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For health and safety reasons, the search for green, healthy, and low-calorie sweeteners with good taste has become the demand of many consumers. Furthermore, the need for sugar substitutes of natural origin has increased dramatically. In this review, we briefly discussed the safety and health benefits of stevia sweeteners and enumerated some examples of physiological functions of steviol glycosides (SGs), such as anti-inflammatory, anti-obesity, antihypertensive, anti-diabetes, and anticaries, citing various evidence related to their application in the food industry. The latest advances in emerging technologies for extracting and purifying SGs and the process variables and operational strategies were discussed. The impact of the extraction methods and their comparison against the conventional techniques have also been demonstrated. These technologies use minimal energy solvents and simplify subsequent purification stages, making viable alternatives suitable for a possible industrial application. Furthermore, we also elucidated the potential for advancing and applying the natural sweeteners SGs., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

The objective of this work was to develop two freeze-dried snacks from blackcurrant and different sweeteners: honey/isomalt (HI) sweetened with honey/isomalt, and isomalt/stevia (IS) sweetened with isomalt/stevia. Both snacks showed high bioactive compounds retention (>75%) and no significant changes in several physicochemical properties after 6 months storage. Fresh snacks were hard and crunchy, and the perception of consumers within liking categories was: HI: 43%, and IS: 72%. After storage HI snack showed higher acceptance by consumers (75% within liking categories) while IS snack showed a decrease in their acceptance (63% within disliking categories). The penalty analysis showed that the sensory shelf-life of both snacks would be limited by the changes produced in texture and color during storage. HI snack could reach six storage months, while IS would be stored up to 3 months. The use of diverse sweeteners allowed obtaining two different products which could be incorporated into a healthy diet., Competing Interests: DECLARATION OF CONFLICTING INTERESTSThe authors declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.

Steviol glycosides (SGs) are a natural sweetener widely used in the food and beverage industry, but the low solubility and stability of SG aqueous solutions greatly limit their application performance, especially in liquid formulations. In this work, we explore the solubility behavior of rebaudioside A (Reb A) in water, a major component of SGs, with the aim of clarifying the underlying mechanisms of the solubility and stability constraints of SGs, as well as the impact on their multifunctional properties. We demonstrate for the first time that Reb A exhibits hierarchical self-assembly in solutions, forming spherical micelles first when the concentration exceeds its critical micelle concentration (5.071 mg/mL), which then further assemble into large rod-like aggregates. The formation of such large Reb A aggregates is mainly dominated by hydrogen bonding and short-range Coulomb interaction energy, thus leading to the low solubility and precipitation of Reb A solutions. Surprisingly, aggregated Reb A structures display significantly improved organoleptic properties, revealing that self-aggregation can be developed as a simple, efficient, and green strategy for improving the taste profile of SGs. Additionally, the self-aggregation of Reb A at high concentrations impairs active encapsulation and also affects its interfacial and emulsifying properties.

Given the low-calorie, high-sweetness characteristics of steviol glycosides (SGs), developing SGs with improved taste profiles is a key focus. Rebaudioside M8 (Reb M8), a novel non-natural SG derivative obtained through glycosylation at the C-13 position of rebaudioside D (Reb D) using glycosyltransferase UGT94E13, holds promise for further development due to its enhanced sweetness. However, the low catalytic activity of UGT94E13 hampers further research and commercialization. This study aimed to improve the enzymatic activity of UGT94E13 through semirational design, and a variant UGT94E13-F169G/I185G was obtained with the catalytic activity improved by 13.90 times. A cascade reaction involving UGT94E13-F169G/I185G and sucrose synthase At SuSy was established to recycle uridine diphosphate glucose, resulting in an efficient preparation of Reb M8 with a yield of 98%. Moreover, according to the analysis of the distances between the substrate Reb D and enzymes as well as between Reb D and the glucose donor through molecular dynamics simulations, it is found that the positive effect of shortening the distance on glycosylation reaction activity accounts for the improved catalytic activity of UGT94E13-F169G/I185G. Therefore, this study addresses the bottleneck in the efficient production of Reb M8 and provides a foundation for its widespread application in the food industry.

In this study, bacterial growth, postbiotic short-chain fatty acids (SCFAs) formation, and gelation properties of sugar-free probiotic milk gels produced with stevia and inulin as a sugar replacer and synbiotic interactions were investigated with regard to prebiotic/bio-therapeutic potential and consumer preference. Lactobacillus acidophilus and Bifidobacterium animalis subsp. lactis cultures were used in the manufacture of dietetic milk gels. The addition of stevia and inulin promoted the viability of bacteria and enhanced milk gel firmness throughout its shelf life. The activity of the probiotic bacteria was identified to be within the potential prebiotic effects (> 8.30 log 10 cfu mL -1 ) in a food matrix. However, it was determined that especially stevia and stevia + inulin addition increased the survival rate of probiotic bacteria and in vitro total SCFA production with higher scores for consumers' preferences rather than with the addition of stevia alone. Yoghurts containing B. animalis subsp. lactis have improved the instrumental textural properties, whereas yoghurts containing L. acidophilus had higher scores for sensorial attributes., (© 2023. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

Herbal infusions exhibit diverse pharmacological effects, such as antioxidant, anti-inflammatory, anticancer, antihypertensive, and antineurodegenerative activities, which can be attributed to the high content of phenolic compounds ( e.g ., caffeoylquinic acids (CQAs)). In this study, we used ultraperformance liquid chromatography to determine the content of CQAs in the methanolic extracts of model herbs, namely, yerba mate ( Ilex paraguariensis ), stevia ( Stevia rebaudiana ), and Indian camphorweed ( Pluchea indica (L.) Less.). The results revealed that yerba mate had the highest total CQA content (108.05 ± 1.12 mg/g of dry weight). Furthermore, we evaluated the effect of brewing conditions and storage at 4 °C under dark and light conditions on the antioxidant property and total phenolic and CQA contents of a yerba mate infusion. The analysis of the yerba mate infusions prepared with different steeping times, dried leaf weights, and water temperatures revealed that the amount of extracted CQAs was maximized (∼175 mg/150 mL) when 6 g of dried leaves were steeped in hot water for 10 min. A total of 10-day refrigerated storage resulted in no significant changes in the antioxidant activity and total phenolic and CQA contents of an infusion kept in a brown container (dark). However, the antioxidant properties and total phenolic and CQA contents were negatively affected when kept in a clear container, suggesting the detrimental effect of light exposure. Our study provides practical recommendations for improving the preparation and storage of herbal infusions, thus catering to the needs of consumers, food scientists, and commercial producers. Moreover, it is the first study of the influence of light exposure on the content of crucial quality attributes within plant-based beverages., Competing Interests: The authors declare that they have no competing interests., (© 2024 Khaksar et al.)

Stevia rebaudiana Bertoni is a plant native to South America that has gathered much interest in recent decades thanks to diterpene glycosides, called steviosides, which it produces. These compounds are characterised by their sweetness, which is 250-300 times higher than saccharose, and they contain almost no caloric value. Stevia is currently also grown outside the South American continent, in various countries characterised by warm weather. This research aimed to determine whether it is viable to grow Stevia rebaudiana plants in Poland, a country characterised by a cooler climate than the native regions for stevia plants. Additionally, the impact of adding various dosages and forms of nitrogen fertiliser was analysed. It was determined that Stevia rebaudiana grown in Poland is characterised by a rather low concentration of steviosides, although proper nitrogen fertilisation can improve various characteristics of the grown plants. The addition of 100 kg or 150 kg of nitrogen per hectare of the field in the form of urea or ammonium nitrate increased the yield of the stevia plants. The stevioside content can be increased by applying fertilisation using 100 kg or 150 kg of nitrogen per hectare in the form of ammonium sulfate. The total yield of the stevia plants grown in Poland was lower than the yield typically recorded in warmer countries, and the low concentration of steviosides in the plant suggests that more research about growing Stevia rebaudiana in Poland would be needed to develop profitable methods of stevia cultivation.

