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Background Sugar alcohols are widely used as low-calorie sweeteners in the food and pharmaceutical industries. They can also be transformed into platform chemicals. Yarrowia lipolytica, an oleaginous yeast, is a promising host for producing many sugar alcohols. In this work, we tested whether heterologous expression of a recently identified sugar alcohol phosphatase (PYP) from Saccharomyces cerevisiae would increase sugar alcohol production in Y. lipolytica. Results Y. lipolytica was found natively to produce erythritol, mannitol, and arabitol during growth on glucose, fructose, mannose, and glycerol. Osmotic stress is known to increase sugar alcohol production, and was found to significantly increase erythritol production during growth on glycerol. To better understand erythritol production from glycerol, since it was the most promising sugar alcohol, we measured the expression of key genes and intracellular metabolites. Osmotic stress increased the expression of several key genes in the glycerol catabolic pathway and the pentose phosphate pathway. Analysis of intracellular metabolites revealed that amino acids, sugar alcohols, and polyamines are produced at higher levels in response to osmotic stress. Heterologous overexpression of the sugar alcohol phosphatase increased erythritol production and glycerol utilization in Y. lipolytica. We further increased erythritol production by increasing the expression of native glycerol kinase (GK), and transketolase (TKL). This strain was able to produce 27.5 ± 0.7 g/L erythritol from glycerol during batch growth and 58.8 ± 1.68 g/L erythritol during fed-batch growth in shake-flasks experiments. In addition, the glycerol utilization was increased by 2.5-fold. We were also able to demonstrate that this strain efficiently produces erythritol from crude glycerol, a major byproduct of the biodiesel production. Conclusions We demonstrated the application of a promising enzyme for increasing erythritol production in Y. lipolytica. We were further able to boost production by combining the expression of this enzyme with other approaches known to increase erythritol production in Y. lipolytica. This suggest that this new enzyme provides an orthogonal route for boosting production and can be stacked with existing designs known to increase sugar alcohol production in yeast such as Y. lipolytica. Collectively, this work establishes a new route for increasing sugar alcohol production and further develops Y. lipolytica as a promising host for erythritol production from cheap substrates such as glycerol. more...

Aims To assess the ability of various newly isolated or belonging in official collections yeast strains to convert biodiesel-derived glycerol (Gly) into added-value compounds. Methods and results Ten newly isolated yeast strains belonging to Debaryomyces sp., Naganishia uzbekistanensis, Rhodotorula sp. and Yarrowia lipolytica, isolated from fishes, metabolized Gly under nitrogen limitation. The aim of the study was to identify potential newly isolated microbial candidates that could produce single-cell oil (SCO), endopolysaccharides and polyols when these micro-organisms were grown on biodiesel-derived Gly. As controls producing SCO and endopolysaccharides were the strains Rhodotorula glutinis NRRL YB-252 and Cryptococcus curvatus NRRL Y-1511. At initial Gly (Gly0 ) ≈40 g l-1 , most strains presented remarkable dry cell weight (DCW) production, whereas Y. lipolytica and Debaryomyces sp. produced non-negligible quantities of mannitol and arabitol (Ara). Five strains were further cultivated at increasing Gly0 concentrations. Rhodotorula glutinis NRRL YB-252 produced 7·2 g l-1 of lipid (lipid in DCW value ≈38% w/w), whereas Debaryomyces sp. FMCC Y69 in batch-bioreactor experiment with Gly0 ≈80 g l-1 , produced 30-33 g l-1 of DCW and ~30 g l-1 of Ara. At shake-flasks with Gly0 ≈125 g l-1 , Ara of ~48 g l-1 (conversion yield of polyol on Gly consumed ≈0·62 g g-1 ) was achieved. Cellular lipids of all yeasts contained in variable concentrations oleic, palmitic, stearic and linoleic acids. Conclusions Newly isolated, food-derived and non-previously studied yeast isolates converted biodiesel-derived Gly into several added-value metabolites. Significance and impact of the study Alternative ways of crude Gly valorization through yeast fermentations were provided and added-value compounds were synthesized. more...

Polyol dehydrogenases (PDHs) play a pivotal role in the biotransformation between rare sugar and alcohol. Among these PDHs, mannitol 2-dehydrogenase (MDH, EC 1.1.1.67), galactitol 2-dehydrogenase (GDH, EC 1.1.1.16), ribitol 2-dehydrogenase (RDH, EC 1.1.1.56), xylitol 4-dehydrogenase (XDH, EC 1.1.1.14), and arabitol 2-dehydrogenase (ArDH, EC 1.1.1.12) are the most studied. MDH can catalyze the transformation between D-fructose and mannitol as well as the transformation between D-arabitol and D-xylulose. In addition to MDH, the other PDHs including RDH, GDH, ArDH, and XDH are also important tools for the production of rare sugars including D-tagatose, allitol, D-xylulose, and L-xylulose. Concerning the intermediate function of PDH in the linkage of rare sugar and sugar alcohols, this review attempts to conclude their catalytic properties and potential applications. more...

Size-segregated aerosol samples collected from a rural site (Rajim: 20˚59′N and 81˚55′E) in eastern central India during the winter of 2011 were analyzed for anhydrosugars, sugar alcohols and primary sugars to better understand their potential sources and formation. Anhydrosugars showed the predominance of levoglucosan, whereas sugar alcohol showed the predominance of mannitol, with glucose as a major primary sugar in the sub-micron (Da 1.1 μm). Levoglucosan gave a bimodal size distribution pattern with a major peak at 0.43–0.65 μm in the sub-micron mode during period 2 and period 3, suggesting its origin from biomass burning, whereas during period 1 the major peak was found at 4.4–5.8 μm in super-micron mode that may be due to the mixing of biomass burning derived particles with soil dust particles. Sugar alcohols (arabitol and mannitol) and primary sugars (glucose and trehalose) showed bimodal size distribution trend with a major peak at 4.4–5.8 μm in super-micron mode during all periods, perhaps due to fungal activity and soil dust re-suspension, respectively. The potential ingress of biomass burning derived particles into the lungs was calculated to be high whereas fungal spores and soil particles were more likely found in the upper respiratory tract. Principal component analysis and correlation analysis showed that biomass burning was a major source in the sub-micron mode whereas fungal spores and vegetative soil were dominant sources for quantified sugars in super-micron mode. more...

Liamocins are biosurfactants produced by the fungus Aureobasidium pullulans. A. pullulans belongs to the black yeasts and is known for its ability to produce pullulan and melanin. However, the production of liamocins has not been investigated intensively. Initially, HPLC methods for the quantification of liamocin and the identification of liamocin congeners were established. Eleven congeners could be detected, differing in the polyol head groups arabitol or mannitol. In addition, headless molecules, so-called exophilins, were also identified. The HPLC method reported here allows quick and reliable quantification of all identified congeners, an often-overlooked prerequisite for the investigation of valuable product formation. Liamocin synthesis was optimized during cultivation in lab-scale fermenters. While the pH can be kept constant, the best strategy for liamocin synthesis consists of a growth phase at neutral pH and a subsequent production phase induced by a manual pH shift to pH 3.5. Finally, combining increased nitrogen availability with a pulsed fed-batch fermentation, cell growth, and liamocin titers could be enhanced. Here, the maximal titers of above 10 g/L that were reached are the highest reported to date for liamocin synthesis using A. pullulans in lab-scale fermenters. more...

