Germinated Millet (Pennisetum glaucum (L.) R. Br.) Flour Improved the Gut Function and Its Microbiota Composition in Rats Fed with High-Fat High-Fructose Diet.

Germinated millet (Pennisetum glaucum (L.) R. Br.) is a source of phenolic compounds that has potential prebiotic action. This study aims at evaluating the action of germinated pearl millet on gut function and its microbiota composition in Wistar rats fed with a high-fat high-fructose (HFHF) diet. In the first stage, lasting eight weeks, the experiment consisted of two groups: AIN-93M (n = 10) and HFHF group (n = 20). In the second stage, which lasted ten weeks, the animals of the AIN-93M group (n = 10) were kept, while the HFHF group was dismembered into HFHF (HFHF diet, n = 10) and HFHF + millet (HFHF added 28.6% of germinated millet flour, n = 10) groups. After the 18th week, the urine of the animals was collected for the analysis of lactulose and mannitol intestinal permeability by urinary excretion. The histomorphometry was analyzed on the proximal colon and the fecal pH, concentration of short-chain fatty acids (SCFA), and sequencing of microbiota were performed in cecum content. The Mothur v.1.44.3 software was used for data analysis of sequencing. Alpha diversity was estimated by Chao1, Shannon, and Simpson indexes. Beta diversity was assessed by PCoA (Principal Coordinate Analysis). The functional predictive analysis was performed with PICRUSt2 software (version 2.1.2-b). Functional traits attributed to normalized OTU abundance were determined by the Kyoto Encyclopedia of Genes and Genomes (KEGG). In the results, germinated millet flour reduced Oscillibacter genus and Desulfobacterota phylum, while increasing the Eggerthellaceae family. Furthermore, germinated millet flour: increased beta diversity, cecum weight, and cecum/body weight ratio; improved gut histological parameters by increasing the depth and thickness of the crypt and the goblet cell count (p < 0.05); reduced (p < 0.05) the fecal pH and mannitol urinary excretion; increased (p < 0.05) the propionate short-chain fatty acid concentration. Thus, germinated millet has the potential to improve the composition of gut microbiota and the intestinal function of rats fed with an HFHF diet.

Development and validation of a UHPLC-ESI-QTOF mass spectrometry method to analyze opines, plant biomarkers of crown gall or hairy root diseases.

Opines are low-molecular-weight metabolites specifically biosynthesized by agrobacteria-transformed plant cells when plants are struck by crown gall and hairy root diseases, which cause uncontrolled tissue overgrowth. Transferred DNA is sustainably incorporated into the genomes of the transformed plant cells, so that opines constitute a persistent biomarker of plant infection by pathogenic agrobacteria and can be targeted for crown gall/hairy root disease diagnosis. We developed a general, rapid, specific and sensitive analytical method for overall opine detection using ultra-high-performance liquid chromatography-electrospray ionization quadrupole time-of-flight mass spectrometry (UHPLC-ESI-MS-QTOF), with easy preparation of samples. Based on MS, MS/MS and chromatography data, the detection selectivity of a wide range of standard opines was validated in pure solution and in different plant extracts. The method was successfully used to detect different structural types of opines, including opines for which standard compounds are unavailable, in tumors or hairy roots induced by pathogenic strains. As the method can detect a wide range of opines in a single run, it represents a powerful tool for plant gall analysis and crown gall/hairy root disease diagnosis. Using an appropriate dilution of plant extract and a matrix-based calibration curve, the quantification ability of the method was validated for three opines belonging to different families (nopaline, octopine, mannopine), which were accurately quantified in plant tissue extracts.

Comprehensive liamocin biosurfactants analysis by reversed phase liquid chromatography coupled to mass spectrometric and charged-aerosol detection.