Background: It is crucial to reduce the high sugar content of fruit yoghurts in response to the excessive weight gain epidemic. The use of alternative sweeteners in yoghurts is often associated with the negative sensory attributes that can have an impact on yoghurt liking. The main objective of this research was to investigate the effect of alternative sweeteners and strawberry puree addition on the temporal sensory profile of yoghurt using multiple-intake temporal check all that apply (TCATA). A novel approach to the statical analysis of the temporal sensory data was employed by using aligned rank transformation-analysis of variance to investigate the differences between sensory attributes within different products and within different intakes., Results: Results showed that the attributes sweet and fruity decreased when the concentration of fruit puree was increased at low concentration of sucrose. Interestingly, when the concentration of fruit puree was increased, fruitiness increased and mouthcoating decreased at low concentration of stevia. With successive intakes, the attributes sweet, sour, creamy and fruity significantly decreased in yoghurts sweetened with sucrose, xylitol and stevia. Yoghurts containing low concentrations of sucrose or xylitol and fruit puree were liked the most. However, stevia-sweetened yoghurts varying in sweetener and puree concentration were not significantly different in liking. In order to investigate the consumer acceptance of yoghurts, a novel approach was used - that is, utilizing TCATA temporal data to investigate temporal drivers of liking for each yoghurt type., Conclusion: The use of multiple statistical analysis to analyse temporal data suggested that both sweetener and puree concentration need to be considered when developing products using alternative sweeteners. © 2023 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry., (© 2023 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.)

Background: Recent studies suggest that some nonnutritive sweeteners (NNS) have deleterious effects on the human gut microbiome (HGM). The effect of steviol glycosides on the HGM has not been well studied., Objective: We aimed to evaluate the effects of stevia- compared with sucrose-sweetened beverages on the HGM and fecal short-chain fatty acid (SCFA) profiles., Methods: Using a randomized, double-blinded, parallel-design study, n = 59 healthy adults [female/male, n = 36/23, aged 31±9 y, body mass index (BMI): 22.6±1.7 kg/m 2 ] consumed 16 oz of a beverage containing either 25% of the acceptable daily intake (ADI) of stevia or 30 g of sucrose daily for 4 weeks followed by a 4-week washout. At weeks 0 (baseline), 4, and 8, the HGM was characterized via shotgun sequencing, fecal SCFA concentrations were measured using ultra-high performance liquid chromatography-tandem mass spectrometry and anthropometric measurements, fasting serum glucose, insulin and lipids, blood pressure, pulse, and 3-d diet records were obtained., Results: There were no significant differences in the HGM or fecal SCFA between the stevia and sucrose groups at baseline (P > 0.05). At week 4 (after intervention), there were no significant differences in the HGM at the phylum, family, genus, or species level between the stevia and sucrose groups and no significant differences in fecal SCFA. At week 4, BMI had increased by 0.3 kg/m 2 (P = 0.013) in sucrose compared with stevia, but all other anthropometric and cardiometabolic measures and food intake did not differ significantly (P > 0.05). At week 8 (after washout), there were no significant differences in the HGM, fecal SFCA, or any anthropometric or cardiometabolic measure between the stevia and sucrose groups (P > 0.05)., Conclusions: Daily consumption of a beverage sweetened with 25% of the ADI of stevia for 4 weeks had no significant effects on the HGM, fecal SCFA, or fasting cardiometabolic measures, compared with daily consumption of a beverage sweetened with 30 g of sucrose., Trial Registration: clinicaltrials.gov as NCT05264636., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

A decoction of the roots (31.6-316 mg/kg) from Stevia serrata Cav. (Asteraceae) as well as the main component (5-150 mg/kg) showed hypoglycemic and antihyperglycemic effects in mice. The fractionation of the active extract led to the isolation of dammaradiene acetate ( 1 ), stevisalioside A ( 2 ), and three new chemical entities characterized by spectroscopic methods and named stevisaliosides B-D ( 3 - 5 ). Glycoside 2 (5 and 50 mg/kg) decreased blood glucose levels and the postprandial peak during oral glucose and insulin tolerance tests in STZ-hyperglycemic mice. Compounds 1 - 5 were tested also against PTP1B 1-400 and showed IC 50 values of 1180.9 ± 0.33, 526.8 ± 0.02, 532.1 ± 0.03, 928.2 ± 0.39, and 31.8 ± 1.09 μM, respectively. Compound 5 showed an IC 50 value comparable to that of ursolic acid (IC 50 = 30.7 ± 0.00 μM). Docking studies revealed that 2 - 5 and their aglycones bind to PTP1B 1-400 in a pocket formed by the C-terminal region. The volatilome of S. serrata was characterized by a high content of ( E )-longipinene, spathulenol, guaiadiene, seychellene, and aromandendrene. Finally, a UHPLC-UV method was developed and validated to quantify the content of 2 in the decoction of the plant.

Stevia rebaudiana Bertoni water extracts have been used as a natural sweetener and customary medicine by the indigenous inhabitants of South America for several hundred years. This plant was sent to Europe in the 16th century and was described by Peter Jacob Esteve in Spain. Recently the food industry has started to employ S. rebaudiana as sweetener using its glycosides after purification. Advertisement claims that Stevia glycosides is good for controling body mass and reducing glycemia. This study's objective was to evaluate the effect of S. rebaudiana leaf extract on Wistar rats as animal model to prove its effectiveness on body mass control, glycemia reduction, and other biochemical parameters. Three groups were randomly formed with 24 males and 24 females: A blank group without any sweetener, a control group drinking water with 10% glucose, and the test group ingesting a 0.94% water extract of S. rebaudiana. Body mass measurements as well as food and drink consumption were daily performed. The experiment lasted 120 days after the specimens were weaned and got used to eating solid food. Euthanasia was done and blood serum was collected to evaluate the following biochemical parameters: Glucose, triglycerides, cholesterol, insulin, glucagon, leptin, ghrelin, and glucose-dependent insulinotropic peptide, GIP. Results indicated that only female rats had statistical differences in body mass gain. No relevant effects either positive or negative were found in the biochemical parameters measured. The crude extracts of S. rebaudiana did not show any relevant changes in biochemical and hormonal profiles, changes nor body mass with respect to the blank and control groups of young and healthy rats in the age range of infancy to youth. According to the results obtained, the therapeutic properties that have been associated to S. rebaudiana consumption especially for body mass control and glycemia reduction, did not occur in young and healthy male and female rats in equivalent age to infants, young children, and youths., Competing Interests: The authors have declared that no competing interest exist., (Copyright: © 2024 Mendoza-Pérez et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Steviol glycosides possess Bola-form amphiphilic structure, which can solubilize hydrophobic phytochemicals and exert physical modification to the hydrophilic matrix. However, the effect of steviol glycosides on the starch hydrogel is still unclear. Herein, the physicochemical properties, in vitro digestibility, and release behavior of starch hydrogel in the presence of steviol glycosides were investigated. The results showed that the addition of steviol glycosides promoted the gelatinization and gelation of starch, and endowed the starch hydrogel with softer texture, larger volume, and higher water holding capacity. The hydrophobic curcumin was well integrated into hydrogel by steviol glycosides, providing the gel with improved colour brilliance. The introduction of steviol glycosides hardly affected the digestibility of starch gel, but it promoted the release rate of curcumin. Notably, this release behavior was pH dependent, which tended to target the alkaline intestine. This work provided some theoretical supports for the development of sugar-free starchy foods., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Bioconversion of Rebaudioside D faces high-cost obstacles. Herein, a novel glycosyltransferase StUGT converting Rebaudioside A to Rebaudioside D was screened and characterized, which exhibits stronger affinity and substrate specificity for Rebaudioside A than previously reported enzymes. A whole-cell catalytic system was thus developed using the StUGT strain. The production of Rebaudioside D was enhanced significantly by enhancing cell permeability, and the maximum production of 6.12 g/L and the highest yield of 98.08% by cell catalyst was obtained by statistical-based optimization. A new cascade process utilizing this recombinant strain and E. coli expressing sucrose synthase was further established to reduce cost through replacing expensive UDPG with sucrose. A StUGT-GsSUS1 system exhibited high catalytic capability, and 5.27 g L -1 Rebaudioside D was achieved finally without UDPG addition by systematic optimization. This is the best performance reported in cell-cascaded biosynthesis, which paves a new cost-effective strategy for sustainable synthesis of scarce premium sweeteners from biomass., (© 2024 Wiley-VCH GmbH.)