A new yeast wild-type Yarrowia lipolytica isolate presented efficient growth on glycerol. During flask cultures, nitrogen limitation led to the secretion of sugar-alcohols as the major metabolites of the process (mannitol, arabitol and erythritol), whereas insignificant quantities of citrate were synthesized. Although in some instances high initial glycerol concentrations were employed (&asymp, 150 g/L), remarkable glycerol assimilation and polyol secretion was observed. Total polyols &asymp, 52 g/L (conversion yield on glycerol consumed = 0.43 g/g) was recorded in the flask experiments. The sugar-alcohol production bioprocess was successfully simulated with the aid of a modified Velhlust&ndash, Aggelis model that fitted very well with the experimental data, while optimized parameter values seemed to be quite consistent. In bioreactor trials, a noticeable metabolic shift towards citric acid production was observed, while simultaneously insignificant polyol quantities were produced. In fed-batch bioreactor experiments, a total citric acid quantity &asymp, 102 g/L was recorded&mdash, one of the highest in the literature for wild-type Y. lipolytica strains. This metabolic transition was due to higher oxygen saturation into the medium that occurred in the bioreactor experiments compared with the flasks. Cellular lipids produced in the bioreactor trial contained higher concentrations of unsaturated fatty acids compared with those produced in flasks. more...

Arabitol has several applications in food and pharmaceutical industries as a natural sweetener, dental caries inhibitor, and texturing agent. Newly isolated yeast strains from seawater, sugarcane plantation soil samples, and Zygosaccharomyces rouxii 2635 from MTCC were tested for arabitol production. The yield of arabitol was found to be higher in seawater isolate (24.6 g L-1 ) compared to two soil isolates (22.5 g L-1 ) and Z. rouxii (19.4 g L-1 ). Based on ITS 26S rDNA sequence analysis, the seawater isolate was identified as Pichia manchurica. In the present study, the effect of different substrates, trace elements, nitrogen sources, pH, and temperature on arabitol production was examined. Three different carbon sources viz. glucose, arabinose, and galactose were studied. Glucose was determined to be the best substrate for arabitol production (27.6 g L-1 ) followed by arabinose (13.7 g L-1 ) and galactose (7.7 g L-1 ). Maximum production of arabitol was observed at pH 6.0 (34.7 g L-1 ). In addition, arabitol production was high (35.7 g L-1 ) at temperature of 30 °C. Among the different concentrations of ammonium sulfate tested (3, 4.5, 6, 7.5, and 9 g L-1 ) concentration of 6 g L-1 resulted in higher arabitol Individual metal ions had no effect on arabitol production by this strain as compared to control. Results obtained in this study identify ways for improved arabitol production with natural isolates using microbial processes. more...

In this study, we developed a simple and sensitive reversed phase ion-pair chromatographic method for the analysis of C4-C6 sugar alcohols. The method is based on the on-line complexation of sugar alcohols with molybdate ion. The resulting dinuclear anionic complexes can be separated on a reversed-phase C18 column using tetrabutylammonium chloride as an ion-pairing reagent. The mobile phase (pH 3.1) consisted of 0.1 mM disodium molybdate, 1 mM hydrochloric acid and 0.4 mM tetrabutylammonium chloride – 10% v/v methanol. By complexing with molybdate ion, sugar alcohols can be detected by their UV absorption at 247 nm with high resolution and sensitivity. The quantification limits of the examined sugar alcohols calculated at S/N = 10 were 0.1 mM for erythritol and xylitol and 0.01 mM for arabitol, sorbitol, mannitol and dulcitol. The detector response was linear over three orders of magnitude of sugar alcohol concentration. The proposed method was successfully applied to measure sugar alcohols in health drinks, eyedrops and mouthwashes. more...

Sugar alcohols are widely marketed compounds. They are useful building block chemicals and of particular value as low- or non-calorigenic sweeteners, serving as sugar substitutes in the food industry. To date most sugar alcohols are produced by chemical routes using pure sugars, but a transition towards the use of renewable, non-edible feedstocks is anticipated. Several yeasts are naturally able to convert renewable feedstocks, such as lignocellulosic substrates, glycerol and molasses, into sugar alcohols. These bioconversions often face difficulties to obtain sufficiently high yields and productivities necessary for industrialization. This review provides insight into the most recent studies on utilizing yeasts for the conversion of renewable feedstocks to diverse sugar alcohols, including xylitol, erythritol, mannitol and arabitol. Moreover, metabolic approaches are highlighted that specifically target shortcomings of sugar alcohol production by yeasts from these renewable substrates. more...

Arabitol is a low-calorie sugar alcohol with anti-cariogenic properties. Enzymatic hydrolysate of soybean flour is a new renewable biorefinery feedstock containing hexose, pentose, and organic nitrogen sources. Arabitol production by Debaryomyces hansenii using soybean flour hydrolysate was investigated. Effects of medium composition, operating conditions, and culture stage (growing or stationary phase) were studied. Production was also compared at different culture volumes to understand the effect of dissolved oxygen concentration (DO). Main factors examined for medium composition effects were the carbon to nitrogen concentration ratio (C/N), inorganic (ammonium) to organic nitrogen ratio (I/O-N), and sugar composition. Arabitol yield increased with increasing C/N ratio and a high I/O-N (0.8–1.0), suggesting higher yield at stationary phase of low pH (3.5–4.5). Catabolite repression was observed, with the following order of consumption: glucose > fructose > galactose > xylose > arabinose. Arabitol production also favored hexoses and, among hexoses, glucose. DO condition was of critical importance to arabitol production and cell metabolism. The yeast consumed pentoses (xylose and arabinose) only at more favorable DO conditions. Finally, arabitol was produced in fermentors using mixed hydrolysates of soy flour and hulls. The process gave an arabitol yield of 54%, volumetric productivity of 0.90 g/L-h, and specific productivity of 0.031 g/g-h. more...

Quantification of carbohydrates and metabolites in fermentation processes can be considered as key factor in determining yield and productivity for a better understanding of the microbial behavior under different conditions. The main aim of the present study was to develop and set up analytical methods for detecting complex sugar and/or metabolite mixtures in fermentation broth based on high-performance thin-layer chromatography (HPTLC). HPTLC is a fast and accurate method of separating complex mixtures, based on planar development. The proposed methods involved the separations of a mixture of monosaccharides (glucose, xylose, arabinose, and rhamnose) deriving from delignification and hydrolysis of hazelnut shells and the corresponding sugar alcohols (xylitol, arabitol, and sorbitol) obtained by fermentations of Candida tropicalis spp., a yeast able to ferment aldoses to produce sugar alcohols. HPTLC methods were set up on simple chamber development and using instrumental techniques like overpressured lay... more...

Liamocins are a class of biosurfactants with growing interest. However, methods for identification and quantification of liamocins on the molecular level are lagging behind. Therefore, we developed a chromatographic separation based on supercritical fluid chromatography (SFC) for liamocins and structurally related exophilins. The different congeners could be separated on a charge modulated hydroxyethyl amide functionalized silica-based column. Coupling to high-resolution mass spectrometry (MS) revealed four exophilin species and four liamocin species with mannitol and arabitol as polyol head group in a sample of the yeast-like fungus Aureobasidium pullulans (A. pullulans). In contrast to a recently published reversed phase high-performance liquid chromatography (HPLC) method, the different subclasses (exophilins, mannitol liamocins and arabitol liamocins) were additionally separated by means of SFC. The structures were confirmed by their accurate masses and tandem mass spectrometry (MS/MS). A complementary quantification method was developed using SFC coupled to charged-aerosol detection (CAD) to overcome the disadvantages of quantification by means of MS without authentic standards. A flow compensation by varying the make-up flow was used to obtain a constant composition of the mobile phase during detection and to ensure a stable detector response. The concentrations of the individual liamocin species were determined using an external calibration with n-octyl-β-d-glycopyranoside. The total amount of these concentrations agrees with the dry weight of an aliquot of the heavy oil. The developed SFC-based method has the advantage of shorter analysis time and superior selectivity compared to the previously published LC separation. In brief, the here presented SFC hyphenations enable comprehensive analysis of liamocin biosurfactants providing identification and absolute quantification of individual congeners. more...