Liamocin biosurfactants and structurally related exophilins secreted by the Aureobasidium pullulans (A. pullulans) strain NRRL62031 were firstly analyzed by hyphenation of high-performance liquid chromatography (HPLC) with high-resolution mass spectrometry (HRMS). Ten different analytes were detected and identified by their accurate masses and divided into subclasses according to their different head groups: three liamocins with arabitol as head group, three mannitol liamocins, and four exophilins. A baseline separation of congeners within the subclasses was achieved by reversed phase HPLC on a C18 stationary phase, whereas an overlap of subclasses occurred. The structures were simultaneously confirmed by online tandem mass spectrometry (MS/MS) experiments in positive and negative ionization mode. The assigned polyol head groups and thus the feasibility of this method were confirmed by gas chromatography (GC)-MS data obtained after hydrolysis and derivatization of the liamocins. Based on the varying structural characteristics of liamocins, e.g. the polyol head group (or even none for exophilins) and the degree of acetylation, different detector response in LC-MS was expected, impairing relative quantification of congeners. Therefore, a complementary quantification method was developed using HPLC coupled to charged-aerosol detection (CAD), which allows the determination of the amount of the individual liamocin species without authentic liamocin standards. Hence, the here presented hyphenated techniques facilitate comprehensive analysis of liamocin biosurfactants.

Mannitol: physiological functionalities, determination methods, biotechnological production, and applications.

Mannitol is a naturally occurring six-carbon sugar alcohol that has wide applications in the food and pharmaceutical industry because of its many properties, namely being a natural sweetener with a low metabolism and no glycemic index. The increasing demand for mannitol has spurred many studies of its production. Compared with its chemical synthesis and extraction from plants, both of which are difficult to satisfy for industrial requirements, biotechnological production of mannitol has received considerably more attention and interest from scientists because of its known advantages over those two methods. Accordingly, in this review, we summarize recent advances made in the production of mannitol through various biotechnological methods. The physicochemical properties, sources, and physiological functionalities and applications of mannitol are systematically covered and presented. Then, different determination methods for mannitol are also described and compared. Furthermore, different biotechnological strategies for the production of mannitol via fermentation engineering, protein engineering, and metabolic engineering receive a detailed overview in terms of mannitol-producing strains, enzymes, and their key reaction parameters and conditions. KEY POINTS: • Physiological functionalities and applications of mannitol are presented in detail. • Different determination methods for mannitol are also described and compared. • Various biotechnological strategies for the production of mannitol are reviewed.

1H NMR-based metabolomics for the discrimination of celery (Apium graveolens L. var. dulce) from different geographical origins.

Celery (Apium graveolens L. var dulce) is a widely cultivated vegetable which is popularly consumed due to its nutrient content and contains bioactive metabolites with positive effects on human physiology. In this study, 1H NMR spectroscopy coupled with multivariate statistical analyses was used to distinguish celery stem and leaf samples from different geographical origins. Principal component analysis (PCA) and partial least squares discriminant analysis (PLS-DA) were employed to investigate the differences between celery extracts from three geographical origins: Australia, Taiwan and China. Sugars, amino acids and organic acids were found to contribute significantly to the differentiation between origins, with mannitol identified as an important discriminating metabolite. It was demonstrated that NMR-based metabolomics is an effective approach for establishing reliable metabolomic fingerprints and profiles, enabling the identification of metabolite biomarkers for the possible discrimination of geographical origin.

How Do Trichoderma Genus Fungi Win a Nutritional Competition Battle against Soft Fruit Pathogens? A Report on Niche Overlap Nutritional Potentiates.

We present a case study report into nutritional competition between Trichoderma spp. isolated from wild raspberries and fungal phytopathogenic isolates (Colletotrichum sp., Botrytis sp., Verticillium sp. and Phytophthora sp.), which infect soft fruit ecological plantations. The competition was evaluated on the basis of nutritional potentiates. Namely, these were consumption and growth, calculated on the basis of substrate utilization located on Biolog® Filamentous Fungi (FF) plates. The niche size, total niche overlap and Trichoderma spp. competitiveness indices along with the occurrence of a stressful metabolic situation towards substrates highlighted the unfolding step-by-step approach. Therefore, the Trichoderma spp. and pathogen niche characteristics were provided. As a result, the substrates in the presence of which Trichoderma spp. nutritionally outcompete pathogens were denoted. These were adonitol, D-arabitol, i-erythritol, glycerol, D-mannitol and D-sorbitol. These substrates may serve as additives in biopreparations of Trichoderma spp. dedicated to plantations contaminated by phytopathogens of the genera Colletotrichum sp., Botrytis sp., Verticillium sp. and Phytophthora sp.

Recent advances in the biotechnological production of erythritol and mannitol.

