Inhibition of mycotoxin deoxynivalenol generation by using selenized glucose

Selenized glucose can be easily prepared via the selenization reaction of glucose using in situ generated NaHSe as the selenization reagent. The technique has been industrialized to produce the chemical in kilogram scale, making it an easily available material in laboratory presently. The selenized glucose may be widely used as the starting material for the preparation of selenium-containing catalysts, as the organoselenium additive for feeds, and as the efficient selenium-enriched foliar fertilizers. In this work, we found that treating Fusarium graminearum, a fungal pathogen inciting wheat scab disease, with selenium glucose could significantly inhibit the generation of the deoxynivalenol (DON) toxin, which might be a breakthrough for reducing the detriment of the wheat scab disease.     

High Solids Enzymatic Hydrolysis of Pretreated Lignocellulosic Materials with a Powerful Stirrer Concept

In this study, we present a powerful stirred tank reactor system that can efficiently hydrolyse lignocellulosic material at high solid content to produce hydrolysates with glucose concentration > 100 g/kg. As lignocellulosic substrates alkaline-pretreated wheat straw and organosolv-pretreated beech wood were used. The developed vertical reactor was equipped with a segmented helical stirrer, which was specially designed for high biomass hydrolysis. The stirrer was characterised according to mixing behaviour and power input. To minimise the cellulase dosage, a response surface plan was used. With the empirical relationship between glucose yield, cellulase loading and solid content, the minimal cellulase dosage was calculated to reach at least 70 % yield at high glucose and high substrate concentrations within 48 h. The optimisation resulted in a minimal enzyme dosage of 30 FPU/g dry matter (DM) for the hydrolysis of wheat straw and 20 FPU/g DM for the hydrolysis of beech wood. By transferring the hydrolysis reaction from shaking flasks to the stirred tank reactor, the glucose yields could be increased. Using the developed stirred tank reactor system, alkaline-pretreated wheat straw could be converted to 110 g/kg glucose (76 %) at a solid content of 20 % (w/w) after 48 h. Organosolv-pretreated beech wood could be efficiently hydrolysed even at 30 % (w/w) DM, giving 150 g/kg glucose (72 %).     

Xylanase production by Penicillium canescens 10–10c in solid-state fermentation

Filamentous fungi have been widely used to produce hydrolytic enzymes for industrial applications, including xylanases, whose levels in fungi are generally much higher than those in yeast and bacteria. We evaluated the influence of carbon sources, nitrogen sources, and moisture content on xylanase production by Penicillium canescens 10–10c in solid-state fermentation. Among agricultural wastes tested (wheat bran, untreated wheat straw, treated wheat straw, beet pulp, and soja meal), untreated wheat straw gave the highest production of xylanase. Optimal initial moisture content for xylanase production was 83%. The addition of 0.4 g of xylan or easily metabolizable sugar, such as glucose and xylose, at a concentration of 2% to wheat straw enhanced xylanase production. In solid-state fermentation, even at high concentrations of glucose or xylose (10%), catabolic repression was minimized compared to the effect observed in liquid culture. Yeast extract was the best nitrogen source among the nitrogen sources investigated: peptone, ammonium nitrate, sodium nitrate, ammonium chloride, and ammonium sulfate. A combination of yeast extract and peptone as nitrogen sources led to the best xylanase production.     

Investigations on Hydrolytic Activities from Stachybotrys microspora and Their Use as an Alternative in Yeast DNA Extraction

Stachybotrys microspora is a filamentous fungus characterized by the secretion of multiple hydrolytic activities (cellulolytic and non-cellulolytic enzymes). The production of these biocatalysts was studied under submerged culture using glucose, cellulose, and wheat bran as carbon sources. Endoglucanases, pectinases, xylanases, β-glucanases, chitinases, and proteases were induced on cellulose-based medium and repressed on glucose in both strains with higher amounts produced by the mutant. β-glucosidases were roughly equally produced by both strains under glucose and cellulose conditions. The yield of chitinases, β-glucanases, and proteases produced by Stachybotrys strains was as much higher than the commercialized lysing enzyme called “zymolyase,” currently used in yeast DNA extraction. In this context, we showed that S. microspora hydrolases can be successfully applied in the extraction of yeast DNA.     