Metabolites of serum and milk from genetically modified (GM) cows and contrast check (CK) cows were comparatively investigated. Serum and milk were collected from genetically modified (GM) cows and contrast check (CK) cows, and then, they were analyzed using ultraperformance liquid chromatography-mass spectrometry (UPLC-MS) and gas chromatography-mass spectrometry (GC-MS). Although the level of some blood biochemical indexes for GM cows was shifted up or down, they were generally in normal physiological condition. Serum samples from lactoferrin GM cows exhibited reduced levels of amino acids and elevated levels of indoleacetate, α-keto acids, long-chain fatty acids, etc. GM milk possessed elevated levels of pentose and amino sugar metabolites, including arabitol, xylulose, glucuronate, and N-acetylgalactosamine. Interestingly, some essential nutrients, such as certain unsaturated fatty acids (e.g., eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA), and docosapentaenoic acid (DPA)), and some necessary rare sugars were significantly upregulated. Compared to the CK group, a Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis was conducted based on the increased or decreased metabolites identified in the serum and milk samples of the GM group. The results showed that the GM cows were in healthy condition and their milk has improved benefits for customers. The milk from genetically modified cows was found to be a promising milk source for producing recombinant human lactoferrin (rhLF) for human beings. more...

Made available in DSpace on 2019-10-06T16:25:30Z (GMT). No. of bitstreams: 0 Previous issue date: 2019-06-01 Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) The present work describes an easy, rapid and sensitive alternative methodology for simultaneous detection of sugars (glucose, fructose and arabinose), organic acids (gluconic, glucuronic and glucaric acids) and sugar alcohol (arabitol) in wine samples. The method is based on high performance anion-exchange chromatography with pulsed amperometric detection (HPAEC-PAD) at gold electrode under detection potential of 0.26 V vs Pd|PdO. The optimized chromatographic anion-exchange separation indicates good separation and detection using a binary gradient with 0.28 mol L −1 CH 3 COONa containing 0.10 mol L −1 NaOH and ultrapure water as mobile phase with a flow rate at 1.0 mL min −1 . Analytical curves presented linear relationship from 1.0 × 10 −7 and 1.0 × 10 −3 mol L −1 with detection limits (S/N = 3) from 3.0 × 10 −8 (glucose) to 9.0 × 10 −7 mol L −1 (gluconic acid). The present method was successfully applied in the determination of sugars, organic acids and sugar alcohols in wine, without time consuming pre-treatment of the sample, no need for organic solvent, fast relative execution time, satisfactory separation, high sensitivity and accuracy. São Paulo State University (Unesp) Institute of Chemistry Analytical Chemistry Department National Institute of Alternative Technologies for Detection Toxicological Assessment and Removal of Micropollutants and Radioactive (INCT-DATREM), Professor Francisco Degni Street 55 São Paulo State University (Unesp) Bioenergy Research Institute, Professor Francisco Degni Street 55 School of Industrial Engineering Metallurgical Volta Redonda Fluminense Federal University (UFF), Trabalhadores Avenue 420 São Paulo State University (Unesp) Institute of Chemistry Analytical Chemistry Department National Institute of Alternative Technologies for Detection Toxicological Assessment and Removal of Micropollutants and Radioactive (INCT-DATREM), Professor Francisco Degni Street 55 São Paulo State University (Unesp) Bioenergy Research Institute, Professor Francisco Degni Street 55 CNPq: 170176/2017-7 FAPESP: 2014/03679-7 FAPESP: 2015/12998-1 CNPq: 465571/2014-0 more...

Summary In this research, the effects of culture substrates on taste component content of Lentinula edodes were studied, and the resulting taste quality of L. edodes was evaluated. The results revealed that single-carbon and single-nitrogen sources were beneficial to production of soluble sugars and polyols, organic acids and sweet amino acids, whereas a single-carbon source was beneficial to essential amino acid production, and a mixed-carbon source was beneficial to umami 5′-nucleotides and high mushroom production yield. High C/N values were beneficial to trehalose, arabitol, malic acid, and succinic acid production, while low C/N values were beneficial to mannitol and citric acid production. L. edodes fruiting bodies, harvested from a culture substrate containing single carbon, high proportion of cereal bran, had a more palatable taste quality, while a substrate containing bagasse and low C/N values was not suitable for L. edodes cultivation due to unfavourable taste quality. These results provide cultivation information how to obtain fruiting bodies L. edodes with more taste components. more...

It has been reported that phosphorus deprivation can induce β-carotene and triacylglycerol accumulation in Dunaliella salina cells. In this study, we aimed to elucidate the metabolic responses of D. salina to phosphorus deprivation, using gas chromatography-mass spectrometry as analytical tool. A total of 79 metabolites were identified in cells cultured in either phosphorus-deprived or replete media, including 18 amino acids, 28 other acids, 16 sugars, 12 alcohols, and 5 amino compounds. Hierarchical clustering was used to sort these metabolites into three groups with different change trends. Most amino acids and sugars, including the abiotic stress-related metabolites lysine, proline, trehalose, talose, and tagatose, increased, whereas N,N-dimethylglycine, L-serine, D-erythro-pentose, and D-ribose remained constant upon phosphorus deprivation. Multivariate statistical partial least squares and principal component analyses indicated that metabolite profiles were significantly changed upon phosphorus deprivation, and 18 biomarkers which can be used to distinguish the two culture conditions were identified. Stress-related polyamines such as cadaverine, antioxidants such as L-ascorbic acid, and L-methionine, as well as the osmolytes proline, mannitol, and arabitol, also increased. Furthermore, phosphorus deprivation resulted in increases of both saturated and unsaturated fatty acids in D. salina cells. These results suggest that phosphorus deprivation triggers comprehensive metabolic responses in D. salina which may be useful for future bioprocesses. more...

Arabitol is a polyalcohol which has about 70% of the sweetness of sucrose and an energy density of 0.2 kcal/g. Similarly to xylitol, it can be used in the food and pharmaceutical industries as a natural sweetener, a texturing agent, a dental caries reducer, and a humectant. Biotechnological production of arabitol from sugars represents an interesting alternative to chemical production. The yeast Scheffersomyces shehatae strain 20BM-3 isolated from rotten wood was screened for its ability to produce arabitol from L-arabinose, glucose, and xylose. This isolate, cultured at 28°C and 150 rpm, secreted 4.03 ± 0.00 to 7.97 ± 0.67 g/l of arabitol from 17–30 g/l of L-arabinose assimilated from a medium containing 20–80 g/l of this pentose with yields of 0.24 ± 0.00 to 0.36 ± 0.02 g/g. An optimization study demonstrated that pH 4.0, 32°C, and a shaking frequency of 150 rpm were the optimum conditions for arabitol production by the investigated strain. Under these conditions, strain 20BM-3 produced 6.2 ± 0.17 g/l of arabitol from 17.5 g/l of arabinose after 4 days with a yield of 0.35 ± 0.01 g/g. This strain also produced arabitol from glucose, giving much lower yields, but did not produce it from xylose. The new strain can be successfully used for arabitol production from abundantly available sugars found in plant biomass. more...

