Search results for: trehalose

Authors: Wessam H. Abd-Elsalam, Mona M. Saber, Samar M. Abouelatta

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Non-steroidal anti-inflammatory drugs (NSAIDs) are considered a double edged sword in inflammatory bowel diseases (IBDs). Some studies reported their advantageous effect in decreasing inflammation, and other studies reported that their use is associated with colitis aggravation. This study aimed to use specifically formulated trehalose-based nano-carriers that targets the colon in an attempt to alleviate inflammation caused by NSAIDs. L-α-phosphatidylcholine (PL), trehalose, and transcutol were used to prepare the trehalosomes (THs), which were also loaded with Tenoxicam(TXM) as a model NSAID. To optimize the formulation variables, a full 23 factorial design, using Design-Expert® software, was performed. The optimized formulation composed of trehalose: PL at a weight ratio of 1:1, 377.72 mg transcutol, and sonicated for 4 min, possessed a spherical shape with a size of 268.61 nm and EE% of 97.83% and released 70.22% of its drug content over 24 h. The superior protective action of TXM loaded THs compared to TXM suspension and drug-free THs was shown by the inhibition of the inflammatory biomarkers, namely; IL-1ß, IL-6, and TNF-alpha levels, as well as oxidative stress markers, measured as GSH and MDA. Improved histopathology of the colonic tissue in male New Zealand rabbits also confirmed the superiority of the TXM loaded THs compared to the unformulated drug or the drug free nano-carriers. Our findings highlight the prosperous role of THs in colon targeting and its anti-inflammatory characteristics in guarding against possible NSAIDs-driven exacerbation of colitis.

Keywords: inflammatory bowel disease, trehalose, trehalosomes, colon targeting

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Authors: S. K. Thind, Aparjot Kaur

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Heat stress is one of the major environmental factors drastically reducing wheat production. Crop heat tolerance can be enhanced by preconditioning of plants by exogenous application of osmoprotectants. Presently, the effect of trehalose pretreatment (at 1 mM, and 1.5 nM) under heat stress of 35±2˚C (moderate) and 40±2˚ (severe) for four and eight hour was conducted in wheat (Tricticum aestivum L.) genotypes viz. HD2967, PBW 175, PBW 343, PBW 621, and PBW 590. Heat stress affects wide spectrum of physiological processes within plants that are irreversibly damaged by stress. Membrane thermal stability (MTS) and cell viability was significantly decreased under heat stress for eight hours. Pretreatment with trehalose improved MTS and cell viability under stress and this effect was more promotory with higher concentration. Thermal stability of photosynthetic apparatus differed markedly between genotypes and Hill reaction activity was recorded more in PBW621 followed by C306 as compared with others. In all genotypes photolysis of water showed decline with increase in temperature stress. Trehalose pretreatment helped in sustaining Hill reaction activity probably by stabilizing the photosynthetic apparatus against heat-induced photo inhibition. Both plant growth and development were affected by temperature in both shoot and root under heat stress. The reduction was compensated partially by trehalose (1.5 mM) application. Adaption to heat stress is associated with the metabolic adjustment which led to accumulation of soluble sugars including non-reducing and reducing for their role in adaptive mechanism. Higher acid invertase activity in shoot of tolerant genotypes appeared to be a characteristic for stress tolerance. As sucrose synthase play central role in sink strength and in studied wheat genotype was positively related to dry matter accumulation. The duration of heat stress for eight hours had more severe effect on these parameters and trehalose application at 1.5 mM ameliorated it to certain extent.

Keywords: heat stress, Triticum aestivum, trehalose, membrane thermal stability, triphenyl tetrazolium chloride, reduction test, growth, sugar metabolism

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Authors: Yun-Chin Chung, Nileema Divate, Gen-Hung Chen, Pei-Ru Huang, Rupesh Divate