Dietary habits that include an excess of added sugars have been strongly associated with an increased risk of obesity, heart disease, diabetes, and tooth decay. With this association in view, modern food systems aim to replace added sugars with low calorie sweeteners, such as polyols. Polyols are generally not carcinogenic and do not trigger a glycemic response. Furthermore, owing to the absence of the carbonyl group, they are more stable compared to monosaccharides and do not participate in Maillard reactions. As such, since polyols are stable at high temperatures, and they do not brown or caramelize when heated. Therefore, polyols are widely used in the diets of hypocaloric and diabetic patients, as well as other specific cases where controlled caloric intake is required. In recent years, erythritol and mannitol have gained increased importance, especially in the food and pharmaceutical industries. In these areas, research efforts have been made to improve the productivity and yield of the two polyols, relying on biotechnological manufacturing methods. The present review highlights the recent advances in the biotechnological production of erythritol and mannitol and summarizes the benefits of using the two polyols in the food and pharmaceutical industries.

Content of phenolic compounds and mannitol in olive leaves extracts from six Spanish cultivars: Extraction with the Soxhlet method and pressurized liquids.

Olive leaves are considered a promising source of bioactives such as phenolic compounds and mannitol. The extraction of high added value products is an issue of great interest and importance from the point of view of their exploitation. However, the content of these compounds can differ between cultivars and extraction methods. In this work, six olive leaves cultivars, including three wild cultivars, and two extraction processes (an innovative and alternative technique, pressurized liquid extraction, and a conventional Soxhlet extraction) were evaluated and compared towards the selective recovery of bioactive compounds. The wild cultivars showed the highest content of phenolic and flavonoid compounds, being oleuropein the compound present in higher amount. Findings also revealed that the highest mannitol content in the extracts was observed with the commercial cultivars, specifically in Arbequina. It is thus possible to decide which cultivars to use in order to obtain the highest yield of each bioproduct.

Genetic evidences for the core biosynthesis pathway, regulation, transport and secretion of liamocins in yeast-like fungal cells.

So far, it has been still unknown how liamocins are biosynthesized, regulated, transported and secreted. In this study, a highly reducing polyketide synthase (HR-PKS), a mannitol-1-phosphate dehydrogenase (MPDH), a mannitol dehydrogenase (MtDH), an arabitol dehydrogenase (ArDH) and an esterase (Est1) were found to be closely related to core biosynthesis of extracellular liamocins in Aureobasidium melanogenum 6-1-2. The HR-PKS was responsible for biosynthesis of 3,5-dihydroxydecanoic acid. The MPDH and MtDH were implicated in mannitol biosynthesis and the ArDH was involved in arabitol biosynthesis. The Est1 catalyzed ester bond formation of them. A phosphopantetheine transferase (PPTase) activated the HR-PKS and a transcriptional activator Ga11 activated expression of the PKS1 gene. Therefore, deletion of the PKS1 gene, all the three genes encoding MPDH, MtDH and ArDH, the EST1, the gene responsible for PPTase and the gene for Ga11 made all the disruptants (Δpks13, Δpta13, Δest1, Δp12 and Δg11) totally lose the ability to produce any liamocins. A GLTP gene encoding a glycolipid transporter and a MDR1 gene encoding an ABC transporter took part in transport and secretion of the produced liamocins into medium. Removal of the GLTP gene and the MDR1 gene resulted in a Δgltp1 mutant and a Δmdr16 mutant, respectively, that lost the partial ability to secrete liamocins, but which cells were swollen and intracellular lipid accumulation was greatly enhanced. Hydrolysis of liamocins released 3,5-dihydroxydecanoic acid, mannitol, arabitol and acetic acid. We proposed a core biosynthesis pathway, regulation, transport and secretion of liamocins in A. melanogenum.

Sourdough technology as a novel approach to overcome quality losses in sugar-reduced cakes.