Sugar is crucial as an essential nutrient for humans as well as for providing texture, sweetness and so on to food. But with the rise in people's pursuit of health, it is becoming increasingly clear that excessive consumption of sugar can locate a load on the body. It has been that excessive sugar is associated with many diseases, such as dental caries, obesity, diabetes, and coronary heart disease. Therefore, researchers and industries are trying to reduce or substitute sugar in food without affecting the sensory evaluation. Substituting sugar with sweeteners is alternatively becoming the most traditional way to minimize its use. So far, the sweeteners such as stevia and xylitol have been are commercially applied. Several studies have shown that technological innovation can partially compensate for the loss in sweetness as a result of sugar reduction, such as cross-modal interactions that stimulate sweetness with aroma, nanofiltration that filters disaccharides and above, enzyme-catalyzed sugar hydrolysis, and microbial fermentation that turns sugar into sugar alcohol. This review summarizes these studies to enhance the safety and quality of sugar-reduced products, and will provide some theoretical frameworks for the food industry to reduce sugar in foods, meet consumers' needs, and promote human health.

Tartrazine is a yellow colouring agent that is commonly used in foods; however, high dosages of Tartrazine affect fertility and create oxidative stress by generating free radicals. A plant species known as Stevia rebaudiana has natural antioxidants that show promise for protecting testicular tissue. Consequently, this study was intended to examine the ameliorative effect of the aqueous extract of S. rebaudiana (Stevia) on the fertility of male Wistar rats induced by the daily oral intake of Tartrazine. Utilizing gas chromatography-mass spectrometry, phytochemical identification was accomplished for Stevia extract. Study groups were separated into several groups: the first group (the control) got distilled water for up to 56 days; the Stevia group (1000 mg/kg), the Tartrazine group (300 mg/kg) and the Stevia and Tartrazine group (the group was given Tartrazine after 1 h of Stevia extract intake). Also, the oxidative damage in testicular tissues was assessed by measuring the levels of malondialdehyde (MDA) and antioxidants (catalase [CAT], superoxide dismutase [SOD] and glutathione reductase [GSH]). Further, histological alterations were examined. In addition, cyclic AMP-responsive element modulator (Crem) gene expression levels and their relative proteins were measured in the testicular tissues using quantitative real-time polymerase chain reaction and enzyme-linked immunosorbent assays, respectively. Sperm analysis and testosterone concentration were also performed. SPSS version 25 was used for the analysis of results while (p < .05) was regarded as significant. Compared with the control group, the results demonstrated that Tartrazine caused a significant reduction (p < .05) in the testosterone hormone level (0.70 ± 0.21) and the Crem protein quantity (1.21 ± 0.23) in the treated Tartrazine group. Also, it had a significant decrease (p < .05) in sperm motility, viability, count and antioxidant levels. Moreover, there was a significant increase (p < .05) in sperm abnormalities, MDA level (7.40 ± 1.10), kidney and liver function parameters, and DNA degradation in the treated Tartrazine group compared with the control group. On the contrary, the Stevia extract intake enhanced the testosterone (2.50 ± 0.60), antioxidants and Crem protein levels (2.33 ± 0.10) with an improvement in sperm quality in the Stevia and Tartrazine-treated group compared with the Tartrazine group. Stevia also caused a significant decrease (p < .05) in the MDA level (3.20 ± 0.20), and sperm abnormalities with an enhancement of the liver and kidney function parameters in the Stevia and Tartrazine-treated group compared to the Tartrazine group. Stevia administration has a protective effect on the testicular tissues and sperm quality against toxicity induced by Tartrazine exposure, so it will be a good antioxidant drug to be administered daily before daily administration of Tartrazine., (© 2023 John Wiley & Sons Ltd.)

Objective: Diabetes mellitus (DM) has been considered a major problem because of its related complications and growing incidence worldwide. Testicular dysfunction has become a predominant diabetic complication characterized by impaired reproductive function and testicular damage. Stevia rebaudiana Bertoni has been known for its antioxidant effect on diabetes, inflammation, and obesity. The current study investigates the protective effect of Stevia on diabetic-induced testicular injury., Materials and Methods: Sprague Dawley adult male rats were divided into three groups: the control group, the diabetic group, and the diabetic + Stevia group, type 2 diabetes is induced by a high-fat diet (HFD) and a single dose of 35 mg/kg streptozotocin injection. The effects of Stevia were evaluated regarding biochemical, oxidative stress, histopathological and ultrastructural changes, and immunohistochemical expression of vascular endothelial growth factor (VEGF), vascular cell adhesion molecule-1 (VCAM-1), receptor-interacting serine/threonine-protein kinase 1 (RIPK 1), and caspase 3., Results: Stevia extract attenuated the diabetic-induced oxidative stress, restored the testicular architecture, and decreased testicular damage, inflammation, necroptosis, and apoptosis by upregulating VEGF and downregulating VCAM 1, RIPK 1, and caspase 3., Conclusions: The current study highlights the importance of Stevia as an antioxidant anti-inflammatory that ameliorates diabetic-induced testicular injury by modulating oxidative stress, inflammation, necroptosis, and apoptosis.

Inulin is a polymer of d-fructose, characterized by the presence of a terminal glucose, and are a major component of Stevia rebaudiana roots. This type of polymer has nutritional properties and technological applications, such as fat substitutes in low-calorie foods and as the coating of pharmaceuticals. The aim of this study was to evaluate an alternative method for inulin extraction, in terms of extraction time and yield, since the traditional method of extraction under reflux is both time and energy consuming. Using the response surface methodology (RSM) with Box-Behnken design it was observed that the alternative extraction method using autoclave presented similar yields to the reflux-based method, but with a shorter extraction time, 121 °C by 17.41 min 1 H Nuclear Magnetic Resonance and Matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-ToF-MS) analysis showed that inulin crude extract from S. rebaudiana roots obtained by autoclave extraction had a higher degree of polymerization when compared to those obtained by the traditional method. Thus, it is concluded that the proposed method using an autoclave is a faster alternative for the extraction of inulin., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Steviol glycosides obtained from Stevia rebaudiana leaves are increasingly used in the food industry as natural low-calorie sweeteners. Among them, the sweetness of major glycosides composed of glucose residues (e.g., stevioside and rebaudioside A) has been widely studied. However, the properties of minor natural products containing rhamnose or xylose residues are poorly investigated. In this study, five unreported steviol glycosides containing rhamnose or xylose were extracted from our developing stevia leaves, and their sweetness was evaluated. The highly glycosylated steviol glycosides were identified, and their structures were examined by fragmentation analysis using mass spectrometry. Chemical synthesis of these glycosides confirmed their structures and allowed sensory evaluation of minor steviol glycosides. Our study revealed that a xylose-containing glycoside, rebaudioside FX1, exhibits a well-balanced sweetness, and thus, it is a promising candidate for natural sweeteners used in the food industry.

Recently, the use of different herbal products as carbon sources instead of black and green tea in the preparation of traditional kombucha has been investigated. In this study, functional kombucha was prepared by adding Stevia rebaudiana Bertoni leaves, which have special organoleptic properties, to kombucha medium, and some properties of the beverage were analyzed. Tea blends were determined as 100% green tea (control = C), 75% green tea (GT) + 25% Stevia (ST), 50% GT + 50% ST, and 100% ST. On the 15th day of fermentation, gluconic acid (43.12 ± 0.01 g/L) was detected as dominant organic acid in GT75 + ST25 samples compared to group C (p < 0.05). According to physicochemical parameters that determine the drinkability properties of prepared teas, the best results were in GT25 + ST75 compared to group C (p < 0.05). It proved that the highest activity was in GT25 + ST75 on the 10th day in the groups that applied different antioxidant tests (DPPH, MCA, and CUPRAC). The antimicrobial activities of kombucha at 25, 50, 75, and 100% concentrations of GT and ST reached the highest levels in the GT25 + ST75 group in samples after 10 days of fermentation for all selected microorganisms. The results prove that GT25 + ST75 kombucha is a functional product with high drinkability on the 10th day of fermentation and also more beneficial for health due to the phenolic compounds from both green tea and Stevia. Stevia rebaudiana leaves can be suggested that be used as a new substrate and nitrogen source for kombucha production., (© 2023. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

Diabetes mellitus (DM) is a common chronic medical condition characterized by hyperglycemia resulting from abnormal insulin functionality, of which type 2 DM (T2DM) is the predominant form. An inulin-type fructan, denoted as SRRP, was obtained from Stevia rebaudiana roots via hot-water extraction and alcoholic precipitation, which was subsequently purified by column chromatography. The extracted SRRP sample had a molecular weight of 5.4 × 10 3 Da. Structural analyses indicated that SRRP was composed of 2,1-linked-β-D-fructofuranosyl and α-D-glucopyranosyl residues in a ratio of approximately 29 : 1. In vivo assays revealed that SRRP significantly reduced fasting blood glucose levels, improved insulin resistance, decreased oxidative stress, and regulated lipid metabolism in T2DM mouse models. In addition, SRRP altered the diversity of the gut microbiota and its metabolites in T2DM mice; it increased probiotic bacteria and the concentration of short-chain fatty acids and decreased harmful bacteria. The findings demonstrate the potential of SRRP in the treatment of T2DM.