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Many environmental factors, such as glucose concentration, ethanol, temperature, osmotic pressure and pH, decrease the production rate of ethanol using yeast as a starter. Fermentation starters with high tolerance to various stresses are always demanded for brewing industry. Trehalose, a storage carbohydrate in cell wall of yeast, plays an important role in tolerance of environmental stress by preserving integrity of plasma membrane and stabilizing proteins. Furan aldehydes are toxic to yeast and the growth rate of yeast is significantly reduced if furan aldehydes were present in the fermentation medium. In yeast, aldehyde reductase is involved in the detoxification of reactive aldehydes and consequently the growth of yeast is improved. The aims of this study were to construct a genetic recombinant Saccharomyces cerevisiae or Pichia pastoris with furfural and HMF degrading and high ethanol tolerance capacities. Yeast strains were engineered by genetic recombination for overexpression of trehalose-6-phosphate synthase gene (tps1) and aldehyde reductase gene (ari1). TPS1 gene was cloned from S. cerevisiae by reverse transcription-polymerase chain reaction (RT-PCR) and then ligated with pGAPZαC vector. The constructed vector, pGAPZC-tps1, was transformed to recombinant yeasts strain with overexpression of ari1. The transformants with pGAPZC-tps1-ari1 were generated called STA (S. cerevisiae) and PTA (P. pastoris) with overexpression of tps1, ari1. PCR with tps1-specific primers and western blot with his-tag confirmed the gene insertion and protein expression of tps1 in the transformants, respectively. The neutral trehalase gene (nth1) of STA was successfully deleted and the novel strain STAΔN will be used for further study, including the measurement of trehalose concentration and ethanol, furfural tolerance assay.

Keywords: genetic recombinant, yeast, ethanol tolerance, trehalase, aldehyde reductase

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Authors: Ankita Pathak, Ashok Munjal, Ravi Parkash

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Plastic responses to multiple environmental stressors in wet or dry seasonal populations of tropical Drosophila species have received less attention. We tested plastic effects of heat hardening, acclimation to drought or starvation; and changes in trehalose, proline and body lipids in D. ananassae flies reared under wet or dry season specific conditions. Wet season flies revealed significant increase in heat knockdown, starvation resistance and body lipids after heat hardening. However, accumulation of proline was observed only after desiccation acclimation of dry season flies while wet season flies elicited no proline but trehalose only. Therefore, drought-induced proline can be a marker metabolite for dry season flies. Further, partial utilization of proline and trehalose under heat hardening reflects their possible thermoprotective effects. Heat hardening elicited cross-protection to starvation stress. Stressor-specific accumulation or utilization, as well as rates of metabolic change for each energy metabolite, were significantly higher in wet season flies than dry season flies. Energy metabolite changes due to inter-related stressors (heat vs. desiccation or starvation) resulted in possible maintenance of energetic homeostasis in wet or dry season flies. Thus, low or high humidity induced plastic changes in energy metabolites can provide cross-protection to seasonally varying climatic stressors.

Keywords: wet-dry seasons, plastic changes, stress related traits, energy metabolites, cross protection

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Authors: Mitsuhiro Hirai, Satoshi Ajito, Nobutaka Shimizu, Noriyuki Igarashi, Hiroki Iwase, Shinichi Takata

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Sugars and polyols are known to be bioprotectants preventing such as protein denaturation and enzyme deactivation and widely used as a nontoxic additive in various industrial and medical products. The mechanism of their protective actions has been explained by specific bindings between biological components and additives, changes in solvent viscosities, and surface tension and free energy changes upon transfer of those components into additive solutions. On the other hand, some organisms having tolerances against extreme environment produce stress proteins and/or accumulate sugars in cells, which is called cryptobiosis. In particular, trehalose has been drawing attention relevant to cryptobiosis under external stress such as high or low temperature, drying, osmotic pressure, and so on. The function of cryptobiosis by trehalose has been explained relevant to the restriction of the intra-and/or-inter-molecular movement by vitrification or from the replacement of water molecule by trehalose. Previous results suggest that the structure and interaction between sugar and water are a key determinant for understanding cryptobiosis. Recently, we have shown direct evidence that the protein hydration (solvation) and structural stability against chemical and thermal denaturation significantly depend on sugar species and glycerol. Sugar and glycerol molecules tend to be preferentially or weakly excluded from the protein surface and preserved the native protein hydration shell. Due to the protective action of the protein hydration shell by those molecules, the protein structure is stabilized against chemical (guanidinium chloride) and thermal denaturation. The protective action depends on sugar species. To understand the above trend and difference in detail, it is essentially important to clarify the characteristics of solutions containing those additives. In this study, by using wide-angle X-ray scattering technique covering a wide spatial region (~3-120 Å), we have clarified structures of sugar solutions with the concentration from 5% w/w to 65% w/w. The sugars measured in the present study were monosaccharides (glucose, fructose, mannose) and disaccharides (sucrose, trehalose, maltose). Due to observed scattering data with a wide spatial resolution, we have succeeded in obtaining information on the internal structure of individual sugar molecules and on the correlation between them. Every sugar gradually shortened the average inter-molecular distance as the concentration increased. The inter-molecular interaction between sugar molecules was essentially showed an exclusive tendency for every sugar, which appeared as the presence of a repulsive correlation hole. This trend was more weakly seen for trehalose compared to other sugars. The intermolecular distance and spread of individual molecule clearly showed the dependence of sugar species. We will discuss the relation between the characteristic of sugar solution and its protective action of biological materials.