Lichens can be used as cost-effective biomonitors of elements from road traffic in the urban environment. However, the nature of comprehensive interactions between hazardous heavy metals from road traffic and lichen metabolites remains unclear. In this study, elemental and metabolic profiles of the lichen Dirinaria picta after exposure to road traffic for 3 months in 9 sites of Singapore city were investigated. Concentrations of 34 elements reveal strong correlations among each other and a general increase with traffic exposure level. A variety of metabolites, i.e. sugar alcohols, sugars, amino acids, low-molecular mass organic acids, secondary metabolites and nucleosides, are identified through a multi-platform approach combining liquid chromatography quadrupole time-of-flight mass spectrometry (LC-QTOF/MS), gas chromatography quadrupole mass spectrometry (GC-Q/MS) and 1H nuclear magnetic resonance (1H NMR). The total amount of the heavy metals Fe, Zn, Cu, Cd, Pb and Co can be significantly correlated with 21 metabolites positively and 9 metabolites negatively. These correlations reveal a heavy metal detoxification mechanism through ROS scavenging, osmoregulation and chelation with N atoms and sulfhydryl groups, as well as depletion of arabitol and inhibited synthesis of certain secondary metabolites such as quinones and steroids due to heavy metal stress. This study provides new insights into the metabolic toxicity and detoxification mechanisms in lichens under heavy metal exposure. more...

Liamocins are polyol lipids produced by the fungus Aureobasidium pullulans, and have selective antibacterial activity against Streptococcus species. Liamocins produced by A. pullulans strain NRRL 50380 on sucrose medium have a d-mannitol head group ester-linked to 3,5-dihydroxydecanoate acyl chains, three or four of which are joined together by 1,5-polyester bonds (liamocins Man-A1 and Man-B1), and similar 3'-O-acetylated analogs (Man-A2 and Man-B2). However, other types of liamocins are produced depending on the choice of strain and growth conditions. In the current study, growth on different polyols, but not sugars, resulted in considerable structural variation, including liamocins with d-galactitol (dulcitol), d-sorbitol (glucitol), d- and l-arabitol, d-xylitol, l-threitol and glycerol head groups. The head groups of liamocins produced on arabitol were shown to be entirely composed of d-arabitol. These liamocin variants were structurally characterized by NMR and MS, and tested for antibacterial activity. The new liamocin variants also had selective activity against Streptococcus. Liamocin structural variants are novel antibacterials against Streptococcus sp. that merit further investigation. more...

Lignocellulosic biomass of Japanese cedar, eucalyptus, bagasse, empty fruit bunch, and rice straw, has been directly converted into sugar alcohols such as sorbitol, mannitol, galactitol, xylitol, and arabitol, by supported platinum catalysts with hydrogen in water without any acid catalysts. Ball-milling pulverization of the lignocellulosic biomass increased the amount of total sugar alcohols from the direct conversion. The supported ruthenium and platinum bimetallic catalysts showed higher activity for the conversion of the lignocellulosic biomass. The amount of total sugar alcohols was increased up to 0.551 g from 1 g of milled bagasse by using the supported ruthenium and platinum bimetallic catalyst. more...

An elevated concentration of d-arabitol in urine, especially compared to that of l-arabitol or creatinine, is indicative of a fungal infection. For that purpose, we devised, fabricated, and tested chemical sensors determining d-arabitol. These chemosensors comprised the quartz crystal resonator (QCR) or extended-gate field-effect transistor (EG-FET) transducers integrated with molecularly imprinted polymer (MIP) film recognition units. To this end, we successfully applied a covalent approach to molecular imprinting, which involved formation of weak reversible covalent bonds between vicinal hydroxyl groups of arabitol and boronic acid substituents of the bithiophene functional monomer used. The MIP films were synthesized and simultaneously deposited on gold electrodes of quartz crystal resonators (Au-QCRs) or Au-glass slides by oxidative potentiodynamic electropolymerization. With the QCR and EG-FET chemosensors, the d-arabitol concentration was determined under flow-injection analysis and stagnant-solution binding conditions, respectively. Selectivity with respect to common interferences, and l-arabitol in particular, of the devised chemosensors was superior. Limits of detection and linear dynamic concentration ranges of the QCR and EG-FET chemosensors were 0.15 mM and 0.15 to 1.25 mM as well as 0.12 mM and 0.12 to 1.00 mM, respectively, being lower than the d-arabitol concentrations in urine of patients with invasive candidiasis (>220 μM). Therefore, the devised chemosensors are suitable for early diagnosis of fungal infections caused by Candida sp. yeasts. more...

Carbohydrates (such as levoglucosan) are a class of important water-soluble organic compounds in atmosphere. In this study, a high-performance liquid chromatography tandem mass spectrometry (HPLC-MS/MS) was applied to characterize carbohydrates in aerosol particles. Since carbohydrate was a kind of compound with low response in mass spectrometry, the conventional HPLC-MS/MS method was not sensitive enough to determine it. When acetate acid was added into mobile phase as buffer, the transition of [M+CH3COO]−→[M-H]− could be selected as the quantification ions. In the range from 1.0 μg L−1 to 20 μg mL−1, the coefficients of regression (r2) were more than 0.990, and relative standard deviations (RSD) for replicated injections were lower than 2%. The limit of detection (LOD) and quantification (LOQ) were lower than 2.5 ng L−1 and 10 ng L−1, respectively. The precision and accuracy were examined by spiked samples at three different concentration levels (10 μg L−1, 100 μg L−1, and 500 μg L−1) in five replicates. Recovery ratios ranged from 85% to 115% with RSD lower than 16%. Matrix effects of different carbohydrates ranged from 62% to 120%. The most sensitive HPLC-MS/MS method was developed and validated to analyze 40 aerosol samples successfully. The carbohydrates including three sugar alcohols (threitol, arabitol and sorbitol), one monosaccharide sugar (inositol), two disaccharides (sucrose, trehalose), one anhydrosugar (levoglucosan) and one 2-methyltetrols (2-Methylbutane-1,2,3,4-tretraol) were successfully quantified. more...

A growing number of studies are using specific primary sugar species, such as sugar alcohols or primary saccharides, as marker compounds to characterize and apportion primary biogenic organic aerosols (PBOAs) in the atmosphere. To better understand their annual cycles, as well as their spatiotemporal abundance in terms of concentrations and sources, we conducted a large study focusing on three major atmospheric primary sugar compounds (i.e., arabitol, mannitol, and glucose) measured in various environmental conditions for about 5300 filter samples collected at 28 sites in France. Our results show significant atmospheric concentrations of polyols (defined here as the sum of arabitol and mannitol) and glucose at each sampling location, highlighting their ubiquity. Results also confirm that polyols and glucose are mainly associated with the coarse rather than the fine aerosol mode. At nearly all sites, atmospheric concentrations of polyols and glucose display a well-marked seasonal pattern, with maximum concentrations from late spring to early autumn, followed by an abrupt decrease in late autumn, and a minimum concentration during wintertime. Such seasonal patterns support biogenic emissions associated with higher biological metabolic activities (sporulation, growth, etc.) during warmer periods. Results from a previous comprehensive study using positive matrix factorization (PMF) based on an extended aerosol chemical composition dataset of up to 130 species for 16 of the same sample series have also been used in the present work. The polyols-to-PMPBOA ratio is 0.024±0.010 on average for all sites, with no clear distinction between traffic, urban, or rural typology. Overall, even if the exact origin of the PBOA source is still under investigation, it appears to be an important source of particulate matter (PM), especially during summertime. Results also show that PBOAs are significant sources of total organic matter (OM) in PM10 (13±4 % on a yearly average, and up to 40 % in some environments in summer) at most of the investigated sites. The mean PBOA chemical profile is clearly dominated by contribution from OM (78±9 % of the mass of the PBOA PMF on average), and only a minor contribution from the dust class (3±4 %), suggesting that ambient polyols are most likely associated with biological particle emissions (e.g., active spore discharge) rather than soil dust resuspension. more...

