/v), compared with the probiotic and control plain yogurt. All samples were stored at 4 °C, and during these four weeks, they were analyzed in terms of their antioxidant and volatile compounds. The synbiotic yogurt samples having inulin and S. boulardii displayed significantly higher DPPH (2,2-diphenyl-1-picrylhydrazyl) free radical activity values and more values of TPC (total phenol contents) than control plain yogurt. A total of 16 volatile compounds were identified in S5-syn2 and S4-syn1.5, while S3-syn1 and S2-P had 14, compared with the control S1-C plain yogurt samples, which had only 6. The number of volatile compounds increased with the increasing concentration of inulin throughout the storage period. Therefore, this novel synbiotic yogurt with higher antioxidant and volatile compounds, even with chilling storage conditions, will be a good choice for consumer acceptability.

Bio-Molecular analysis of selected food derived Lactiplantibacillus strains for CLA production reveals possibly a complex mechanism.

Lactiplantibacillus is among the most extensively studied bacterial specie belonging to the genus Lactobacillus with proven probiotic and health promoting effects. These beneficial effects are generally strains specific but the underlying molecular mechanisms are still not fully understood. Dissecting the determinants behind probiotic topographies of this bacterium is of particular interest since it would help select strains that stand the best chance of success in clinical trials and potential industrial applications. In the current study, we have compared the oleate hydratase phylogeny of seven selected strains of L. plantarum on the basis of their sequence, physiochemical properties and 3D structures. All the strains were assessed on molecular level visualization to find out the active site residues which take part in binding with linoleic acid (LA) at the time of interactions. From the whole genome sequences, the genes responsible for conjugated linoleic acid (CLA) production were identified by Venn diagrams. Identified genes were further compared phylogenetically by MEGA X and physiochemical parameters were analyzed by utilizing ExPaSy-Protparam. The genes were further analyzed for the secondary structures using PSIPRED and tertiary structure was generated by trRosseta and SWISS-MODEL. For the analysis of molecular interactions, LA was used as a ligand and the docking was performed using AutoDock Vina. The phylogenetic analysis showed a close phylogeny of the strains with publicly available genomes. The best interaction energy with LA was observed as -6.7 kcal/mol. The bacteria perform an important role in the CLA production through LA metabolism. Oleate hydratase genes are involved in the complex mechanism of the saturated conversion of LA in to CLA. The current study provides further insights for CLA production by different strains of L. plantarum. There is an excellent opportunity for future studies to investigate different CLA production mechanisms in different bacterial strains.

Biotransformation of linoleic acid into different metabolites by food derived Lactobacillus plantarum 12-3 and in silico characterization of relevant reactions.

Lactic acid bacteria have been reported to be capable of converting polyunsaturated fatty acids, e.g. linoleic acid (LA) into bioactive and other fatty acid metabolites that are not toxic to the bacteria themselves, but the mechanism of this conversion is not clear. Here we reported for the first time that probiotic L. plantarum 12-3 derived from Tibet kefir when supplemented with LA from 1% to 10% in the MRS medium transformed LA to various fatty acid derivatives. These derivatives formed in the medium were identified with gas chromatography and mass spectrometry. In silico studies were done to confirm the enzymatic reactions responsible for this conversion. We found that L. plantarum 12-3 could convert LA at different concentrations to 8 different fatty acid derivatives. Putative candidate enzymes involved in biotransformation of LA into fatty acid derivatives were identified via whole genome of L. plantarum 12-3, including linoleate isomerase, acetoacetate decarboxylase and dehydrogenase. Therefore, the present study provides further understanding of the mechanism of conversion of LA to health-beneficial fatty acid metabolites in probiotic L. plantarum, which can be explored for potential application in functional foods.

Association study of CCR6 rs3093024 with Rheumatoid Arthritis in a Pakistani cohort.

C-C Chemokine receptor 6 (CCR6), an important protein in inflammatory and immunological responses, has been previously reported to be associated with rheumatoid arthritis (RA). Therefore, in order to replicate these findings, a case-control study was conducted on 500 subjects (including 250 RA patients and 250 healthy controls) of Pakistani origin. The aim of this study was to determine the association of CCR6 rs3093024 variant with RA and identify its role in splicing events using bioinformatics tools. The clinical and demographic characteristics of the patients were collected using a well-designed questionnaire. The genotype frequencies of CCR6 rs3093024 variant were determined using tetra-primer ARMS-PCR (amplification of refractory mutation system-polymerase chain reaction) method. A significant difference was found between CCR6 rs3093024 genotype frequencies [P = 0.0016, χ 2  = 12.915]. Similarly, a significant difference in the allele frequencies between RA patients and healthy controls was also observed [P = 0.0003 and OR (95% CI) = 0.63 (0.49-0.80)]. The stratification of patients showed that there was a significant increase in AA genotype against AG + GG in patients [P = 0.0014, OR (95% CI) = 2.0 (1.32-3.02)]. Furthermore, using bioinformatics analysis, it was found that CCR6 rs3093024 variant might create a potential splicing enhancer motif (SF2/ASF (IgM-BRCA1) with score of 77.92; Threshold 70.53), which might have important impact on the product of this gene. This study suggests that the A variant of CCR6 rs3093024 variant is significantly associated with RA-risk and its G variant is protective in Pakistani population but a multi-cohort large sized population study is needed to elucidate these results. Moreover, functional studies are needed to highlight the effects of this polymorphism on the function of CCR6 gene.