Keywords: hydration, protein, sugar, X-ray scattering

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Authors: A. C. Rana, Abhinav Singh Rana

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Layer by layer coating of biocompatible polyelectrolytes converts the liposomes into stable version i.e 'layersomes'. This system was further used to deliver the Amphotericin B through the oral route. Extensive optimization of different process variables resulted in the formation of layersomes with the particle size of 238.4±5.1, PDI of 0.24±0.16, the zeta potential of 34.6±1.3, and entrapment efficiency of 71.3±1.2. TEM analysis further confirmed the formation of spherical particles. Trehalose (10% w/w) resulted in the formation of fluffy and easy to redisperse cake in freeze dried layersomes. Controlled release up to 50 % within 24 h was observed in the case of layersomes. The layersomes were found stable in simulated biological fluids and resulted in the 3.59 fold higher bioavailability in comparison to free Amp-B. Furthermore, the developed formulation was found to be safe in comparison to Fungizone as indicated by blood urea nitrogen (BUN) and creatinine level.

Keywords: amphotericin B, layersomes, liposomes, toxicity

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Authors: Gagan Brar, Munruchi Kaur

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Macrocybe gigantea was collected from the wild, growing as pure white, fleshy, robust fruit bodies in caespitose clusters. Initially, the few ladies collecting these fruiting bodies for cooking revealed their edibility status, which was later confirmed through classical and molecular taxonomy. The culture of this potential wild edible taxa was raised with an aim of domesticating it. Various solid and liquid media were evaluated for their vegetative growth, in which Malt Extract Agar was found to be the best solid medium and Glucose Peptone medium as the best liquid medium. The effect of different temperatures as well as pH was also evaluated for the vegetative growth of M. gigantea, and it was found that it shows maximum vegetative growth at 30° and pH 5. For spawn preparation, various grains viz. Wheat grains, Jowar grains, Bajra grains and Maize grains were evaluated, and it was found that wheat grains boiled for 30 minutes gave the maximum mycelial growth. Mother spawn was thus prepared on wheat grains boiled for 30 minutes. For raising the fruiting bodies, different locally available agro-wastes were tried, and it was found that paddy straw gives the best growth. Both wilds as well as cultivated M. gigantea were compared through HPLC to evaluate the different nutritional and nutraceutical values. For the evaluation of different sugars in wild and cultivated M. gigantea, 15 sugars were taken for analysis. Among these Melezitose, Trehalose, Glucose, Xylose and Mannitol were found in the wild collection of M. gigantea; in the cultivated sample, Melezitose, Trehalose, Xylose and Dulcitol were detected. Among the 20 different amino acids, 18 amino acids were found, except Asparagine and Glutamine in both wild as well as cultivated samples. Among the 37 tested fatty acids, only 6 fatty acids, namely Palmitic acid, Stearic acid, Cis-9 Oleic acid, Linoleic acid, Gamma-Linolenic acid and Tricosanoic acid, were found in both wild and cultivated samples, although the concentration of these fatty acids was more in the cultivated sample. From the various vitamins tested, Vitamin C, D and E were present in both wild and cultivated samples. Both wild as well as cultivated samples were evaluated for the presence of phenols; for this purpose, eleven phenols were taken as standards in HPLC analysis, and it was found that Gallic acid, Resorcinol, Ferulic acid and Pyrogallol were present in the wild mushroom sample whereas in the cultivated sample Ferulic acid, Caffeic Acid, Vanillic acid and Vanillin are present. The flavonoid analysis revealed the presence of Rutin, Naringin and Quercetin in wild M. gigantea, while 5 Naringin, Catechol, Myrecetin, Gossypin and Quercetin were found in cultivated one. From the comparative chromatographic profiling of both wild as well as cultivated M. gigantea, it is concluded that no nutrient loss was found during its cultivation. An increase in percentage of secondary metabolites (i.e., phenols and flavonoids) was found in cultivated one as compared to wild M. gigantea. Thus, from future perspective cultivated species of M. gigantea can be recommended for the commercial purpose as a good food supplement.