Saccharomycopsis fibuligera KJJ81 isolated from nuruk is an amylolytic yeast that is widely used as a microbial starter in various fermented foods. Volatile and nonvolatile metabolites of S. fibuligera KJJ81 were investigated according to different carbon sources and cultivation times using a nontargeted metabolomic approach. Partial-least-squares discriminant analysis was applied to determine the major metabolites, which were found to be closely related to the clustering and discrimination of S. fibuligera KJJ81 samples. Some volatile metabolites derived from phenylalanine, such as 2-phenylethanol, 2-phenylethyl acetate, and ethyl phenylacetate, were predominantly found in cultivation medium containing glucose (YPD medium). In addition, the level of 2-phenylethanol increased continuously with the cultivation time. In terms of nonvolatile metabolites, carbohydrates (mannose, arabitol, and mannitol), fatty acids (palmitic acid and stearic acid), organic acids (oxalic acid and succinic acid), and amino acids (isoleucine, serine, alanine, glutamic acid, glycine, proline, phenylalanine, and threonine) were the main contributors to S. fibuligera KJJ81 samples cultivated in YPD medium according to cultivation time. These results show that the formation of volatile and nonvolatile metabolites of S. fibuligera KJJ81 can be significantly affected by both the carbon sources and the cultivation time. more...

The characteristics of biogenic aerosols in an urban area were explored by determining the composition and temporal distribution of saccharides in PM2.5 in Shanghai. The total saccharides showed a wide range of 9.4 ng/m3 to 1652.9 ng/m3, with the averaged concentrations of 133.1 ng/m3, 267.5 ng/m3, 265.1 ng/m3, and 674.4 ng/m3 in spring, summer, autumn, and winter, respectively. The saccharides include anhydrosaccharides (levoglucosan and mannosan), which were higher in cold seasons due to the increased biomass burning, saccharide alcohols (mannitol, arabitol, sorbitol), and monosaccharides (fructose, glucose), which were more abundant in warm seasons and attributed to the biological emissions. Through positive matrix factorization (PMF) analysis, four emission sources of saccharides were resolved, including biomass burning, fungal spores, plant decomposition, and pollen. Moreover, the process analysis of high concentrations of leveglucosan was conducted by backward trajectory and fire points. We found that concentrations of anhydrosaccharides were relatively stable under different pollution levels, while saccharide alcohols exhibited an obvious decrease with the concentration of PM2.5, indicating that biomass burning was not the core reason for heavy haze pollution. However, high level PM2.5 pollution might inhibit the effects of biological activities. more...

An array of ultra-long-chain fatty acid sugar alcohol monoesters, with behenic acid as an acyl representative and sugar alcohols altered from ethylene glycol to glycerol, erythritol, pentaerythritol, arabitol, xylitol and sorbitol, were enzymatically synthesized in high purity and selectivity. The molecular structures of the synthetic compounds were confirmed by 1H NMR, FT-IR and MS analysis, and the thermal properties were primarily characterized by DSC analysis. The molecular packing and thermal properties of synthetic sugar alcohol monobehenates (SAMBs) were investigated by Temp-Ramp-FT-IR. For in vivo application purposes, the enzymatic lipolysis of synthesized SAMBs was examined by a PPL (porcine pancreatic lipase)-mediated in vitro digestion test, and improved resistance of most SAMBs to enzymatic lipolysis, in comparison to glycerol monopalmitate, was observed. FT-IR spectroscopic analysis of the thermotropic phase transitions of the synthetic SAMBs indicated that the thermal collapse temperatures do not vary significantly as the polar head alters, suggesting their thermostabilities are largely governed by hydrophobic interactions among the alkanyl chains, while the size, properties and volume of the polar heads may determine the packing patterns. Systemic mapping of the structure–property–function relationship of SAMBs revealed the potential of these compounds for multipurpose applications. Ethylene glycol and glycerol monobehenates enable orthorhombic packing, and could find applications in cosmetic formulation, whereas sorbitol monobehenate is capable of forming stable surfactant-free nanoparticles, which could be excellent excipients for solid lipid nanoparticles for use as delivery cargo for drugs and food ingredients. more...

Concentration and optical characteristics of dissolved organic matter were studied in rainwater in the urban/coastal city of Thessaloniki, Northern Greece for 2-yr sampling period (2014-2016). The concentration of DOC ranged from 0.33-24.5mg/L with higher values measured in spring-summer period. Higher aromaticity and fluorescence intensity was observed in winter. Chromophoric organic matter represents a significant fraction of DOC that is highly correlated with fluorescence during cold period. Three factor spectral profiles of fluorescence were elucidated, with peaks at protein-like and humic-like area at different intensities. Fluorophores at shorter wavelengths are more susceptible to changes. DOC showed negative relationship with precipitation height, particularly during autumn and spring suggesting washout effect. NMR spectra showed the dominance of aliphatic protons in rainwater. Levoglucosan, sucrose and arabitol were determined in rainwater at concentrations more...

Biosurfactants have several desirable characteristics in the industrial sector: detergency, antimicrobial effects, skin hydration, and emulsibility. Several yeast glycolipids are currently being utilized in these capacities: sophorolipids, ustilagic acid, and mannosylerythritol lipids (MELs). An emerging class of glycolipids, termed polyol esters of fatty acids (PEFA), have recently been reported for Rhodotorula babjevae, a basidiomycetous yeast species that secretes hyperacetylated congeners of PEFA (typically with 3-6 acetylation modifications). While screening Rhodotorula species for surfactant production, we identified a new environmental isolate identified as Rhodotorula taiwanensis MD1149 that dropped the surface tension of the liquid medium, indicating that it produced a potent biosurfactant. Acid depolymerization of the purified biosurfactants, followed by gas chromatography-mass spectrometry (GC-MS) analysis revealed that the biosurfactants were composed of PEFA compounds composed mainly of mannitol and arabitol esters of 3-hydroxy fatty acid, 3-methoxy fatty acid, and fatty acids with a single double bond; chain lengths were mainly C16 and C18. Liquid chromatography-mass spectrometry (LC-MS) confirmed the predicted accurate mass of these compounds. Interestingly, PEFA compounds produced by Rhodotorula taiwanensis MD1149 were more surface active due to their hypoacetylation profile (0-4 acetylation modifications) compared to Rhodotorula babjevae MD1169. These disparate surface active properties, based on acetylation, change the hydrophilic-lipophilic balance (HLB) of these compounds, and their potential utility within industrial applications. more...

Carbohydrates have been widely reported in atmospheric aerosols, but have not previously been quantified in rainwater. We have identified and quantified a series of 11 specific compounds including monosaccharides (glucose, fructose, arabinose, galactose and pinitol), disaccharides (sucrose and trehalose), sugar alcohols (arabitol, dulcitol and mannitol) and the anhydrosaccharide levoglucosan. Rainwater analyzed in this study includes 52 distinct precipitation events in Wilmington, NC between June 2011 and October 2012. Our analysis indicates carbohydrates typically contribute more...