Localized surface plasmon resonance sensing detection of glucose in the serum samples of diabetes sufferers based on the redox reaction of chlorauric acid

In this paper, the formation of gold nanoparticles (Au NPs) as a result of the thermo-active redox reaction of chlorauric acid (HAuCl₄) and glucose in alkaline medium was identified by measuring the plasmon resonance absorption, localized surface plasmon resonance (LSPR), and transmission electron microscopy (TEM) images, for the formation of Au NPs displays characteristic plasmon resonance absorption bands and corresponding LSPR signals. It was found that the resulted LSPR signals could be easily detected with a common spectrofluorometer. With increasing glucose concentration, the LSPR intensity displays linear response with the glucose content over the range from 2.0 to 250.0 μmol l⁻¹. Thus, a novel assay of glucose was established with the limits of determination (3σ) being 0.21 μmol l⁻¹, and the detection of glucose could be made easily in the serum samples of diabetes sufferers. Mechanism investigations showed that the activation energy and molar ratio of the reaction were 34.8 kJ mol⁻¹ and 3:2, respectively.     

Synthesis of selenium-doped carbon from glucose: An efficient antibacterial material against Xcc

Se/C as a novel carbon-based biomaterial was prepared by using cheap and abundant glucose as the carbon source. It was highly active and could well restrain Xanthomonas campestris pv. campestris (EC₅₀ = 4.7403 μg/mL), a very harmful germ causing the devastating cabbage black rot disease and resulting in huge economic losses. As a type of carbon material insoluble in water, Se/C is bio-compatible and can adhere onto the leaves of the plants to allow a slow and sustained release of its efficacy, affording an efficient method for the cabbage black rot disease prevention and cure. This work as the first report on the bioactivity studies of Se/C significantly expands the application scopes of the selenium-containing materials and may draw continuous attentions from a broad field.     

Production and application of amylases of Rhizopus oryzae and Rhizopus microsporus var. oligosporus from industrial waste in acquisition of glucose

Amylases from Rhizopus oryzae and Rhizopus microsporus var. oligosporus were obtained using agro-industrial wastes as substrates in submerged batch cultures. The enzymatic complex was partially characterised for use in the production of glucose syrup. Type II wheat flour proved better than cassava bagasse as sole carbon source for amylase production. The optimum fermentation condition for both microorganisms was 96 hours at 30°C and the amylase thus produced was used for starch hydrolysis. The product of the enzymatic hydrolysis indicated that the enzyme obtained was glucoamylase, only glucose as final product was attained for both microorganisms. R. oligosporus was of greater interest than R. oryzae for amylase production, taking into account enzyme activity, cultivation time, thermal stability and pH range. Glucose syrup was produced using concentrated enzyme and 100 g L^?1 starch in a 4 hours reaction at 50°C. The bioprocess studied can contribute to fungus glucoamylase production and application.     

Effect of Water and Ethanol Extracts from Hericium erinaceus Solid-State Fermented Wheat Product on the Protection and Repair of Brain Cells in Zebrafish Embryos

Extracts from Hericium erinaceus can cause neural cells to produce nerve growth factor (NGF) and protect against neuron death. The objective of this study was to evaluate the effects of ethanol and hot water extracts from H. erinaceus solid-state fermented wheat product on the brain cells of zebrafish embryos in both pre-dosing protection mode and post-dosing repair mode. The results showed that 1% ethanol could effectively promote zebrafish embryo brain cell death. Both 200 ppm of ethanol and water extracts from H. erinaceus solid-state fermented wheat product protected brain cells and significantly reduced the death of brain cells caused by 1% ethanol treatment in zebrafish. Moreover, the zebrafish embryos were immersed in 1% ethanol for 4 h to cause brain cell damage and were then transferred and soaked in the 200 ppm of ethanol and water extracts from H. erinaceus solid-state fermented wheat product to restore the brain cells damaged by the 1% ethanol. However, the 200 ppm extracts from the unfermented wheat medium had no protective and repairing effects. Moreover, 200 ppm of ethanol and water extracts from H. erinaceus fruiting body had less significant protective and restorative effects on the brain cells of zebrafish embryos. Both the ethanol and hot water extracts from H. erinaceus solid-state fermented wheat product could protect and repair the brain cells of zebrafish embryos damaged by 1% ethanol. Therefore, it has great potential as a raw material for neuroprotective health product.     