Sugar reduction in sweet baked goods is one of the most popular trends on the food market. However, reducing sugar without lowering the product quality with respect to sweetness, texture and microbial shelf life is challenging. Sugar alcohols are one group of sugar replacers which maintain the bulk and contribute to sweetness. Nevertheless, alternative approaches, particularly those that are seen as 'clean label', are highly demanded. Hence, the natural, in situ production of mannitol in a sourdough system was performed and its potential as a functional ingredient to improve the product quality of a sugar-reduced cake was investigated. A full sugar cake (C1), a 50% sugar-reduced cake with wheat starch (C2) or with commercially available mannitol (C3) were considered as controls. The substitution of sugar by wheat starch or mannitol caused lower specific volume (-15.5%; -10.7%), a harder crumb (+17.1 N; +4.4 N), less browning, a shorter microbial shelf life (-5.7 days; -4.3 days) and poorer sensory properties. The incorporation of sourdough in C2 improved the pasting properties by decreasing the peak viscosity. Moreover, it resulted in a similar specific volume to the full-sugar control, contributed to significantly softer crumb (-8.6 N) and increased browning compared to C2. Sensory evaluation revealed an increased sweetness perception (+93%), aroma (+30%) and flavour (+25.5%) by the incorporation of sourdough. Although sourdough incorporation improved sugar-reduced cakes, the quality characteristics of the full-sugar control could not be achieved in this study. In conclusion, the addition of sourdough in amounts lower than 10% can be considered a useful tool to improve specific volume, crumb structure, colour, taste and flavour, as well as shelf life of sugar-reduced cakes.

1H-NMR-Based Metabolic Profiling of Cordyceps militaris to Correlate the Development Process and Anti-Cancer Effect.

The study of metabolomics in natural products using the diverse analytical instruments including GC-MS, LC-MS, and NMR is useful for the exploration of physiological and biological effects and the investigation of drug discovery and health functional foods. Cordyceps militaris has been very attractive to natural medicine as a traditional Chinese medicine, due to its various bioactive properties including anti-cancer and anti-oxidant effects. In this study, we analyzed the metabolite profile in 50% ethanol extracts of C. militaris fruit bodies from three development periods (growth period, matured period, and aging period) using 1H-NMR, and identified 44 metabolites, which are classified as 16 amino acids, 10 organic acids, 5 carbohydrates, 3 nucleotide derivatives, and 10 other compounds. Among the three development periods of the C. militaris fruit body, the aging period showed significantly higher levels of metabolites including cordycepin, mannitol (cordycepic acid), and ß-glucan. Interestingly, these bioactive metabolites are positively correlated with antitumor growth effect; the extract of the aging period showed significant inhibition of HepG2 hepatic cancer cell proliferation. These results showed that the aging period during the development of C. militaris fruit bodies was more highly enriched with bioactive metabolites that are associated with cancer cell growth inhibition.

Analysis of Genetic Association of Intestinal Permeability in Healthy First-degree Relatives of Patients with Crohn's Disease.

Excessive intestinal permeability or intestinal barrier dysfunction as measured by various assays has been observed in various diseases. However, little is known about the factors contributing to altered gut permeability in these diseases. Our objective was to determine the genetic determinants of altered gut permeability as measured by the lactulose mannitol fractional excretion ratio (LacMan ratio) in 1075 healthy first-degree relatives of patients with Crohn's disease (CD). In a targeted analysis of single nucleotide polymorphisms (SNPs) located in genes associated with intestinal barrier function related or not to inflammatory bowel disease, we did not find a significant association with intestinal permeability. In an untargeted genome-wide association analysis, the top 100 associations were located in 22 genomic loci, although they were not statistically significant after correction for multiple testing (raw P values [1.8 × 10-7 - 1.4 × 10-5]. The lowest P value was obtained for rs9616637 (22q13.33, C22orf34), for which the minor allele A was associated with a decreased LacMan ratio. These results suggest that host genetic background has limited contribution toward intestinal permeability. Despite this, our study is currently the largest of its kind assessing gut permeability in vivo. It remains possible that smaller genetic effect sizes on LacMan ratio are not detectable in this sized cohort. Larger studies are warranted to identify the potential genetic contribution to intestinal permeability.

Combinatorial impact of osmotic and heat shocks on the composition of membrane lipids and osmolytes in Aspergillus niger.