To improve the conversion efficiency of rebaudioside C, this study screened the Paenarthrobacter ilicis CR5301 from soil samples and identified it by 16S rRNA. The conversion experiment proved that P. ilicis CR5301 was capable of converting rebaudioside C. The effects of initial pH, temperature, inoculation amount, and substrate concentration on rebaudioside C conversion rate were investigated. The results showed that the conversion rate of rebaudioside C reached up to 100% when CR5301 was incubated in a conversion medium with an initial pH of 7.0 for 8 h at 28°C and 270 rpm. The conversion time was reduced by at least 16 h compared with previous studies. The conversion product was analyzed and identified as steviol by high performance liquid chromatography, ultra performance liquid chromatography-triple-time of flight mass spectrometer, and Fourier transform infrared spectroscopy methods. In addition, stevioside, rebaudioside A, dulcoside A, and some unknown components in steviol glycosides byproduct were all efficiently converted to steviol. These findings provide an efficient approach to the conversion of rebaudioside C and byproduct to steviol to simplify the subsequent industrial process and improve the reuse value of steviol glycosides., (© The Author(s) 2022. Published by Oxford University Press on behalf of Applied Microbiology International.)

For health and safety concerns, traditional high-calorie sweeteners and artificial sweeteners are gradually replaced in food industries by natural and low-calorie sweeteners. As a natural and high-quality sugar substitute, steviol glycosides (SvGls) are continually scrutinized regarding their safety and application. Recently, the cultivation of organic stevia has been increasing in many parts of Europe and Asia, and it is obvious that there is a vast market for sugar substitutes in the future. Rebaudioside A, the main component of SvGls, is gradually accepted by consumers due to its safe, zero calories, clear, and sweet taste with no significant undesirable characteristics. Hence, it can be used in various foods or dietary supplements as a sweetener. In addition, rebaudioside A has been demonstrated to have many physiological functions, such as antihypertension, anti-diabetes, and anticaries. But so far, there are few comprehensive reviews of rebaudioside A. In this review article, we discuss the physicochemical properties, metabolic process, safety, regulatory, health benefits, and biosynthetic pathway of rebaudioside A and summarize the modification methods and state-of-the-art production and purification techniques of rebaudioside A. Furthermore, the current problems hindering the future production and application of rebaudioside A are analyzed, and suggestions are provided., (© 2022 Institute of Food Technologists®.)

The Nemo's Garden ® project is an alternative production system for areas with scarce cultivable land but significant presence of water; thus, it is an interesting intervention to address the climate crisis. This work aimed to evaluate the micromorphological, biochemical, and phytochemical characteristics of Stevia rebaudiana (Bertoni) Bertoni grown underwater compared to the terrestrial specimens. The micromorphological analyses, performed on the leaves using light microscopy, fluorescence microscopy, and scanning electron microscopy, evidenced a general uniformity of the trichome morphotype and distribution pattern. The histochemical investigation indicated the simultaneous presence of terpenes and polyphenols in the trichome secreted material from the underwater samples and a prevailing polyphenolic content in the terrestrial specimens; this was also confirmed by biochemical analyses (26.6 mg GAE/g DW). The characterization of non-volatile components, performed using HPLC-MS, showed similar chemical profiles in all the samples, which were characterized by phenolic compounds and steviol glycosides. The volatile compounds, evaluated using HS-SPME coupled with GC-MS, showed sesquiterpene hydrocarbons as the main class in all the analyzed samples (80.1-93.9%). However, the control plants were characterized by a higher content of monoterpene hydrocarbons (12.1%). The underwater biosphere environment did not alter S. rebaudiana micro-morphological characters, although slight qualitative changes were evidenced for the compounds produced as a response to the growth conditions.

Steviol glycosides have been widely applied as new sweeteners in food, beverages, health care, and daily chemical industry owing to the properties of high-intensity sweetness, low calorie, and good physiological characteristics. However, most of steviol glycosides have a bitter taste. Their organoleptic properties can be effectively improved by modifying the linked glycosyl units. In this study, UGT94D1, a uridine diphosphate-dependent glycosyltransferase from Sesamum indicum, was reported to selectively glycosylate rebaudioside A (Reb A) for the synthesis of rebaudioside D2 (Reb D2). Furthermore, a cascade reaction system was constructed to synthesize Reb D2 with 94.66% yield by coupling UGT94D1 with sucrose synthase AtSuSy from Arabidopsis thaliana. Thus, our study not only introduced a practical method for the synthesis of steviol glycosides but also provided the possibility for further exploration of Reb D2., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

Stevioside is the major component of Stevia rebaudiana (Bertoni) leaves, family Asteraceae. UPLC-ESI-MS/MS analyses of leaves total methanol extract (TEx) and standardized butanol fraction (BF, 113.5 mg stevioside/g) were performed herein, revealing steviol glycosides, caffeoylquinic acid derivatives, flavonoids, and sesquiterpenoids. Their hepatoprotective activities against radiation-induced toxicity were evaluated compared to pure stevioside. Rats pretreatment with stevioside, TEx, and BF orally for 7 days before subjection to 6.5 Gy whole-body-gamma-irradiation reversed histopathological damages; improved liver functions and restored depleted antioxidants. ALT and reduced-glutathione levels showed insignificant changes, compared to control, by (5.22%, 3.20%, 24.90%) and (-0.47%, -3.95%, -2.63%), respectively. Glutathione- S -transferase, catalase, and MDA levels were significantly ameliorated. Liver tissue molecular profiling showed reduction in elevated TNF-α by 23.83%, 29.06%, 28.34%, respectively, and in NF-kB and COX-2 expression levels via immunohistochemical study. BF showed better hepatoprotective activity than TEx which may be attributed to higher stevioside, flavonoids, and caffeoylquinic acid derivatives content.

Background: Rebaudioside A (RA) and its monoglucosyl derivative, as like rebaudioside D (RD) are the most popular stevia glycosides but possess poor solubility in water, which limited their application as edible surfactants, the applications as in micellar solubilization and drug delivery. Meanwhile, effect of the monoglucosyl attached to RA moiety remains unclear., Results: Monoglucosyl rebaudioside A (RAG1) was synthesized via hydrolyzing the transglycosylation product of RA with 95% of RA converted. RAG1 content in raw reaction mixture was as high as 69.5% of total glycosides, and harvested with a content of 88.2% by simple filtration. The RAG1 exhibited an aqueous solubility of 87 folds of RA or 391 folds of RD at 25 °C. The surface activity of RAG1 solution was higher than RA and invincible to RD. The RAG1 micelles promoted aqueous solubility of idebenone (IDE) up to 500 folds higher at 25 °C. The cumulative release rate of IDE encapsulated in RAG1 micelles was 777.5% or 456.7% higher of that of free IDE in simulated gastric/intestinal fluids in 14 h, respectively. The RAG1-IDE remained the same in 98 days at 25 °C., Conclusion: The α-linked glucosyl to RA induced higher hydrophilicity and surface activity than that resulted by β-linked glucosyl, making RAG1 not only dramatically raise the aqueous solubility of RA, but also endow IDE folds higher in bioaccessibility, yet making the capsule stable at storage. The results would provide a new edible delivery nanocarrier for encapsulation of hydrophobic bioactive components. © 2021 Society of Chemical Industry., (© 2021 Society of Chemical Industry.)