Keywords: culture, edible, fruit bodies, wild

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Authors: Pauline Ribert, Gaelle Roudaut, Sebastien Dupont, Laurent Beney

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Yeasts, anhydrobiotic organisms, can survive extreme water disturbances, thanks to the prolonged and reversible suspension of their cellular activity as well as the establishment of a defense arsenal. This property is exploited by many industrialists. One of the protection systems implemented by yeast is the vitrification of its cytoplasm by trehalose. The thermal resistance of dry yeasts is a crucial parameter for their use. However, studies on the thermal resistance of dry yeasts are often based on yeasts produced in laboratory conditions with non-optimal drying processes. We, therefore, propose a study on the thermal resistance of industrial dry yeasts in relation to their thermophysical properties. Heat stress was applied at three temperatures (50, 75, and 100°C) for 10, 30, or 60-minute treatments. The survival of yeasts to these treatments was estimated, and their thermophysical properties were studied by differential scanning calorimetry. The industrial dry yeasts resisted 60 minutes at 50°C and 75°C and 10 minutes at a temperature close to 100°C. At 100°C, yeast was above their glass transition temperature. Industrial dry yeasts are therefore capable of withstanding high thermal stress if maintained in a specific thermophysical state.

Keywords: dry yeast, glass transition, thermal resistance, vitrification

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Authors: Pannaga Pavan Jutur

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Parachlorella kessleri, a marine unicellular green alga belonging to class Trebouxiophyceae, accumulates large amounts of oil, i.e., lipids under nutrient-deprived (-N, -P, and -S) conditions. Understanding their metabolic imprints is important for elucidating the physiological mechanisms of lipid accumulations in this microalga subjected to nutrient deprivation. Metabolic and lipidomic profiles were obtained respectively using gas chromatography-mass spectrometry (GC-MS) of P. kessleri under nutrient starvation (-N, -P and -S) conditions. Relative quantities of more than 100 metabolites were systematically compared in all these three starvation conditions. Our results demonstrate that in lipid metabolism, the quantities of neutral lipids increased significantly followed by the decrease in other metabolites involved in photosynthesis, nitrogen assimilation, etc. In conclusion, the metabolomics and lipidomic profiles have identified a few common metabolites such as citric acid, valine, and trehalose to play a significant role in the overproduction of oil by this microalga subjected to nutrient deprivation. Understanding the entire system through untargeted metabolome profiling will lead to identifying relevant metabolites involved in the biosynthesis and degradation of precursor molecules that may have the potential for biofuel production, aiming towards the vision of tomorrow’s bioenergy needs.

Keywords: algae, biofuels, nutrient stress, omics

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Authors: K. Iu. Hrizhymalska, O. Ol. Andrushkova, I. Iu. Pshenychna

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Aim. To study and copare the side effects of myopia contact correction by soft lenses and orthokeratology lenses among medical students. Patients and methods: 34 students (68 eyes) with moderate and severe myopia, who used contact correction of myopia for 2-4 years, were examined. Some of them used soft lenses, while others - orthokeratology lenses. Methods were used: biomicroscopy of the eye surface, Schirmer's test, Norn's test, survey regarding satisfaction with use. Results. Corneal vascularization along the limbus was noted in 4 (5%) eyes of the examined students. In 8 (11%) eyes, symptoms of mild dry eye disease were detected. 2 (3%) eyes showed signs of meibomitis. Allergic conjunctivitis was observed in 4 (5%) eyes, and a purulent corneal ulcer was present in 1 eye. Surveys have shown that orthokeratology lenses unlike soft lenses don't limit everyday activity (in sports, tourism, swimming etc.), they also don't cause discomfort during temperature changes and reduce existing symptoms of dry eye disease. Conclusion. Thus, myopia contact correction is one of the optimal options among students, which allows to expand physical and mental activity. However, taking into account the frequency of side effects in users of soft contact lenses, it is necessary to carry out prevention and treatment of myopia in medical students, follow the recommendations for use, instill preservative-free tear substitutes with trehalose when symptoms of dry eye appear. Also when side reactions occur, contact correction with soft lenses should be changed to orthokeratology lenses.