The combinatorial action of osmotic (OS) and heat (HS) shocks on the composition of soluble cytosol carbohydrates and membrane lipids was studied. For the first time it was demonstrated that the combinatorial effect of these shocks led to the non-additive response - an increase in the trehalose level, characteristic for HS, but at the same time suppression of glycerol production, uncharacteristic of the OS response. In addition, combinatorial action resulted in a new effect - increase in the mannitol level, which was not typical for the individual HS or OS responses. On the contrary, a general pattern of change was observed in the composition of membrane lipids in response to both individual HS and OS, and their combinations, which was a twofold increase in the proportion of phosphatidic acids. At the same time, the mechanism of alteration in the degree of unsaturation of membrane phospholipids was not involved in adaptation. The response to combinatorial shocks includes the accumulation of trehalose and mannitol, and increase in the proportion of phosphatidic acids in membrane lipids.

Zinc Absorption and Endogenous Fecal Zinc Losses in Bangladeshi Toddlers at Risk for Environmental Enteric Dysfunction.

OBJECTIVES: Environmental enteric dysfunction (EED) impairs zinc absorption from food, and zinc deficiency may contribute to the poor growth associated with EED. We examined zinc absorption from a standardized aqueous zinc dose, and habitual daily endogenous fecal zinc excretion (EFZ) and compared these outcomes between children grouped by the lactulose to mannitol ratio (L:M). METHODS: Bangladeshi toddlers (18-24 months) with low (<0.09) and high (≥0.09) L:M were administered isotope-labeled 3 mg aqueous zinc in the fasted state. Fractional absorption of zinc (FAZ) and EFZ were measured by dual stable isotope tracer method and an isotope dilution method, respectively. Secondary aims included examining relationships of biomarkers of systemic and intestinal inflammation and gut function with FAZ and EFZ. RESULTS: Forty children completed the study; nearly all had evidence of EED. No differences in zinc homeostasis measurements (mean ±â€ŠSD) were observed between high and low L:M groups: FAZ was 0.38 ±â€Š0.19 and 0.31 ±â€Š0.19, respectively; both figures were within estimated reference range. Means of EFZ were 0.73 ±â€Š0.27 and 0.76 ±â€Š0.20 mg/day for high and low L:M, respectively, and were 10% to 15% above estimated reference range. Regression analyses indicated that biomarkers of systemic inflammation were directly associated with increasing FAZ, consistent with increased gut permeability. Biomarkers of intestinal inflammation were negatively associated with EFZ, consistent with low-zinc intake and chronic deficiency. CONCLUSIONS: In these children at risk of EED, endogenous zinc losses were not markedly increased. Results suggest that efforts to improve zinc status in EED should focus on substantially improving zinc intakes.

Valorization of Crude Glycerol, Residue Deriving from Biodiesel- Production Process, with the Use of Wild-type New Isolated Yarrowia lipolytica Strains: Production of Metabolites with Pharmaceutical and Biotechnological Interest.

BACKGROUND & OBJECTIVE: Crude glycerol (Glol), used as substrate for screening eleven natural Yarrowia lipolytica strains in shake-flask experiments. Aim of this study was to assess the ability of the screened strains to produce biomass (dry cell weight; X), lipid (L), citric acid (Cit), mannitol (Man), arabitol (Ara) and erythritol (Ery), compounds presenting pharmaceutical and biotechnological interest, in glycerol-based nitrogen-limited media, in which initial glycerol concentration had been adjusted to 40 g/L. METHODS: Citric acid may find use in biomedical engineering (i.e. drug delivery, tissue engineering, bioimaging, orthopedics, medical device coating, wound dressings). Polyols are considered as compounds with non-cariogenic and less calorigenic properties as also with low insulin-mediated response. Microbial lipids containing polyunsaturated fatty acids (PUFA) are medically and dietetically important (selective pharmaceutical and anticancer properties, aid fetal brain development, the sight function of the eye, hormonal balance and the cardio-vascular system, prevent reasons leading to type-2 diabetes, present healing and anti-inflammatory effects). RESULTS: All strains presented satisfactory microbial growth (Xmax=5.34-6.26 g/L) and almost complete substrate uptake. The principal metabolic product was citric acid (Citmax=8.5-31.7 g/L). Production of cellular lipid reached the values of 0.33-0.84 g/L. Polyols were also synthesized as strain dependent compounds (Manmax=2.8-6.1 g/L, Aramax ~2.0 g/L, Erymax= 0.5-3.8 g/L). The selected Y. lipolytica strain ACA-DC 5029 presented satisfactory growth along with synthesis of citric acid and polyols, thus, was further grown on media presenting an increased concentration of Glol~75 g/L. Biomass, lipid and citric acid production presented significant enhancement (Xmax=11.80 g/L, Lmax=1.26 g/L, Citmax=30.8 g/L), but conversion yield of citric acid produced per glycerol consumed was decreased compared to screening trials. Erythritol secretion (Erymax=15.6 g/L) was highly favored, suggesting a shift of yeast metabolism from citric acid accumulation towards erythritol production. Maximum endopolysaccharides (IPS) concentration was 4.04 g/L with yield in dry weight 34.2 % w/w. CONCLUSION: Y. lipolytica strain ACA-YC 5029 can be considered as a satisfactory candidate grown in high concentrations of crude glycerol to produce added-value compounds that interest pharmaceutical and biotechnology industries.