Low-molecular weight organic compounds (LMWOC) play a key role in soil respiration. Thus, a detailed understanding of their dynamics is important for any attempt to describe carbon cycling of ecosystems. Measurements of LMWOC have been compromised by the perturbation associated with traditional soil extraction methods. Due to the fast turnover of LMWOC, there are potential losses of compounds caused by ex situ methods. These losses should be reduced by microdialysis, a novel soil sampling technique that samples the delivery of compounds through induced diffusive flux rates (DFR), representing what an active organism is exposed to in situ, in the undisturbed soil. The present study used microdialysis to measure the DFR of LMWOC in the O-horizon of a boreal forest soil in northern Sweden at four occasions during the middle to end of the growing season. LMWOC was analyzed by untargeted metabolomic profiling, which identified 50 compounds, and targeted mass spectrometry, which quantified seasonal shifts in the abundance of 36 amino acids, organic acids and sugars. The amino acid proportion of the DFR decreased by 85% during the growing season. However, because the decrease in amino acids was compensated for by increases in organic acid and sugar fluxes, the total DFR remained approximately constant during the season. The organic acid pool was dominated by lactate (80%) in the middle of the season and consisted mainly of oxalate (40%) during the later season. A lower amount and more diverse composition of sugars were found compared to previous studies. Six sugars and polyols were detected, with mannitol, arabitol and glucose being the most abundant. Microdialysis has previously been explored to monitor amino acids and inorganic compounds, but the present research demonstrates the potential of the in situ microdialysis sampling when combined with metabolomics. This broadens the application of microdialysis to describing quantitative and qualitative changes in the dynamic composition of the DFRs in undisturbed soil and a similar approach may be used in future studies to study the actual soil processes governing soil LMWOC. more...

Sugar alcohols have been widely applied in the field of food and medicine for their unique properties. Compared to chemical production, microbial production of sugar alcohol has become attractive for its environmental and sustainable pattern. In this study, a potential yeast isolated from soil of Beijing suburbs was identified as Pichia anomala TIB-x229, and its key enzyme of d-arabitol dehydrogenase for microbial production of sugar alcohols was functionally characterized. This yeast could simultaneously produce d-arabitol, xylitol, and/or ribitol from a different ratio of sugar substrates at a high efficiency by bioconversion, and no glucose repression happened when mixed sugars of xylose and glucose were used as the substrates during the bioconversion. This yeast could also efficiently convert complicated feedstock such as xylose mother liquor to d-arabitol, xylitol, and ribitol with 55 % yields. To elucidate the conversion relationship of the sugar alcohols, especially d-arabitol and xylitol, the key d-arabitol dehydrogenase gene from P. anomala was cloned, expressed and purified for further in vitro characterization. The results showed that this d-arabitol dehydrogenase could catalyze arabitol to xylulose further, which is significant for xylitol production from glucose. Our study laid the foundation for improving the production of sugar alcohols by metabolic and fermentation engineering strategies. more...

The fermentation process for arabitol production from glycerol was developed using a Debaryomyces hansenii strain recently selected from a broad screening. The high-producing strain produced arabitol as the only detectable polyol from glycerol. In this work, the pH, dissolved oxygen concentration (DO), inoculum size and magnesium concentration, and the nitrogen-to-phosphorus (N/P) ratio were systematically evaluated for effects on cell growth rate and arabitol productivity. Among those evaluated, the medium with N/P = 9, DO of 5% air saturation and pH 3.5 supported the highest arabitol production. Under these optimal conditions, arabitol production of 40 g/L was achieved in 5 days compared to earlier studies with 15 g/L arabitol in 5 days. Volumetric productivity and specific productivity were successfully improved from 0.13 to 0.33 g/L-h and 0.007 to 0.02 g/g-h respectively with arabitol yield of 55% from glycerol. more...

The fungus Fusarium sp. isolated from saline soil was identified by the ITS1-5.8S-ITS2 and the D1/D2 domains of LSU RNA as a member of the Fusarium incarnatum-equiseti species group. Its growth patterns on media with different NaCl concentrations indicated its adaptation as halotolerance. The mechanisms of halotolerance included accumulation of arabitol (a five-atom noncyclic polyol), a decreased sterols/phospholipids ratio, elevated level of phosphatidic acids in the phospholipids, and increased unsaturation of phospholipids, which was especially pronounced in the idiophase. The mechanisms of halotolerance of the mycelial fungus Fusarium sp. are discussed in comparison with yeasts and yeastlike fungi. more...

Two classes of polar organic compounds, dicarboxylic acids (DCAs) and sugars/sugar anhydrides (S/SAs), were measured in airborne particulate matter in the area of Thessaloniki, northern Greece. The target compounds were measured simultaneously in two particle fractions PM10 and PM2.5 during cold and warm periods by employing extraction in an ultrasonic bath with a mixture of MeOH/DCM (1:2 v/v), derivatization with BSTFA-TMCS and GC-MS for analysis. At both fractions, phthalic was the predominant carboxylic acid during cold season and a-ketoglutaric acid in warm season, followed by maleic and malic. Levoglucosan was the dominant sugar anhydride during the cold and arabitol during the warm season. In total, the distribution of DCAs seemed to favor the PM2.5 particle fraction, probably due to anthropogenic emissions and photochemical formation. The relative contribution of DCAs to PM2.5 fraction was 0.9–3.2% in cold and 0.9–7.0% in warm period. Regarding S/SAs, levoglucosan was also predominantly distributed in fine particles, with relative contribution to this fraction 0.1–6.3% in cold and more...

Xylitol dehydrogenase (XDHA) and L-arabitol dehydrogenase (LADA) are two key enzymes in xylan metabolism catalyzing the oxidation of xylitol to D-xylulose and arabitol to L-xylulose, respectively. In Aspergillus oryzae, XDHA and LADA are encoded by xdhA and ladA. We deleted xdhA and ladA and xdhA-ladA to generate mutants with decreased dehydrogenase activities and increased xylitol production. The mutants were constructed by homologous transformation into A. oryzae P4 (∆pyrG) using pyrG as a selectable marker. The xylitol productivity of the mutants was measured using D-xylose as the sole carbohydrate source. xdhA, ladA, and the double-deletion mutant produced, respectively, 12.4 g xylitol/l with a yield of 0.24 g/g D-xylose, 12.4 g/l with a yield of 0.33 g/g D-xylose, and 8.6 g/l at a yield of 0.26 g/g D-xylose. more...

L-arabitol is used in the food and pharmaceutical industries. It can be secreted by genetically modified Saccharomyces cerevisiae carrying the genes responsible for pentose metabolism in yeast cells. The process of the biotransformation of L-arabinose to arabitol is highly dependent on culture conditions. The aim of this investigation was to use statistical response surface methodology (RSM) for optimization of biotransformation of L-arabinose to arabitol by a karyoductant of S. cerevisiae V30 and Pichia stipitis CCY 39501, named SP-K7. Batch cultures of yeast were performed according to a Plackett-Burman design, and three factors, rotation speed, L-arabinose concentration, and temperature, were chosen for a central composite design (CCD) applied in order to optimize the production of the polyol by the karyoductant. On the basis of results obtained using 20 combinations of batch cultures of karyoductant SP-K7, the optimal levels of the factors were determined as: rotation speed 150 rpm, concentration of L-arabinose 32.5 g/l, and temperature 28 degrees C. In such conditions, the predicted concentration of arabitol after two days of incubation of SP-K7 should be 18.367 g/l. The value of R2 = 0.93195 suggested that this model was well-fitted to the experimental data. A verification of the model in experimental conditions confirmed its usefulness. more...

A method of detecting carbohydrates (fucose, trehalose, mannitol, arabitol, mannose, glucose, galactose, fructose, and ribose) by high-performance anion chromatography-pulsed amperometric detection (HAPEC-PAD) was established. The conditions are: CarboPac MA1 column, NaOH as the eluent, temperature 30°C, Au working electrode, Ag/AgCl reference electrode, and flow rate 0.4 mL/min. These nine analytes, which yielded high resolution by this method, could be detected in 40 minutes. Mushrooms were tested and good precision, stability, and reproducibility were achieved. This method is suitable for mushroom samples and could support research and development on sugar and sugar alcohol, which contains special effects. more...