Production of Mycophenolic Acid by <Emphasis Type="Italic">Penicillium brevicompactum</Emphasis> Using Solid State Fermentation

Solid-state fermentation using the microfungus Penicillium brevicompactum for the production of mycophenolic acid is reported in this paper. Of the initial substrates tested (whole wheat, cracked wheat, long grain Basmati rice, and short grain Parmal rice), Parmal rice proved to be the best. Under initial conditions, using steamed Parmal rice with 80% (w/w) initial moisture content, a maximum mycophenolic acid concentration of 3.4 g/kg substrate was achieved in 12 days of fermentation at 25 °C. The above substrate was supplemented with the following additional nutrients (g/L packed substrate): glucose 40.0, peptone 54.0, KH₂PO₄ 8.0, MgSO4?7H₂O 2.0, glycine 7.0, and methionine 1.65 (initial pH 5.0). A small amount of a specified trace element solution was also added. The final mycophenolic acid concentration was increased to nearly 4 g/kg substrate by replacing glucose with molasses. Replacing Parmal rice with rice bran as substrate further improved the mycophenolic acid production to nearly 4.5 g/kg substrate.     

Scrutinizing the facile growth of β-Ag2Se fine films

Silver selenide thin film is one of the best candidates for thermoelectric devices. In the previous report, we demonstrated that high-performanced [201] oriented β-Ag₂Se thin films can be prepared by direct metal surface element reaction (DMSER) solution selenization in a really short time at room temperature. However, the underlying mechanism of the fast reaction process were not discussed in depth. Herein, based on hard soft acid base (HASB) theory and strong oxidation, we further explored the possible reaction mechanism of the in-situ growth of β-Ag₂Se thin films as the function of the reaction time. The time-dependent experimental results showed that the formation of the β-Ag₂Se on elemental Ag precursor (∼690 nm thick) in Se/Na₂S precursor solution is in a growth driven mode with no obvious orientation or growth rate selections to the elemental Ag precursors. Our investigations provide a prerequisite for the further preparation of thermoelectric materials with excellent properties.     

A Physiologically Regulated Multidomain Cystatin of Wheat Shows Stage-Dependent Immunity Against Karnal Bunt (Tilletia indica)

To identify novel components of basal resistance against the Tellitia indica of wheat, breeding for disease resistance was carried out on resistant and susceptible genotype of Karnal Bunt. The different members of wheat cystatin gene families were cloned, and their role in triggering differential resistance through co-expression was analyzed in our lab. The multidomain wheat cystatin (WCM) is a proteinase inhibitor characterized by cloning the gene from susceptible (WH542) and resistant genotype (HD 29). A WCM cDNA was isolated from both genotypes and sequenced. The WCM had a highly conserved N-terminal cystatin domain and a long C-terminal extension containing a second region, which exhibited similarity to the cystatin domain. The expression level was significantly (P?>?0.001) higher in resistant compared to susceptible genotype at all the physiological stages of wheat spikes. In order to characterize the biochemical properties of WCM, the coding sequence was expressed in Escherichia coli using pET expression vector. The recombinant WCM was purified from soluble fraction of the cell extract by using affinity chromatography. WCM, with 23 KDa molecular mass, showed cysteine proteinase inhibitory activity against papain (Ki 3.039?×?10^?7?M) as determined by using BAPNA as substrate. Furthermore, it was able to arrest the fungal mycelial growth of T. indica. Hyphae growth was inhibited, and morphological changes such as swelling and fragmentation of the fungus were observed. Overall, these observations suggest an endogenous high expression of cystatin, possibly associated with the resistance of wheat against Karnal bunt.     

Synthesis of High Quality CuInSe2 Films with Oxygen Precursor by Non-vacuum Process

The CuInSe₂ absorber was synthesized by non-vacuum process with a simple and low-cost method, which fabricated absorber layer of thin-film solar cell. The extra amount of Se was added into the ink to help reduction of the oxide and solid Se fountain was used to provide Se atmosphere during the selenization progress. The influence of same factors was investigated, such as the time of reduction in H₂, the time of selenization and the Se vapor pressure. The selenizaion, processed at 550 °C for 60 min with the Se vapor pressure at 1.90 kPa, resulted in high quality CuInSe₂ layer with very good chemical composition.     

Synthesis of (R)-propane-1,2-diol from lactides by dynamic kinetic resolution

The dynamic kinetic resolution (DKR) of rac-1-tert-butoxypropan-2-ol with isopropenyl acetate in the presence of Novozyme 435 and a ruthenium catalyst produces enantiomerically pure (R)-1-tert-butoxy-2-acetoxy-propane (>99.5 %ee) in a good yield. The product can be easily transformed into (R)-propane-1,2-diol without loss of stereoselectivity. Together with recently published procedures, the herein described DKR offers the possibility to use any lactide source as starting material for the production of (R)-propane-1,2-diol. The chiral diol may serve as the chiral building block for the synthesis of important enantiopure esters, like propylene carbonate, chiral polymers, etc.     