Effect of the solar drying process on the sensory and chemical quality of cocoa (Theobroma cacao L.) cultivated in Antioquia, Colombia.

The quality of Theobroma cacao L. is influenced by different variables both in the crop and in the processes of postharvest, the latter it includes the fermentation and the drying, fundamental for the formation of aroma and flavor precursors, determinants in the characteristics of quality and differentiation of cocoa in the chocolate industry. The objective of the present research was to evaluate the effect of the solar drying process using a plastic roof solar dryer on the sensory and chemical quality of a mixture of cocoa cultivated in a region of Antioquia, Colombia. The content of total polyphenols, anthocyanins, sugars, mannitol, pH, total acidity, humidity, ethereal extract, ash, crude fiber and odor and flavor descriptors were analyzed. For analyzing the data, we used the methodology of longitudinal data analysis and repeated measurements, a Principal Component Analysis (PCA), and a principal factor analysis. The analyzed sensory characteristics are statistically different over time (p < 0.05). There was a decrease in the content of anthocyanins, total polyphenols and sucrose, and an increase in glucose and fructose during fermentation and drying. The analysis of factors allowed to define a series of groupings as indexes of quality according to the chemical and sensory properties analyzed in the drying process. Overall, the mixture of cocoa clones evaluated in the solar drying process presented indicators of sensory and chemical quality associated with descriptions of odor (spicy, dairy, nut, fruity, sweet cane), flavor (floral and spicy), ethereal extract, and pH that indicate a good benefit of cocoa and show the potential that Colombia has as a producer of fine aroma cocoa in high demand in premium markets.

Metabolomic profiling reveals enrichment of cordycepin in senescence process of Cordyceps militaris fruit bodies.

Cordyceps militaris is a species of Cordyceps that is classified in the Cordycipitaceae family and is well known in East Asia as a traditional medicinal mushroom. Its artificial fruit body has been widely cultivated for commercial use in cosmetics, functional food, and medicine. To explore the metabolites associated with fruit body development, we conducted gas chromatography mass spectrometry (GC-MS) analyses based on developmental stage, which was divided into the growth period (stage 1, stage 2, and stage 3) and aging period (stage 4). We detected 39 biochemical metabolites associated with nucleotide, carbohydrate, and amino acid metabolism. Cordycepin, one of the representative bioactive compounds in C. militaris, was significantly enriched in stage 4 of aging period and is associated with glucose accumulation. The accumulation of cordycepin in stage 4 of aging period also seems to be related to the glutamine and glutamic acid pathway. Our results also showed enrichment of other bioactive compounds such as mannitol and xylitol in stage 4 of aging period. Our metabolomic profiling based on the developmental stages of C. militaris is useful for exploring bioactive compounds (e.g., cordycepin, mannitol, GABA, and xylitol) that are enriched in stage 4 of aging period and understanding the biosynthetic mechanisms associated with cordycepin production. Through optimization of fruit body cultivation by selecting stage 4 of aging period as a harvesting time, our findings can be utilized in food and medical applications of C. militaris in future.

Dynamic Analysis of Nucleosides and Carbohydrates during Developmental Stages of Cordyceps militaris in Silkworm (Bombyxmori).