Keywords: correction, myopia, soft lenses, orthokeratology, specracles, cornea, dry eye, side effects, refractive errors

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Authors: Lintle Mohase, Ninikoe Lebusa, Mpho Stephen Mafa

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Wheat is an economically important cereal crop. However, the changing climatic conditions that intensify drought in production areas, and additional pest infestation, such as the Russian wheat aphid (RWA, Diuraphis noxia), severely hamper its production. Drought and pest management require an additional water supply through irrigation and applying inorganic nutrients (including silicon) as alternative strategies to mitigate the stress effects. Therefore, other approaches are needed to enhance wheat productivity during drought stress and aphid abundance. Two wheat cultivars were raised under greenhouse conditions, exposed to drought stress, and treated with silicon before infestation with the South African RWA biotype 2 (RWASA2). The morphological evaluations showed that severe drought or a combination of drought and infestation significantly reduced the plant height of wheat cultivars. Silicon treatment did not alleviate the growth reduction. The biochemical responses were measured using spectrophotometric assays with specific substrates. An evaluation of the enzyme activities associated with oxidative stress and defence responses indicated that drought stress increased NADPH oxidase activity, while silicon treatment significantly reduced it in drought-stressed and infested plants. At 48 and 72 hours sampling periods, a combination of silicon, drought and infestation treatment significantly increased peroxidase activity compared to drought and infestation treatment. The treatment also increased β-1,3-glucanase activity 72 hours after infestation. In addition, silicon and drought treatment increased glucose but reduced sucrose accumulation. Furthermore, silicon, drought, and infestation treatment combinations reduced the sucrose content. Finally, silicon significantly increased the trehalose content under severe drought and infestation, evident at 48 and 72-hour sampling periods. Our findings shed light on silicon’s ability to induce protective biochemical responses during drought and aphid infestation.

Keywords: drought, enzyme activity, silicon, soluble sugars, Russian wheat aphid, wheat

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Authors: Chandragouda R. Patil, K. S. Jagadeesh, S. D. Kalolgi

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Biofertilizers containing live microbial cells can mobilize one or more nutrients to plants when applied to either seed or rhizosphere. They form an integral part of nutrient management strategies for sustainable production of agricultural crops. Annually, about 22 tons of lignite-based biofertilizers are being produced and supplied to farmers at the Institute of Organic Farming, University of Agricultural Sciences, Dharwad, Karnataka state India. Although carrier based biofertilizers are common, they have shorter shelf life, poor quality, high contamination, unpredictable field performance and high cost of solid carriers. Hence, liquid formulations are being developed to increase their efficacy and broaden field applicability. An attempt was made to develop liquid formulation of strains of Rhizobium NC-92 (Groundnut), Azospirillum ACD15 both nitrogen-fixing biofertilizers and Pseudomonas striata an efficient P-solubilizing bacteria (PSB). Different concentration of amendments such as additives (glycerol and polyethylene glycol), adjuvants (carboxyl methyl cellulose), gum arabica (GA), surfactant (polysorbate) and trehalose specifically for Azospirillum were found essential. Combinations of formulations of Rhizobium and PSB for groundnut and Azospirillum and PSB for maize were evaluated under field conditions to determine the optimum level of inoculum required. Each biofertilizer strain was inoculated at the rate of 2, 4, 8 ml per kg of seeds and the efficacy of each formulation both individually and in combinations was evaluated against the lignite-based formulation at the rate of 20 g each per kg seeds and a un-inoculated set was included to compare the inoculation effect. The field experiment had 17 treatments in three replicates and the best level of inoculum was decided based on net returns and cost: benefit ratio. In peanut, the combination of 4 ml of Rhizobium and 2 ml of PSB resulted in the highest net returns and higher cost to benefit ratio of 1:2.98 followed by treatment with a combination of 2 ml per kg each of Rhizobium and PSB with a B;C ratio of 1:2.84. The benefits in terms of net returns were to the extent of 16 percent due to inoculation with lignite based formulations while it was up to 48 percent due to the best combination of liquid biofertilizers. In maize combination of liquid formulations consisting of 4 ml of Azospirillum and 2 ml of PSB resulted in the highest net returns; about 53 percent higher than the un-inoculated control and 20 percent higher than the treatment with lignite based formulation. In both the crops inoculation with lignite based formulations significantly increased the net returns over un-inoculated control while levels higher or lesser than 4 ml of Rhizobium and Azospirillum and higher or lesser than 2 ml of PSB were not economical and hence not optimal for these two crops.