Dose-dependent production of linoleic acid analogues in food derived Lactobacillus plantarum K25 and in silico characterization of relevant reactions.

The objective of this study was to assess and scrutinize the competency of probiotic L. plantarum K25 to produce linoleic acid analogues in the medium supplemented with different concentrations of linoleic acid, ranging from 1% to 10%, in a dose dependent manner. The analogues produced were identified and quantitated by GC-MS and in silico studies were done to confirm enzymatic reactions involved in its conversion. The results showed that L. plantarum K25 could convert linoleic acid at different concentrations to 9 different fatty acid analogues at concentrations ranging from 0.01 to 17.24 mg/L. Among these metabolites, formation of an essential fatty acid, the linolenic acid, in media supplemented with 9% linoleic acid, is being reported for the first time. Putative candidate enzymes involved in biotransformation of linoleic acid into linoleic acid analogues were identified in the whole genome of L. plantarum K25, which was sequenced previously. In silico studies confirmed that many enzymes, including linoleate isomerase and dehydrogenase, may be involved in biotransformation of linoleic acid into linoleic acid analogues. Both enzymes could effectively bind the linoleic acid molecule, mainly by forming hydrogen bonding between the acidic groups of linoleic acid and the proline residues at the active sites of the enzymes, validating putative reaction partners.

In silico characterization of linoleic acid biotransformation to rumenic acid in food derived Lactobacillus plantarum YW11.

Lactobacillus plantarum YW11 capability to convert linoleic acid into conjugated linoleic acid and other metabolites was studied in a dose-dependent manner by supplementing LA at different concentrations. L. plantarum YW11 displayed a uniform distinctive growth curve of CLA and other metabolites at concentrations of LA ranging from 1% (w/v) to 10% (w/v), with slightly increased growth at higher LA concentrations. The biotransformation capability of L. plantarum YW11 evaluated by GC-MS revealed a total of one CLA isomer, i.e. 9-cis,11-trans-octadecadienoic acid, also known as the rumenic acid (RA), one linoleic acid isomer (linoelaidic acid), and LA metabolites: (E)-9-octadecenoic acid ethyl ester, trans, trans-9,12-octadecadienoic acid, propyl ester and stearic acid. All the metabolites of linoleic acid were produced from 1 to 10% LA supplemented MRS media, while surprisingly the only conjugated linoleic acid compound was produced at 10% LA. To assess the presence of putative enzymes, responsible for conversion of LA into CLA, in silico characterization was carried out. The in silico characterization revealed presence of four enzymes (10-linoleic acid hydratase, linoleate isomerase, acetoacetate decarboxylase and dehydrogenase) that may be involved in the production of CLA (rumenic acid) and LA isomers. The biotransformation ability of L. plantarum YW11 to convert LA into RA has great prospects for biotechnological and industrial implications that could be exploited in the future scale-up experiments.

Physicochemical and Microbiological Properties of Synbiotic Yogurt Made with Probiotic Yeast Saccharomyces boulardii in Combination with Inulin.

Saccharomyces boulardii is a unique species of yeast previously characterized as a probiotic strain (CNCM I-745) among a few probiotic yeasts reported to date. Inulin is one of the most common prebiotics that exhibit twisted hydrocolloidal properties in dairy products. The present study was designed to develop a synbiotic yogurt by incorporation of S. boulardii and inulin at 1%, 1.5%, and 2% (w/v), comparing with the probiotic and control plain yogurts. Microrheological, microstructural, microbiological, sensory properties, and volatile compounds of the yogurt samples were evaluated. Microrheological analysis showed that addition of inulin to yogurt slightly reduced the values of G' and G″, while solid-liquid balance (SLB) values confirmed more solid properties of the synbiotic yogurt (0.582~0.595) than the plain yogurt (0.503~0.518). A total of 18 volatile compounds were identified in the synbiotic yogurt, while only five and six compounds were identified in plain and probiotic yogurts, respectively. Physiochemical parameters such as pH, acidity, and protein content were in the normal range (as with the control), while fat content in the synbiotic yogurt decreased significantly. Addition of 1% inulin not only reduced syneresis but also maintained viability of S. boulardii after 28 days of storage. Microstructural and microrheological studies confirmed the dense, compressed, homogeneous structure of the synbiotic yogurt. Thus, addition of inulin improved the textural and sensory properties of the synbiotic yogurt, as well as survival of S. boulardii with viable count above 6.0 log CFU/g in yogurt, as generally required for probiotics. Therefore, novel synbiotic yogurt with desirable quality was developed as an effective carrier for delivery of the probiotic yeast exerting its beneficial health effects.