Background: Cultured Cordyceps militaris is very popular. Objective: To gain dynamic insight into activity markers in fruiting body of Cordyceps militaris (C. militaris) in Bombyxmori (B. mori), also named silkworm. Methods: The development stages of samples at 3, 9, 12, 19, 27, and 33 days after inoculation (DAI) were collected. HPLC coupled with diode array detection and evaporative light-scattering detection method (HPLC-DAD-ELSD) was used to determine eight makers, including six nucleosides and two carbohydrates from the samples. Results: C. militaris cultured 33 DAI with fifth star silkworm larva could accumulate higher levels of cordycepin (13.43 mg/g) than the highest reported cordycepin (8.57 g/L). The contents of cordycepin, adenosine, and trehalose were gradually increased with the formation of C. militaris fruiting bodies on silkworm larva, while mannitol was decreased. The change of guanosine was similar to uracil. Conclusions: Results suggested that mannitol could be accumulated in a short period during mycelium growth and could metabolize and transform into energy store and trehalose during fruit body formation. The inosine in the insect was completely utilized and transformed. The synergistic formation of cordycepin and adenosine or differences in metabolized pathways are a great possibility according to the same trend. Highlights: This research offered some reference to further find a certain regularity or metabolic mechanism.

Qualitative and quantitative analysis of 2, 5-anhydro-d-mannitol in low molecular weight heparins with high performance anion exchange chromatography hyphenated quadrupole time of flight mass spectrometry.

Low molecular weight heparins (LMWHs), derived from unfractionated heparin (UFH) by chemical or enzymatic degradation, have lower side effects than that of heparin. Enoxaparin, nadroparin, and dalteparin are the most widely used LMWHs. Nadroparin and dalteparin are prepared through nitrous acid degradation followed by a subsequent reduction process, in which specific residue, 2,5-anhydro-d-mannitol (An-Man), is formed at the reducing terminals of generated sugar chains. However, few practicable analytical method was available to analyze An-Man qualitatively and quantitatively. In this work, a HPAEC-PAD-MS method was developed to analyze monosaccharides in heparin and LMWHs, especially An-Man. An ion suppressor is set up between HPAEC and MS to remove the nonvolatile salts from HPAEC and make the elute compatible to MS. Various monosaccharides were separated well with HPAEC. Online MS and MS/MS confirmed all sugar residues in the hydrolysates of heparin samples. The specific residue, An-Man, was only observed in the hydrolysates of LMWHs prepared with nitrous acid degradation and reduction. In addition, major glucosamine and minor arabinose/galactose were observed in all heparin samples. The amounts of these sugar residues were quantitated with PAD, simultaneously. The ratio of glucosamine to An-Man could be used to calculate the molecular weight of LMWHs. The calculated values are comparable to the value measured with size-exclusion chromatography-multiple angle laser scattering detection. Higher the ratio of glucosamine to An-Man higher the molecular weight. The HPAEC-PAD-MS platform is an accurate, precise and efficient way to identify LMWHs by determination of An-Man. It is also an alternative method to detect the MWs of LMWHs having An-Man for quality control purposes.

Xylitol, mannitol and maltitol as potential sucrose replacers in burger buns.

Burger buns are a source of added sugar, containing 7-12%, in order to ensure their unique texture and taste. Hence, suitable sugar substitutes for burger buns are urgently needed. This study aimed to elucidate the effect of three different polyols on dough and product quality of burger buns. Xylitol, mannitol and maltitol were incorporated individually in a burger bun system, by replacing added sucrose by 30%, 50% and 100%. Wheat starch was used to compare the impact of polyols with another non-sweet bulking agent. The effects on dough properties as well as on the burger buns themselves were investigated. Compared to sugar-rich doughs, polyols lowered the fermentation quality, resulting in lower dough development (-37 to -81%) and poorer gaseous release (-62 to -87%). Furthermore, a delay in gluten network development (+50 to +161%) and a decrease in extensibility (-14 to -18%) with increasing concentrations were detected. Interestingly, maltitol and xylitol did not affect the pasting properties, whereas mannitol increased pasting temperature (+15 °C). Moreover, polyols did not influence the viscoelastic properties of the dough. The incorporation of sugar alcohols led to a significant decrease in specific volume (-30 to -48%), and to a harder crumb texture (+135 to +678%). Moreover, the L*-value increased with increasing amount of polyols, resulting in a very pale crust colour. In conclusion, a reduction of 50% added sucrose by polyols was applicable, whereas mannitol was the most suitable sugar replacer amongst the polyols tested.