Owing to zero-calorie and advanced organoleptic properties similar to sucrose, the plant-derived rebaudioside M (Reb M) has been considered as a next generation sweetener. However, a low content of Reb M in Stevia rebaudiana Bertoni and low enzymatic activity of UGT76G1, which is an uridine diphosphate glucose (UDPG)-dependent glycosyltransferase with the ability to glycosylate rebaudioside D (Reb D) to produce Reb M through the formation of β-1,3 glycosidic bond, restrict its commercial usage. To improve the catalytic activity of UGT76G1, a variant UGT76G1-T284S/M88L/L200A was obtained by structure-guided evolution, whose catalytic activity toward Reb D increased by 2.38 times compared with UGT76G1-T284S. This allowed us to prepare Reb M on a large-scale with a great yield of 90.50%. Moreover, molecular dynamics simulation illustrated that UGT76G1-T284S/M88L/L200A reduced distances from Reb D to catalytic residues and UDPG. Hence, we report an efficient method for the potential scale production of Reb M in this study.

Stevia rebaudiana Bertoni is a perennial shrub from Paraguay that is nowadays widely cultivated, since it is increasingly being utilized as a sugar substitute in various foodstuffs due to its sweetness and minimal caloric content. These properties of the plant's derivatives have spurred research on their biological activities revealing a multitude of benefits to human health, including antidiabetic, anticariogenic, antioxidant, hypotensive, antihypertensive, antimicrobial, anti-inflammatory and antitumor actions. To our knowledge, no recent reviews have surveyed and reported published work solely on the latter. Consequently, our main objective was to present a concise, literature-based review of the biological actions of stevia derivatives in various tumor types, as studied in in vitro and in vivo models of the disease. With global cancer estimates suggesting a 47% increase in cancer cases by 2040 compared to 2020, the data reviewed in this article should provide a better insight into Stevia rebaudiana and its products as a means of cancer prevention and therapy within the context of a healthy diet.

Background: Stevia rebaudiana is a high value crop due to the strong commercial demand for its metabolites (steviol glycosides) but has limited geographical cultivation range. In non-native environments with different daylength and light quality, Stevia has low germination rates and early flowering resulting in lower biomass and poor yield of the desired metabolites. In this study, artificial lighting with light-emitting diodes (LEDs) was used to determine if different light quality within and outside of the photosynthetically active radiation (PAR) range can be used to improve germination rates and yields for production of steviol glycosides for the herbal supplement and food industry., Results: Plants treated with red and blue light at an intensity of 130 μmol m -2  s -1 supplemented with 5% of UV-A light under a 16-h photoperiod produced the most desirable overall results with a high rate of germination, low percentage of early flowering, and high yields of dry leaf, stevioside and rebaudioside A, 175 days after planting., Conclusion: While red and blue light combinations are effective for plant growth, the use of supplemental non-PAR irradiation of UV-A wavelength significantly and desirably delayed flowering, enhanced germination, biomass, rebaudioside A and stevioside yields, while supplemental green light improved yield of biomass and rebaudioside A, but not stevioside. Overall, the combination of red, blue and UV-A light resulted in the best overall productivity for Stevia rebaudiana. © 2021 Society of Chemical Industry., (© 2021 Society of Chemical Industry.)

Stevia rebaudiana is an important medicinal plant that belongs to the Asteraceae family. The leaves of Stevia rebaudiana are a rich source of many health-promoting agents such as polyphenols, flavonoids, and steviol glycoside, which play a key role in controlling obesity and diabetes. New strategies such as the elicitation of culture media are needed to enhance the productivity of active components. Herein, the Cuscuta reflexa extracts were exploited as elicitors to enhance the productivity of active components. Cuscuta reflexa is one of the parasitic plants that has the ability to elongate very fast and cover the host plant. Consequently, it may be possible that the addition of Cuscuta reflexa extracts to adventitious root cultures (ADR) of Stevia rebaudiana may elongate the root more than control cultures to produce higher quantities of the desired secondary metabolites. Therefore, the main objective of the current study was to investigate the effect of Cuscuta reflexa extract as a biotic elicitor on the biomass accumulation and production of antioxidant secondary metabolite in submerged adventitious root cultures of Stevia rebaudiana . Ten different concentrations of Cuscuta reflexa were added to liquid media containing 0.5 mg/L naphthalene acetic acid (NAA). The growth kinetics of adventitious roots was investigated for a period of 49 days with an interval of 7 days. The maximum biomass accumulation (7.83 g/3 flasks) was observed on medium containing 10 mg/L extract of Cuscuta reflexa on day 49. As the concentration of extract increases in the culture media, the biomass gradually decreases after 49 days of inoculation. In this study, the higher total phenolics content (0.31 mg GAE/g-DW), total flavonoids content (0.22 mg QE/g-DW), and antioxidant activity (85.54%) were observed in 100 mg/L treated cultures. The higher concentration (100 mg/L) of Cuscuta reflexa extract considerably increased the total phenolics content (TPC), total phenolics production (TPP), total flavonoids content (TFC), total flavonoids production (TFP), total polyphenolics content (TPPC), and total polyphenolics production (TPPP). It was concluded that the extract of Cuscuta reflexa moderately improved biomass accumulation but enhanced the synthesis of phenolics, flavonoids, and antioxidant activities. Here, biomass's independent production of secondary metabolites was observed with the addition of extract. The present study will be helpful to scale up adventitious roots culture into a bioreactor for the production of secondary metabolites rather than biomass accumulation in medicinally important Stevia rebaudiana .

Steviol glycosides are the intensely sweet components of extracts from Stevia rebaudiana. These molecules comprise an invariant steviol aglycone decorated with variable glycans and could widely serve as a low-calorie sweetener. However, the most desirable steviol glycosides Reb D and Reb M, devoid of unpleasant aftertaste, are naturally produced only in trace amounts due to low levels of specific β (1-2) glucosylation in Stevia. Here, we report the biochemical and structural characterization of OsUGT91C1, a glycosyltransferase from Oryza sativa, which is efficient at catalyzing β (1-2) glucosylation. The enzyme's ability to bind steviol glycoside substrate in three modes underlies its flexibility to catalyze β (1-2) glucosylation in two distinct orientations as well as β (1-6) glucosylation. Guided by the structural insights, we engineer this enzyme to enhance the desirable β (1-2) glucosylation, eliminate β (1-6) glucosylation, and obtain a promising catalyst for the industrial production of naturally rare but palatable steviol glycosides., (© 2021. The Author(s).)

Background: Stevia leaves were subjected to convective hot-air, infrared and vacuum drying at 40, 60 and 80 °C, followed by an assessment of thermophysical properties and microstructure, along with drying kinetics modelling and evaluation of energy features for all drying operations., Results: Effective moisture diffusivity (D eff ) showed dependency on temperature with values ranging from 1.08 × 10 -12 to 7.43 × 10 -12  m 2  s -1 for convective drying, from 0.71 × 10 -12 to 6.60 × 10 -12  m 2  s -1 for infrared drying, and from 1.29 × 10 -12 to 5.39 × 10 -12  m 2  s -1 for vacuum drying. The thermal properties of the dried Stevia leaves under different drying conditions showed values of density, specific heat, thermal diffusivity, thermal conductivity and thermal effusivity ranging from 95.6 to 116.2 kg m -3 , 3050 to 3900 J kg -1  K -1 , 4.28 × 10 -7 to 5.60 × 10 -7  m 2  s -1 , 0.16 to 0.23 W m -1  K -1 and 244 to 305 W s 0.5  m -2  K -1 , respectively. As for microstructure, convective hot-air drying showed better preserved leaf characteristics, compared to infrared- and vacuum-drying, whereby scanning electron microscopy (SEM) image analysis also revealed noticeable differences at higher temperatures. Statistical analysis showed that the Midilli-Kuçuk model fitted best the experimental data of drying curves (0.961 < r 2  < 0.999, 0.000064 < SSE < 0.005359, and 0.000074 < χ 2  < 0.006278). Comparison of the drying methods with respect to energy features showed that convective drying at 80 °C led to lowest specific energy consumption (61.86 kW h kg -1 ) with highest efficiency (8.5%)., Conclusion: The results of this study contribute to a better understanding of the drying behaviour and showed that thermophysical properties of dried Stevia leaves and energy features are affected by drying methods. © 2021 Society of Chemical Industry., (© 2021 Society of Chemical Industry.)