The saprophytic fungus Hypocrea jecorina (anamorph, Trichoderma reesei ) is an important native producer of hydrolytic enzymes, including xylanases. Regarding principles of sustainability, cheap and renewable raw materials, such as d -xylose (the backbone monomer of xylan), have been receiving increasing attention from industries. Recently, it was demonstrated that small (0.5 to 1 mM) amounts of d -xylose induce the highest expression of xylanase in H. jecorina . However, it was also reported that active metabolism of d -xylose is necessary for induction. In this report, we demonstrate that xylitol, the next intermediate in the pentose pathway after d -xylose, does not trigger transcription of xylanase-encoding genes in H. jecorina QM9414. The highest level of transcription of xylanolytic enzyme-encoding genes occurred in an xdh1 (encoding a xylitol dehydrogenase) deletion strain cultured in the presence of 0.5 mM d -xylose, suggesting that a metabolite upstream of xylitol is the inducer. The expression levels of xylanases in an xdh1 - lad1 double-deletion strain were lower than that of an xdh1 deletion strain. This observation suggested that l -xylulose is not an inducer and led to the hypothesis that l -arabitol is the actual inducer of xylanase expression. A direct comparison of transcript levels following the incubation of the H. jecorina parental strain with various metabolites of the pentose pathway confirmed this hypothesis. In addition, we demonstrate that xyr1 , the activator gene, is not induced in the presence of pentose sugars and polyols, regardless of the concentration used; instead, we observed low constitutive expression of xyr1 . more...

Glycerol is a major by-product from biodiesel production, and developing new uses for glycerol is imperative to overall economics and sustainability of the biodiesel industry. With the aim of producing xylitol and/or arabitol as the value-added products from glycerol, 214 yeast strains, many osmotolerant, were first screened in this study. No strains were found to produce large amounts of xylitol as the dominant metabolite. Some produced polyol mixtures that might present difficulties to downstream separation and purification. Several Debaryomyces hansenii strains produced arabitol as the predominant metabolite with high yields, and D. hansenii strain SBP-1 (NRRL Y-7483) was chosen for further study on the effects of several growth conditions. The optimal temperature was found to be 30°C. Very low dissolved oxygen concentrations or anaerobic conditions inhibited polyol yields. Arabitol yield improved with increasing initial glycerol concentrations, reaching approximately 50% (w/w) with 150 g/L initial glycerol. However, the osmotic stress created by high salt concentrations (≥50 g/L) negatively affected arabitol production. Addition of glucose and xylose improved arabitol production while addition of sorbitol reduced production. Results from this work show that arabitol is a promising value-added product from glycerol using D. hansenii SBP-1 as the producing strain. more...

The genes encoding xylitol dehydrogenase (Texdh) and L: -arabitol dehydrogenase (Telad) are involved in the fungal pentose pathway and were isolated from the thermophilic fungus Talaromyces emersonii, expressed in Escherichia coli, and the products purified to homogeneity. TeXDH showed activity toward xylitol and D: -sorbitol. TeLAD was active with L: -arabitol, xylitol, and D: -sorbitol. Phylogenetic analysis showed TeLAD has evolved from D: -sorbitol dehydrogenase as a result of environmental adaptation. Substrate specificity studies indicate that TeXDH is likely to have evolved from the more broadly acting TeLAD. Texdh and Telad expression was inducible by the same carbon sources responsible for induction of genes involved in biomass degradation, suggesting for the first time a coordinated regulatory control mechanism for expression of genes encoding extracellular hydrolases and intracellular metabolic genes in the pentose utilization pathways of T. emersonii. These data also suggest that TeXDH and TeLAD may be valuable in the production of xylitol, L: -arabitol, and ethanol from renewable resources rich in pentose sugars. more...

An improved high-performance anion-exchange chromatography (HPAEC) with pulsed amperometric detection (PAD) method is developed and validated for simultaneous determination of atmospherically relevant sugar alcohols, monosaccharides, and monosaccharide anhydrides. The improved method enables the separation of levoglucosan and arabitol which were not or insufficiently separated by the previous HPAEC–PAD methods. Reproducibility of the method was tested for both standard solutions and atmospheric aerosol samples. The peak area relative standard deviation (RSD%) of standard solutions were found to be lower than 1.5% for consecutive analyses (n ¼ 3) and lower than 4% for day to day variation (n ¼ 9). The peak area RSD% of atmospheric samples with typical European wintertime monosaccharide concentrations (n ¼ 9) was found to be similar to that of standard solutions. Limits of detection ranged from 0.002 mg L �1 for inositol to 0.08 mg L �1 for fructose. The developed method offers more...

Metabolomics is a rapidly growing branch of post-genomic chemical biology. The fission yeast Schizosaccharomyces pombe is an excellent eukaryotic model organism. Although the entire S. pombe genome has been sequenced and detailed transcriptomic analyses were performed, little metabolic profiling has been done. Here we report the first global semi-quantitative analysis of the S. pombe metabolome using liquid chromatography high-resolution mass spectrometry. Procedures to obtain metabolic compounds from S. pombe extracts were established. One hundred and twenty-three distinct metabolites were identified while approximately 1900 peaks from the approximately 6000 observed were assigned. A software system (MZviewer) was developed to visualize semi-quantitative metabolome data using a dynamically generated scatter plot. We examined the metabolome of S. pombe cells exponentially grown in synthetic culture medium (EMM2) at two different temperatures, 26 degrees C and 36 degrees C. The profiles were similar except for varying amounts of certain amino acids and a significant increase in several compounds at 36 degrees C, such as trehalose (200-fold), glycerophosphoethanolamine (50-fold), arabitol (16-fold), ribulose (8-fold), and ophthalmic acid (5-fold). Reproducibility was demonstrated using a deletion mutant sib1Delta that lacked ferrichrome synthetase and showed no significant metabolic effects except the disappearance of the hexapeptide ferrichrome and the appearance of a putative dipeptide precursor. Taking advantage of the metabolic profile similarity at 26 degrees C and 36 degrees C, we analyzed the metabolome of a temperature-sensitive hcs1-143 mutant defective in the HMG-CoA synthase. As expected, HMG-CoA was decreased. In addition, extensive secondary metabolic effects, including a decrease in urea cycle intermediates and an increase in acetylated compounds, were observed. These findings confirm that S. pombe can be applied as an appropriate model to monitor metabolic responses to environmental conditions as well as genetic perturbations. more...

The monosaccharide and polyalcohol composition of 28-samples of different commercial tannins, including oak wood, grape seed and skin, plant gall, chestnut, quebracho and gambier, has been evaluated by gas chromatography–mass spectrometry after derivatization into their trimethylsilyl ethers. Quercitol was found to be characteristic of oak tannins, whereas gall plant tannins could be differentiated by their content of pinitol. Myo -inositol and arabitol were detected in tannins from quebracho. These polyalcohols, together with muco -inositol and chiro -inositol, were found in tannins from chestnut while bornesitol was found to be characteristic of tannins from gambier. Monosaccharide composition also helped to distinguish among tannin origins: arabinose, xylose, fructose and glucose were quantified in oak, quebracho and chestnut tannins, whereas only fructose and glucose were detected in plant gall and grape tannins. These results imply that the qualitative study of monosaccharides and polyalcohols could help to determine and control the authenticity of enological tannins. more...