Effect of process production on antinutritional,nutrition, and physicochemical properties of modified sorghum flour

Increasing wheat flour consumption makes increasing of wheat import value in Indonesia. Wheat flour has good nutritions, sufficient gluten compound, high glucose and high glycemic index. Therefore, substituting wheat with sorghum (local crop) which has gluten free, a low glucose and glycemic index value, can overcome food security problem in Indonesia. However, it has a high phytic acid and tannin (antinutrients). This study focuses on the reduction of antinutrients by three processes (fermentation treatment using lactic acid bacterias (LAB), including Lactobacillus bulgaricuss, Lactobacillus casei, and Lactobacillus brevis (1); NaOH solution submersion (2); and combining NaOH solution submersion followed by fermentation (3)). It was found that combining NaOH solution submersion and fermentation resulted in reduction of antinutrient contents (phytic acid decreased from 11.9% to 0.117% and the tannin decreased from 6.16% to 0.063%) and improved nutrition composition (such as protein content increased from 8.59% to 14.67%) on sorghum so that sorghum could be feasible to be an alternative source of wheat substitution in flour production. Moreover, the glucose content and glycemic Index (GI) of modified sorghum flour was decreased from 2.75 to 1.83%, and decreased from 40.79 to 36.33, respectively.     

Fermentation of Biologically Pretreated Wheat Straw for Ethanol Production: Comparison of Fermentative Microorganisms and Process Configurations

The pretreatment of lignocellulosic biomass with white-rot fungi to produce bioethanol is an environmentally friendly alternative to the commonly used physico-chemical processes. After biological pretreatment, a solid substrate composed of cellulose, hemicellulose and lignin, the two latter with a composition lower than that of the initial substrate, is obtained. In this study, six microorganisms and four process configurations were utilised to ferment a hydrolysate obtained from wheat straw pretreated with the white-rot fungus Irpex lacteus. To enhance total sugars utilisation, five of these microorganisms are able to metabolise, in addition to glucose, most of the pentoses obtained after the hydrolysis of wheat straw by the application of a mixture of hemicellulolytic and cellulolytic enzymes. The highest overall ethanol yield was obtained with the yeast Pachysolen tannophilus. Its application in combination with the best process configuration yielded 163 mg ethanol per gram of raw wheat straw, which was between 23 and 35 % greater than the yields typically obtained with a conventional bioethanol process, in which wheat straw is pretreated using steam explosion and fermented with the yeast Saccharomyces cerevisiae.     

Mixed ferrocene-glucose oxidase-carbon-paste electrode for amperometric determination of glucose

The simultaneous incorporation of ferrocene derivatives and glucose oxidase into a carbon-paste matrix results in an effective microelectrode for sensing glucose. The close proximity of the enzymatic, redox mediating and sensing sites offers extremely short response times (t_95%=18 s) compared with early ferrocene-based glucose sensors. Several ferrocene derivatives are evaluated, with 1,1′-dimethylferrocene yielding the best results. The influence of the paste composition, operating potential, glucose concentration and other variables is described. The incorporation of stearic acid into the enzyme-containing paste greatly reduces the interference due to ascorbic acid. The microelectrode is easily fabricated, as time-consuming multi-step immobilization schemes are eliminated. Flow-injection measurements at a rate of 90 samples per hour and relative standard deviations of 3% are also reported.     

A short and efficient synthesis of (R)-(−)-sporochnol A

A short and efficient synthesis of (R)-(−)-sporochnol A in five steps and 9% overall yield has been developed. The sequence uses as starting material the easily available enantiopure monoketal derived from 1,4-cyclohexanedione and (R,R)-hydrobenzoin that serves as a chiral auxiliary.     