Steviol glycosides were subjected to bacteria present in a soil sample collected from a Stevia plantation in Paraguay. During the incubation experiments, next to the aglycon steviol, steviol degradation products were also formed. X-ray analysis and NMR methods in combination with chemical synthesis and GIAO NMR calculations were used to fully characterize the structure of these compounds as a tricyclic ketone and the corresponding reduced form. They were nicknamed monicanone and monicanol . The latter has the ( S )-configuration at the alcohol site.

Obesity is a public health problem characterized by increased body weight due to abnormal adipose tissue expansion. Bioactive compound consumption from the diet or intake of dietary supplements is one of the possible ways to control obesity. Natural products with adipogenesis-regulating potential act as obesity treatments. We evaluated the synergistic antiangiogenesis, antiadipogenic and antilipogenic efficacy of standardized rebaudioside A, sativoside, and theasaponin E1 formulations (RASE1) in vitro in human umbilical vein endothelial cells (HUVECs), 3T3-L1 preadipocytes respectively, and in vivo using a high-fat and carbohydrate diet-induced obesity mouse model. Orlistat was used as a positive control, while untreated cells and animals were normal controls (NCs). Adipose tissue, liver, and blood were analyzed after dissection. Extracted stevia compounds and green tea seed saponin E1 exhibited pronounced antiobesity effects when combined. RASE1 inhibited HUVEC proliferation and tube formation by suppressing VEGFR2, NF- κ B, PIK3, and-catenin beta-1 expression levels. RASE1 inhibited 3T3-L1 adipocyte differentiation and lipid accumulation by downregulating adipogenesis- and lipogenesis-promoting genes. RASE1 oral administration reduced mouse body and body fat pad weight and blood cholesterol, TG, ALT, AST, glucose, insulin, and adipokine levels. RASE1 suppressed adipogenic and lipid metabolism gene expression in mouse adipose and liver tissues and enhanced AMP-activated protein kinase levels in liver and adipose tissues and in serum adiponectin. RASE1 suppressed the NF- κ B pathway and proinflammatory cytokines IL-10, IL-6, and TNF- α levels in mice which involve inflammation and progression of obesity. The overall results indicate RASE1 is a potential therapeutic formulation and functional food for treating or preventing obesity and inflammation., Competing Interests: The authors declare that they have no conflict of interest., (Copyright © 2021 Muhammad Imran Khan et al.)

Stevia rebaudiana Bertoni is a native plant to Paraguay. The extracts have been used as a famous sweetening agent, and the bioactive components derived from stevia possess a broad spectrum of therapeutical potential for various illnesses. Among its medicinal benefits are anti-hypertensive, anti-tumorigenic, anti-diabetic, and anti-hyperlipidemia. Statins (3-hydro-3-methylglutaryl-coenzyme A reductase inhibitor) are a class of drugs used to treat atherosclerosis. Statins are explicitly targeting the HMG-CoA reductase, an enzyme in the rate-limiting step of cholesterol biosynthesis. Despite being widely used in regulating plasma cholesterol levels, the adverse effects of the drug are a significant concern among clinicians and patients. Hence, steviol glycosides derived from stevia have been proposed as an alternative in replacing statins. Diterpene glycosides from stevia, such as stevioside and rebaudioside A have been evaluated for their efficacy in alleviating cholesterol levels. These glycosides are a potential candidate in treating and preventing atherosclerosis provoked by circulating lipid retention in the sub-endothelial lining of the artery. The present review is an effort to integrate the pathogenesis of atherosclerosis, involvement of lipid droplets biogenesis and its associated proteins in atherogenesis, current approaches to treat atherosclerosis, and pharmacological potential of stevia in treating the disease., (Copyright © 2021 The Authors. Published by Elsevier Masson SAS.. All rights reserved.)

Artificial neural network (ANN), as one of the artificial intelligence methods, has been widely using in HPLC studies for modelling purposes. Stevia rebaudiana is an important industrial plant due to its bio-sweetener molecule, rebaudioside-a, in its leaves. Although rebaudioside-a is up to 300-fold sweeter than sucrose, its calorie is almost zero. In this study, HPLC optimization of rebaudioside-a was studied and the optimization data based on multilinear gradient retention times were modelled by ANN. The input parameters were selected as concentrations, column temperatures, initial acetonitrile percentage for the first step of gradient elution, initial acetonitrile percentage for the second step of gradient elution, slope of acetonitrile, wavelengths, flow rates. The retention time was the output. Also, dried S. rebaudiana leaves were extracted and the concentrations were evaluated by HPLC. According to the ANN results, the most effective parameters on the prediction of non-linear gradient retention time for rebaudioside-a were found as flow rate and initial acetonitrile percentage for the second step of gradient. The best back propagation was selected as Levenberg-Marquardt algorithm. The highest rebaudioside-a level was found as 96.53 ± 6.36 μg mL -1 . ANN modelling methods can be used in preparative HPLC applications to estimate the retention time of steviol glycosides., (Copyright © 2021 Elsevier Inc. All rights reserved.)

The aim of this research was to investigate and compare the antioxidant, anti-tyrosinase, anti-aging, and anti-inflammatory activities of 16 herbal extracts for topical application in cosmetic/cosmeceutical products. Herbal plant materials were extracted by infusion in boiled water for 15 min. The total phenolic content and total flavonoid content of each extract were investigated by the Folin-Ciocalteu and aluminum chloride methods, respectively. Antioxidant activities were investigated using 2,2'-diphenyl-1-picrylhydrazyl and a ferric reducing antioxidant power assay. Anti-tyrosinase and anti-aging activities were investigated using an in vitro enzymatic-spectrophotometric method. Anti-inflammatory activities were investigated using an enzyme-linked immunosorbent assay. The findings show that the Stevia rebaudiana extract has the most significant levels of both phenols and flavonoids (p<0.05). The S. rebaudiana, Rosa damascene, and Phyllanthus emblica extracts possessed the most significant antioxidant activities (p<0.05) and a promising whitening effect with moderate anti-tyrosinase activities. Furthermore, the Echinacea purpurea extract possessed the most significant anti-collagenase (78.5±0.0 %), anti-elastase (69.0±1.4 %), and anti-hyaluronidase activity (64.2±0.3 %). The Morus alba extract possessed the most significant anti-inflammatory activity since it could inhibit the secretion of interleukin-6 and tumor necrosis factor-α (p<0.05). Therefore, these herbal extracts have promising skin benefits and have potential for use as active ingredients in cosmetic/cosmeceutical products., (© 2021 Wiley-VHCA AG, Zurich, Switzerland.)

Steviol glycosides are well-known food sweeteners; their consumption has steadily increased over time. A pretreatment method was developed and validated to better separate rebaudioside A and stevioside from various protein-rich and fatty foods for quantification. This method was applied to soy sauce in liquid type and fish cake and coffee in solid type. Parameters such as linearity, limit of detection (LOD), limit of quantification (LOQ), accuracy, and precision were calculated. Calibration curves were linear in the working range of 5-100 mg/l, with coefficients of determination ≥0.99. The LOD and LOQ were in the ranges of 0.16-0.39 and 0.52-1.28 mg/kg, respectively. The percentage recoveries of the fortified samples were in the 88.01%-103.09% range, and the relative standard deviation was <10%. Method validation predicted a desirable accuracy, linearity, and precision. Therefore, the developed method can be practically applied for the quantitation of steviol glycosides in various foods, including soy sauce in liquid type and fish cake and coffee in solid type., (© 2021 Institute of Food Technologists®.)