A non-targeted metabolomics approach was used to identify significant changes in metabolism upon exposure of the wheat pathogen Stagonospora nodorum to 0.5 M NaCl. The polyol arabitol, and to a lesser extent glycerol, was found to accumulate in response to the osmotic stress treatment. Amino acid synthesis was strongly down-regulated whilst mannitol levels were unaffected. A reverse genetic approach was undertaken to dissect the role of arabitol metabolism during salt stress. Strains of S. nodorum lacking a gene encoding an l-arabitol dehydrogenase (abd1), a xylitol dehydrogenase (xdh1) and a double-mutant lacking both genes (abd1xdh1) were exposed to salt and the intracellular metabolites analysed. Arabitol levels were significantly up-regulated upon salt stress in the xdh1 strains but were significantly lower than the wild-type. Arabitol was not significantly different in either the abd1 or the abd1xdh1 strains during osmotic stress but the concentration of glycerol was significantly higher indicating a compensatory mechanism in operation. Genome sequence analysis identified a second possible enzyme capable of synthesizing arabitol explaining the basal level of arabitol present in the abd1xdh1 strains. This study identified that arabitol is the primary compatible solute in S. nodorum but in-built levels of redundancy are present allowing the fungus to tolerate osmotic stress. more...

Osmoprotectants stabilize proteins and membranes against the denaturing effect of high concentrations of salts and other harmful solutes. In yeast, arabitol dehydrogenase (ArDH) reduces D-ribulose to D-arabitol where D-ribulose is derived by dephosphorylating D-ribulose-5-PO4 in the oxidized pentose pathway. Osmotolerance in plants could be developed through metabolic engineering of chloroplast genome by introducing genes encoding polyols since chloroplasts offer high level transgene expression and containment. Here, we report that ArDH expression in tobacco chloroplasts confers tolerance to NaCl (up to 400 mM). Transgenic plants compared to wild type (WT) survived for only 4-5 weeks on 400 mM NaCl whereas plants remained green and grew normal on concentrations up to 350 mM NaCl. Further, a-week-old seedlings were also challenged with poly ethylene glycol (PEG, up to 6%) in the liquid medium, considering that membranes and proteins are protected under stress conditions due to accumulation of arabitol in chloroplasts. Seedlings were tolerant to 6% PEG, suggesting that ARDH enzyme maintains integrity of membranes in chloroplasts under drought conditions via metabolic engineering. Hence, the gene could be expressed in agronomic plants to withstand abiotic stresses. more...

To compare production of antibacterial liamocins (polyol lipids) by diverse strains of Aureobasidium pullulans grown on different culture media. Liamocins produced by strains of A. pullulans have potential agricultural and pharmaceutical applications as antibacterials with specificity against Streptococcus spp. Six strains of A. pullulans were characterized for liamocin production on four different culture media. The choice of strain and culture medium affected growth, liamocin yields, and production of contaminating pigments. Best growth and highest liamocin yields were obtained using A. pullulans strain NRRL 50384 grown on a sucrose basal medium. Unexpectedly, the choice of strain and culture medium also affected the structure of liamocins produced, providing novel types of liamocins. Liamocins varied not only in the ratios of trimer and tetramer polyester tail groups, but also in the nature of the polyol headgroup, which could include mannitol, arabitol, or glycerol. The ability to conveniently produce novel types of liamocins in good yields will provide novel antibacterials for applied uses, and facilitate structure-function studies on the mechanism of antibacterial activity. more...

Surface tension curves were measured using the Wilhelmy method for the monoacyl glycerols, erythritols and xylitols with the acyl chain lengths of 8, 10, 12 and 14 at 25, 32 and 39 °C. The curves were also measured for the monolauroyl esters of arabitol, ribitol and sorbitol. The critical micelle concentration, CMC, for a series of sugar alcohol esters with different acyl chains decreased as the chain length increased. The numbers and orientations of the hydroxyl groups of the sugar alcohols with the monolauroyl residue did not affect the CMC, while the surface tension at the CMC, γCMC, increased as the number of hydroxyl groups increased except for the monoacyl xylitols. The residual area per molecule was also independent of the types of hydrophilic and hydrophobic moieties, and was in the range of 0.3–0.4 nm2. The CMC values did not depend on the temperature for any of the tested monoacyl sugar alcohols, indicating that the enthalpy of the micellization was nearly zero. Because the free energy of the micellization was negative for every ester, the entropy of micellization was positive. more...

Bulk aerosols (>1 μm) were collected continuously above the canopy at the Howland Experimental Forest, Maine, USA from May to October 2002. Each sample integrated over an approximately 2-week period. Mono- and disaccharide sugars were extracted using a microscale technique and were analyzed as their TMS derivatives by gas chromatography–mass spectrometry (GC–MS). Concentrations of total aerosol sugars ranged from 10 to 180 ng m −3 . Glucose was the most abundant sugar (40–75% of the total sugars). The monosaccharides arabinose, fructose, galactose, mannose, arabitol and mannitol, and the disaccharides sucrose, maltose and mycose (aka trehalose) were also present in lower concentrations. The sugar composition in the aerosols varied seasonally. Fructose and sucrose were prevalent in early spring and decreased in relative abundance as the growing season progressed. Sugar polyols (arabitol and mannitol) and the disaccharide mycose (a fungal metabolite) were more prevalent in autumn during the period of leaf senescence. The changes in the sugar composition in the aerosol samples appear to reflect the seasonality of sugar production and utilization by the ecosystem. Plant waxes were present as significant components also indicating an input from biogenic background. Smoke plumes from Quebec forest fires passed over the Howland site in early July 2002. Levoglucosan, a biomarker of biomass burning, increased by an order of magnitude in the aerosol samples collected during this time. Glucose, mannose, arabinose, galactose, and also, plant waxes increased in concentration by factors of 2–5 in the smoke-impacted samples, indicating that wildfires enhance atmospheric emissions of uncombusted organic compounds. In contrast, concentrations of fructose, sugar polyols and disaccharides were not significantly higher in the smoke-impacted samples and indicated that biomass burning was not a significant source of these compounds in the aerosols. more...

Polyol production has been studied in Aspergillus niger under different conditions. Fermentations have been run using high concentration of glucose or xylose as carbon source and ammonium or nitrate as nitrogen source. The growth of biomass, as freely dispersed hyphae, led to an increase of medium viscosity and hereby a decrease in mass transfer, especially oxygen transfer. The consequence was a decrease in DOT and the occurrence of a switch between fully aerobic conditions and oxygen-limited conditions. Metabolite quantification showed that polyols were the main metabolic products formed and represented up to 22% of the carbon consumed in oxygen-limited conditions. The polyol concentration and the polyol pattern depended strongly on the environmental conditions. This is due to a complex regulation of polyol production and to the fact that each polyol can fulfill different functions. In this study, erythritol, xylitol, and arabitol were produced as carbon storage compounds when the flux through the PP pathway exceeded the need in ribulose-5-phosphate for the biomass synthesis. Glycerol, erythritol, and xylitol seem to be involved in osmoregulation. Mannitol was produced when the catabolic reduction of charge was high. Its production involves the enzyme NAD-dependent mannitol-1-phosphate dehydrogenase and seems to be the main cytosolic route for the NADH reoxidation during oxygen limitation. more...

The aim of this study was the production of high numbers of M. anisopliae blastospores with enhanced germination efficiency under conditions of water-stress (ecological fitness). Different nitrogen sources and concentrations were screened for their ability to induce blastospore production while keeping pH and water activity (a w) at fixed levels (6.8, and 0.98 a w, respectively). After optimum nitrogen status was determined (cornsteep solid (CS) + yeast extract (YE); or cottonseed flour (CF) + YE), the effect of interaction of nitrogen source, pH (3.5, 5, 6.8, 8, 9 and 10) and solutes for a w adjustment (KCl, NaCl, PEG 200) on blastospore production, endogenous polyol content (glycerol, erythritol, arabitol and mannitol) and total protein, were determined. For both ionic (NaCl, KCl) and non-ionic solutes (PEG 200), optimum blastospore production (between 4×107 and 2×108 blastospores ml−1) and growth occurred in the pH range 6.8–8, with the CF + YE nitrogen profile giving higher yields than CS + Y... more...