Application of a magnetic immobilizedβ-glucosidase in the enzymatic saccharification of steam-exploded lignocellulosic residues

Aβ-glucosidase preparation derived fromAspergillus niger was immobilized onto a magnetic support and used in the enzymatic saccharification of a lignocellulosic material. The enzyme was immobilized onto polyethyleneimine-glutaraldehyde activated magnetite (PAM) and also onto titanium (IV) oxide (TiO₂)-coated magnetite (TAM). Although > 80% of the protein applied was immobilized, only 15–27% of the enzyme activity was recovered after immobilization. Theβ-glucosidase immobilized onto TiCO₂-coated magnetite suffered from enzyme being removed from the matrix under hydrolysis-use conditions, whereas the PAM enzyme remained attached to the matrix. The physicochemical properties of the immobilizedβ-glucosidase preparations are described. Both immobilizedβ-glucosidase preparations were capable of completely hydrolyzing cellobiose. Recycling of the immobilized enzymes (IME) resulted in reduced rates of hydrolysis with each recycling of the enzyme, although cellobiose was still capable of being completely hydrolyzed. The reduced hydrolysis performance was attributable to physical losses of IME during recovery and, in the case of TAM, enzyme loss from the matrix. Supplementing cellulase digests of steam-explosion pretreatedEucalyptus regnons pulps with immobilizedβ-glucosidase resulted in enhanced hydrolysis. Cellulose-to-glucose yields of 80% of theoretical predictions resulted within 24 h. The magnetically immobilizedβ-glucosidase could easily be recovered from the lignocellulose solids suspension in a stirred batch reactor by applying a magnetic field. The recycled immobilized enzyme continued to convert cellobiose into glucose in 80% yields over a 24-h period. This is the first report of a magnetically immobilizedβ-glucosidase preparation used in the enzymatic saccharification of a lignocellulosic material.     

Cu-Doped ZnO Nanoparticles for Non-Enzymatic Glucose Sensing

Copper-doped zinc oxide nanoparticles (NPs) Cu_xZn_1−xO (x = 0, 0.01, 0.02, 0.03, and 0.04) were synthesized via a sol-gel process and used as an active electrode material to fabricate a non-enzymatic electrochemical sensor for the detection of glucose. Their structure, composition, and chemical properties were characterized using X-ray diffraction (XRD), transmission electron microscopy (TEM), Fourier-transform infrared (FTIR) and Raman spectroscopies, and zeta potential measurements. The electrochemical characterization of the sensors was studied using cyclic voltammetry (CV), electrochemical impedance spectroscopy (EIS), and differential pulse voltammetry (DPV). Cu doping was shown to improve the electrocatalytic activity for the oxidation of glucose, which resulted from the accelerated electron transfer and greatly improved electrochemical conductivity. The experimental conditions for the detection of glucose were optimized: a linear dependence between the glucose concentration and current intensity was established in the range from 1 nM to 100 μM with a limit of detection of 0.7 nM. The proposed sensor exhibited high selectivity for glucose in the presence of various interfering species. The developed sensor was also successfully tested for the detection of glucose in human serum samples.     

Molecular evolution and structural variations in nuclear encoded chloroplast localized heat shock protein 26 (sHSP26) from genetically diverse wheat species

Heat shock proteins are an important class of molecular chaperones known to impart tolerance under high temperature stress. sHSP26, a member of small heat shock protein subfamily is specifically involved in protecting plant’s photosynthetic machinery. The present study aimed at identifying and characterizing sequence and structural variations in sHSP26 from genetically diverse progenitor and non-progenitor species of wheat. In silico analysis identified three paralogous copies of TaHSP26 to reside on short arm of chromosome 4A while one homeologue each was localized on long arm of chromosome 4B and 4D of cultivated bread wheat. Wild DD-genome donor Aegilops tauschii carried an additional sHSP26 gene (AET4Gv20569400) which was absent in the cultivated DD genome of bread wheat. In vitro amplification of this novel gene in wild accessions of Ae. tauschii and synthetic hexaploid wheat but not in cultivated bread wheat validated this finding. Further, significant length polymorphism could be identified in exon1 from diverse sHSP26 sequences. Multiple sequence alignment of procured sequences revealed numerous sSNPs and nsSNPs. D3A, P125 L, Q242 K were designated as homeolog specific- while A49 G as non-progenitor specific amino acid replacements. A 9-bp indel in TmHSP26-1(GA) translated into a deletion of SPM amino acid segment in chloroplast specific conserved consensus region III. High degree of divergence in nucleotide sequence between cultivated and wild species appeared in the form of higher ω values (Ka/Ks >1) indicating positive selection during the course of evolution. Phylogenetic analysis elucidated ancestral relationships between wheat sHSP26 proteins and orthologous proteins across plant kingdom. Overall, data mining approach may be employed as an effective pre-breeding strategy to identify and mobilize novel stress responsive genes and distinct allelic variants from wider germplasm collections of wheat to enhance climate resilience of present day elite wheat cultivars.