Skyr yogurts have been gaining prominence because of their different sensory characteristics. Due to their healthy appeal, the use of natural sweeteners to replace sucrose in this type of yogurt can be an alternative for incorporating a sweet taste, in addition to increasing the functionality of the product through the incorporation of prebiotics. This study aimed to determine whether the addition of fructooligosaccharide (FOS), sucrose, stevia, and thaumatin affects the sensory profile of the skyr yogurt with mango pulp and its acceptance in two Brazilian regions. Eight formulations of skyr with mango pulp were developed. The compositional parameters evaluated were moisture, protein, lipids, ash, and carbohydrate. The tests performed were ideal sweetness and mango flavor, sweetness equivalence for each sweetener used, Quantitative Descriptive Analysis (QDA), and consumer testing in the Southeast and Northeast regions of Brazil. In general, the addition of FOS did not impact the characteristics of the formulated skyr yogurt. The type of sweetener had an impact on the sensory profile and acceptance of the skyr yogurt, affected characteristics such as mango flavor, sweet taste, sweet aftertaste, bitter taste, bitter aftertaste, and metallic flavor. The results of the affective test demonstrated that, for consumers in the Southeast, mango flavor is a positive attribute in this yogurt, and for Northeastern consumers, in addition to mango flavor, sweetness must also be taken into consideration. PRACTICAL APPLICATION: This study may be useful for the dairy industry because in the literature, there is still a lack of sensory studies of skyr yogurt, especially when sucrose substitutes are used. The results of the consumer test in this work reinforce the importance of studies related to consumer preferences with cultural differences., (© 2021 Institute of Food Technologists®.)

Nonnutritive sweeteners (NNSs) are widely employed as dietary substitutes for classical sugars thanks to their safety profile and low toxicity. In this study, a re-evaluation of the biological effects of steviol (1), the main metabolite from Stevia rebaudiana glycosides, was performed using the Inverse Virtual Screening (IVS) target fishing computational approach. Starting from well-known pharmacological properties of Stevia rebaudiana glycosides, this computational tool was employed for predicting the putative interacting targets of 1 and, afterwards, of its five synthetic ester derivatives 2-6, accounting a large panel of proteins involved in cancer and inflammation events. Applying this methodology, the farnesoid X receptor (FXR) was identified as the putative target partner of 1-6. The predicted ligand-protein interactions were corroborated by transactivation assays, specifically disclosing the agonistic activity of 1 and the antagonistic activities of 2-6 on FXR. The reported results highlight the feasibility of IVS as a fast and potent tool for predicting the interacting targets of query compounds, addressing the re-evaluation of their bioactivity. In light of the obtained results, the presumably safe profile of known compounds, such as the case of steviol (1), is critically discussed., (Copyright © 2021 Elsevier Inc. All rights reserved.)

Mother liquor sugar (MLS), as the by-product of stevia production, contained ~65% steviol glycosides (SGs). Recovery of the SGs from MLS was achieved by crystallization coupled with reversed-phase chromatography. Crystallization was done by dissolving MLS in methanol solution and SGs were crystallized due to the polarity difference from the medium. Composition of SGs crystals differed with various temperature, time, solid-to-liquid ratio and water content. SGs were 42.25% recovered with high purity of 96.89% under optimal conditions (solid-to-liquid ratio = 1:5 (w/v), T = 25 °C, t = 24 h) in absolute methanol. The liquid phase after crystallization was subsequently subjected to reversed-phase chromatography, whereby the impurities were firstly eluted with 35% (v/v) ethanol solution and the purified SGs were then desorbed by absolute ethanol, finally recovering 95.20% of SGs in the purity of 98.08%. The total SGs recovery of the whole procedure was 97.23%. The two-step purification was easy-to-operate and feasible to scale-up for industrial application., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

The Stevia genus (Asteraceae) comprises around 230 species, distributed from the southern United States to the South American Andean region. Stevia rebaudiana , a Paraguayan herb that produces an intensely sweet diterpene glycoside called stevioside, is the most relevant member of this genus. Apart from S. rebaudiana , many other species belonging to the Stevia genus are considered medicinal and have been popularly used to treat different ailments. The members from this genus produce sesquiterpene lactones, diterpenes, longipinanes, and flavonoids as the main types of phytochemicals. Many pharmacological activities have been described for Stevia extracts and isolated compounds, antioxidant, antiparasitic, antiviral, anti-inflammatory, and antiproliferative activities being the most frequently mentioned. This review aims to present an update of the Stevia genus covering ethnobotanical aspects and traditional uses, phytochemistry, and biological activities of the extracts and isolated compounds.

The partial or total hydrolysis of (3 R ,4 S ,5 S ,6 S ,9 R ,10 R ,11 R )-9,13-diangeloyloxylongipinan-1-one ( 1 ), isolated from the roots of Stevia viscida , gave alcohols 2 or 3 , respectively, which were subjected to molecular rearrangements with boron trifluoride etherate. Compound 2 afforded (3 R ,4 R ,5 R ,6 S ,9 R ,10 S ,11 S )-11,13-oxyneomorelian-1-one ( 10 ) and (4 S ,5 R ,6 S ,8 S ,10 R )-10,13-oxyneojiquilp-2-en-1-one ( 11 ), both possessing novel sesquiterpenoid skeletons. In turn, 3 provided (3 R ,4 R ,5 S ,6 S ,9 R ,11 R )-13-hydroxymoreli-10(14)-en-1-one ( 7 ) and 10 . Acetylation of 3 gave 4 , thus allowing reduction of the C-1 carbonyl group to yield 5 , which was rearranged to (1 S ,3 R ,4 S ,5 S ,6 S ,9 R ,10 R ,11 R )-13-acetoxy-9,11-epoxyjiquilpane ( 6 ), while an attempt to mesylate 3 directly gave rearranged (3 R ,4 R ,5 S ,6 S ,9 R ,11 R )-13-mesyloxymoreli-10(14)-en-1-one ( 8 ) through expulsion of the C-9 mesylate group by the antiperiplanar C-4-C-10 bond migration to C-4-C-9. In addition, treatment of 1 with boron trifluoride etherate generated (3 R ,4 R ,5 S ,6 S ,9 R ,11 R )-13-angeloyloxymoreli-10(14)-en-1-one ( 9 ). The structures of 2 - 11 were elucidated by 1D and 2D NMR experiments and those of 2 , 3 , 8 , 10 , and 11 were confirmed by single-crystal X-ray diffraction analysis.

As currently defined, the exposome represents the lifetime exposure measure of an individual to all potential external genetic influences and their impact on health. Although intentionally added chemicals (e.g., food additives) and food contact materials (e.g., packaging, pesticides) have been assessed for safety to some degree, the full extent to which they can affect health and reproduction has not been reported. The aim of this study was to determine the in vitro and in vivo effects of food additives on the male rat brain and sperm/testes, particularly through oxidative stress. Results from our in vitro study demonstrated that the administration of the common food additive, stevioside, a major component of the common sweetener stevia, as well as the preservatives, diphenyl and orthophenyl phenol (OPP), induced reactive oxygen species (ROS) production in sperm, and led to sperm dysfunction. These effects were inhibited by the addition of the antioxidant α-tocopherol. Moreover, OPP treatment (1/10,000 of no observed adverse effect) induced ROS production in sperm and lipid peroxidation in the epididymis and hippocampus after two weeks in vivo. Furthermore, 4-hydroxynonenal-positive cells, indicating ROS-generated protein modifications, were detected in spermatocytes in the testes and granular cell layer of the dentate gyrus in the brain. Treatment with α-tocopherol significantly improved oxidative stress. Our study suggests that certain food additives may affect sperm function and induce oxidative stress in the testes and brain, resulting in infertility and short-term memory loss, and some antioxidants may improve these dysfunctions., (Copyright © 2021 Elsevier Inc. All rights reserved.)

Stevia rebaudiana (Bertoni) Bertoni is a plant species native to Brazil and Paraguay well-known by the sweet taste of their leaves. Since the recognition of rebaudioside A and other steviol glycosides as generally recognized as safe by the United States Food and Drug Administration in 2008 and grant of marketing approval by the European Union in 2011, the species has been widely cultivated and studied in several countries. Several efforts have been dedicated to the isolation and structure elucidation of minor components searching for novel non-caloric sugar substitutes with improved organoleptic properties. The present review provides an overview of the main chemical approaches found in the literature for identification and structural differentiation of diterpene glycosides from Stevia rebaudiana : High-performance Thin-Layer Chromatography, High-Performance Liquid Chromatography, Electrospray Ionization Mass Spectrometry and Nuclear Magnetic Resonance Spectroscopy. Modification of diterpene glycosides by chemical and enzymatic reactions together with some strategies to scale up of the purification process saving costs are also discussed. A list of natural diterpene glycosides, some examples of chemically modified and of enzymatically modified diterpene glycosides reported from 1931 to February 2021 were compiled using the scientific databases Google Scholar, ScienceDirect and PubMed.