Keywords: Rhizobium, Azospirillum, phosphate solubilizing bacteria, liquid formulation, benefit-cost ratio

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Authors: Ting Wu, Feiting Hu, Xinyue Zhang, Shuxin Tang, Xiaoyun Xu

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As a by-product of yuba, processing wastewater of Yuba (PWY) contains many bioactive components such as soybean isoflavones, soybean polysaccharides and soybean oligosaccharides, which is a good source of prebiotics and has a potential of high value utilization. The use of Lactobacillus plantarum to ferment PWY can be considered as a potential biogenic element, which can regulate the balance of intestinal microbiota. In this study, firstly, Lactobacillus plantarum was used to ferment PWY to improve its content of active components and antioxidant activity. Then, the health effect of fermented processing wastewater of yuba (FPWY) was measured in vitro. Finally, microencapsulation technology was used applied to improve the sustained release of FPWY and reduce the loss of active components in the digestion process, as well as to improving the activity of FPWY. The main results are as follows: (1) FPWY presented a good antioxidant capacity with DPPH free radical scavenging ability (0.83 ± 0.01 mmol Trolox/L), ABTS free radical scavenging ability (7.47 ± 0.35 mmol Trolox/L) and iron ion reducing ability (1.11 ± 0.07 mmol Trolox/L). Compared with non-fermented processing wastewater of yuba (NFPWY), there was no significant difference in the content of total soybean isoflavones, but the content of glucoside soybean isoflavones decreased, and aglyconic soybean isoflavones increased significantly. After fermentation, PWY can effectively reduce the soluble monosaccharides, disaccharides and oligosaccharides, such as glucose, fructose, galactose, trehalose, stachyose, maltose, raffinose and sucrose. (2) FPWY can significantly enhance the growth of beneficial bacteria such as Bifidobacterium, Ruminococcus and Akkermansia, significantly inhibit the growth of harmful bacteria E.coli, regulate the structure of intestinal microbiota, and significantly increase the content of short-chain fatty acids such as acetic acid, propionic acid, butyric acid, isovaleric acid. Higher amount of lactic acid in the gut can be further broken down into short chain fatty acids. (3) In order to improve the stability of soybean isoflavones in FPWY during digestion, sodium alginate and chitosan were used as wall materials for embedding. The FPWY freeze-dried powder was embedded by the method of acute-coagulation bath. The results show that when the core wall ratio is 3:1, the concentration of chitosan is 1.5%, the concentration of sodium alginate is 2.0%, and the concentration of calcium is 3%, the embossing rate is 53.20%. In the simulated in vitro digestion stage, the release rate of microcapsules reached 59.36% at the end of gastric digestion and 82.90% at the end of intestinal digestion. Therefore, the core materials with good sustained-release performance of microcapsules were almost all released. The structural analysis results of FPWY microcapsules show that the microcapsules have good mechanical properties. Its hardness, springness, cohesiveness, gumminess, chewiness and resilience were 117.75± 0.21 g, 0.76±0.02, 0.54±0.01, 63.28±0.71 g·sec, 48.03±1.37 g·sec, 0.31±0.01, respectively. Compared with the unembedded FPWY, the infrared spectrum results showed that the microcapsules had embedded effect on the FPWY freeze-dried powder.

Keywords: processing wastewater of yuba, lactobacillus plantarum, intestinal microbiota, microcapsule

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