Ethnopharmacological Relevance: Stevia rebaudiana Bertoni is a perennial herb that belongs to the Asteraceae family. It is a natural sweetener plant known as "Sweet Leaf", "Sweet Herbs" and "Honey Leaf", which is estimated to be 300 times more sweetening than sugar cane. Stevia has been used as a traditional treatment for diabetes in many countries for hundreds of years. Several animal studies referred to the antihyperglycemic activity of stevia. However, the combined use of stevia with saxagliptin has not been studied so far, so this study has been done. The aim of the present study was to evaluate the antihyperglycemic effect of stevia alone and in combination with saxagliptin., Materials and Methods: Diabetes was induced in rats by i.p. injection of streptozotocin and nicotinamide. Animals were divided into five groups, each contains eight rats. Group I: included negative controland group II: included diabetic control that received saline. Group III: included diabetic rats that received 400 mg/kg/day stevia aqueous extract. Group IV: included diabetic rats that received saxagliptin 10 mg/kg/day. Group V: included diabetic rats that received stevia 400 mg/kg + saxagliptin 10 mg/kg. Food and water intake were measured daily while body weight was measured weekly. After 3 weeks animals were sacrificed and blood and tissue samples were collected. Fasting blood glucose (FBG), serum insulin, serum dipeptidylepeptidase-4 (DPP-4), TC, TGs, LDL, HDL, GSH and MDA were measured in treated and control rats by colorimetric and ELISA methods., Results: Both stevia and saxagliptin significantly reduced food, water intake, body weight and FBG. Stevia with saxagliptin produced more significant decrease in FBG. While serum insulin increased significantly in stevia, saxagliptin treated groups and their combination. Serum DPP-4 decreased significantly in all treated groups, concerning lipid profile, stevia and saxagliptin notably lowered TC, TGs, and LDL and increased HDL. Both stevia and saxagliptin remarkably decreased MDA and increased GSH compared to diabetic rats. In addition, stevia significantly improved the antidiabetic effects of saxagliptin., Conclusion: Stevia has an antihyperglycemic effect and could enhance the antidiabetic activity of saxagliptin. DPP-4 attenuation, antihyperlipidemic and antioxidant activity as well as improvement of insulin sensitivity may be involved in the antidiabetic action of stevia., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Background and Objective: Contraceptive pills are chemical substances used as a means to prevent pregnancy, but they have several effects, including high lipid profile and in many cases, patients with heart and blood diseases cannot use it as a contraceptive helps in increasing the risk of cardiovascular diseases (CVD). A Stevia extract with high sweetening capacity due to its content of glycosides is used to reduce lipid profile and this study aimed to decrease lipid profile levels and lowering the risk factor in women using contraceptive drugs by stevia extracts., Materials and Methods: Sixteen rabbits have been used as a case-control study design due to their anatomical and physiological similarity to humans. The stevia leaves are extracted using Soxhlet apparatus of ethanol solvent. Statistical package (SPSS), were used for data analysis and management using independent sample t-test, test, comparison of means for lipid profile of Triglyceride (TG), Cholesterol, High-density lipoprotein (HDL-C), Low-density lipoprotein (LDL-C) between (di-contraceptive, mono-contraceptive and control groups)., Results: The results showed increasing cholesterol and LDL-C during the combined oral contraceptive (COCP) and progesterone-only pills with decreased HDL-C level. A comparison of means before and after stevia used explains the elevated HDL-C and decreased LDL-C., Conclusion: The lipid profile levels should continuously be monitored during oral contraceptive intake and Stevia leaf powder extraction is suggested to reduce the risk of CVD.

Background: Taste and appreciation of sports drinks can affect perceived exertion during exercise. Anecdotal evidence shows that maple products are regularly consumed by recreational and professional athletes but very few studies have reported on their effects during exercise. The purpose of the current study is to report the taste, appreciation and perceived exertion following the ingestion of maple-based sports drinks and other carbohydrate drinks during prolonged exercise., Methods: Recreationally and competitively active male subjects (n = 76, mass = 73.7 ± 10.3 kg, maximum oxygen consumption (VO 2 max) = 4.4 ± 0.5 L/min, maximal aerobic power (MAP) = 309 ± 42 W) ingested one of four carbohydrate solutions (all at 60 g CHO/L): concentrated maple sap (MW), diluted maple syrup (MS), glucose (G), a commercial sports drink (CSD), or a placebo (P; water sweetened with stevia) at every 30 min during 120 min of steady-state exercise (SSE) on a cycle ergometer at 66% MAP. Ratings of perceived exertion (RPE, Borg CR-10) were recorded at each 30 min throughout SSE. A questionnaire was administered to assess sensory characteristics (sweetness, acidity, refreshing, and overall taste on a visual analogue scale, converted to decimals from 0 to 1) and appreciation (sweet, acid and overall on a 9-point hedonic scale) 30 min before (immediately after the first ingestion) and immediately after SSE., Results: Sweetness was perceived to be higher for MW than G and P (pre: 0.60 ± 0.19, 0.51 ± 0.17 and 0.50 ± 0.17 and post: 0.69 ± 0.19, 0.34 ± 0.18 and 0.48 ± 0.22; p < 0.05, respectively) and MS was rated higher than MW for the appreciation of the sweet taste (pre: 6.5 ± 1.5 vs. 4.6 ± 1.8 and post: 6.8 ± 1.8 and 4.1 ± 1.8; p < 0.05, respectively). Furthermore, subjects that had ingested MW, reported a significantly lower RPE than those with P at 120 min (14.1 ± 2.2 vs. 16.0 ± 2.0, respectively)., Conclusions: A sports drink containing maple syrup is well appreciated during prolonged exercise and appears to be a viable alternatives to more common sources of carbohydrates., Trial Registration: NCT02880124 . Registered on 26 August 2016.

Ultrasound-assisted extraction is widely recognized as an eco-friendly technique due to low solvent consumption and time extraction as well as enhanced extraction efficiency with respect to conventional methods. Nevertheless, it would be convenient to avoid the usually used organic solvents to reduce the environment pollution. In this regard, Deep Eutectic Solvents (DES) represent nowadays a green and sustainable alternative for the extraction of bioactive compounds from natural sources. In this study, an efficient extraction of stevioside and rebaudioside A from Stevia rebaudiana coupling ultrasound with DES was developed. A solvent screening was performed using the predictive approach COnductor-like Screening MOdel for Real Solvent (COSMO-RS). The effect of three independent variables, namely % of water, temperature, and sonication amplitude, were investigated by the response surface methodology (RSM). Comparing ultrasound-assisted extraction (UAE) with conventional extraction, it has been demonstrated that the amount of steviol glycosides through UAE is almost three times higher than that obtained by the conventional method. Possible physicochemical factors involved in the UAE mechanism were discussed., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Governments are creating regulations for consumers to reduce their sugar intake, prompting companies to increase the ratio of artificial sweeteners in their products. However, there is evidence of some deleterious effects ascribed to the aforementioned synthetic agents and therefore consumers and food manufacturers have turned their attention to natural dietary sweeteners, such as stevia, to meet their sweetening needs. Stevia is generally considered safe; however, emerging scientific evidence has implicated the agent in gut microbial imbalance. In general, regulation of microbial behavior is known to depend highly on signaling molecules via quorum sensing (QS) pathways. This is also true for the gut microbial community. We, therefore, evaluated the possible role of these stevia-based natural sweeteners on this bacterial communication pathway. The use of a commercial stevia herbal supplement resulted in an inhibitory effect on bacterial communication, with no observable bactericidal effect. Purified stevia extracts, including stevioside, rebaudioside A (Reb A), and steviol revealed a molecular interaction, and possible interruption of Gram-negative bacterial communication, via either the LasR or RhlR receptor. Our in-silico analyses suggest a competitive-type inhibitory role for steviol, while Reb A and stevioside are likely to inhibit LasR-mediated QS in a non-competitive manner. These results suggest the need for further safety studies on the agents.