Structural identification of novel oligosaccharides produced by Lactobacillus bulgaricus and Lactobacillus plantarum

β-Galactosidases (β-Gal) of lactic acid bacteria produce oligosaccharides from lactose when suitable acceptor carbohydrates are present. This study aimed to elucidate the structure of oligosaccharides formed by galactosylation of N-acetylglucosamine (GlcNAc) and fucose. Crude cellular extract of Lactobacillus bulgaricus and LacLM of Lactobacillus plantarum were used as sources of β-Gal activity. Disaccharides obtained by galactosylation of GlcNAc were identified as Gal-β-(1→4)-GlcNAc or Gal-β-(1→6)-GlcNAc by liquid chromatography-tandem mass spectrometry (LC-MS/MS) and comparison with external standards. Trisaccharides were identified as Gal-β-(1→6)-Gal-β-(1→[4 or 6])-GlcNAc by LC-MS, analysis of the MS/MS spectra of selected in-source fragment ions, and their relative retention times. LC-MS analysis revealed the presence of five galactosylated fucosides, but their linkage type could not be identified, partly due to the lack of reference compounds. β-Gal of lactic acid bacteria may serve as suitable tools for the chemoenzymatic synthesis of therapeutic oligosaccharides.     

Mass production of rubusoside using a novel stevioside-specific β-glucosidase from Aspergillus aculeatus

Rubusoside (R) is a natural sweetener and a solubilizing agent with antiangiogenic and antiallergic properties. However, currently, its production is quite expensive, and therefore, we have investigated nine commercially available glycosidases to optimize an economically viable R-production method. A stevioside (ST)-specific β-glucosidase (SSGase) was selected and purified 7-fold from Aspergillus aculeatus Viscozyme L by a two-step column chromatography procedure. The 79 kDa protein was stable from pH 3.0 to pH 7.0 at 50-60 °C. Hydrolysis of ST by SSGase produced R and steviol monoglucosyl ester as determined by (1)H and (13)C nuclear magnetic resonance (NMR). Importantly, SSGase showed higher activity toward ST than other β-linked glucobioses. The optimal conditions for R production were 280 mM ST and 16.6 μL of SSGase at pH 5.1 and 63 °C. This is the first discussion detailing the production of R by enzymatic hydrolysis of ST and is useful for the food additive and pharmaceutical industries.     

Antiproliferative Activity and Apoptosis Induction of Eucalyptus Citriodora Resin and Its Major Bioactive Compound in Melanoma B16F10 Cells

Antiproliferative activity and apoptosis induction of ethyl acetate of Eucalyptus citriodora resin (EAEER), and its major bioactive compound in melanoma B16F10 cells were investigated. 6-[1-(p-Hydroxy-phenyl)ethyl]-7-O-methyl aromadendrin (HEMA), a flavanol derivative, was isolated from EAEER and identified on the basis of its mass and NMR spectra. The results from MTT assay showed high antiproliferative effects of EAEER and HEMA on B16F10 cells. Moreover, EAEER- and HEMA-induced cell apoptosis was association with the decrease in the mitochondrial transmembrane potentials (Δψ(m)), increase in Bax/Bcl-2 ratio, and activation of caspase-3. Cells treated with EAEER and HEMA generated intracellular reactive oxygen species (ROS) and nitric oxide (NO), indicating that ROS and RNS play important roles in the induction of apoptosis in B16F10 cells. Taken together, EAEER and its major bioactive compound, HEMA, inhibited the proliferation of B16F10 cells via apoptosis and may be a potential antimelanoma agent.     

Metabolic Profiling of Glucosinolates, Anthocyanins, Carotenoids, and Other Secondary Metabolites in Kohlrabi (Brassica oleracea var. gongylodes)

We profiled and quantified glucosinolates, anthocyanins, carotenoids, and other secondary metabolites in the skin and flesh of pale green and purple kohlrabis. Analysis of these distinct kohlrabis revealed the presence of 8 glucosinolates, 12 anthocyanins, 2 carotenoids, and 7 phenylpropanoids. Glucosinolate contents varied among the different parts and types of kohlrabi. Glucoerucin contents were 4-fold higher in the flesh of purple kohlrabi than those in the skin. Among the 12 anthocyanins, cyanidin 3-(feruloyl)(sinapoyl) diglucoside-5-glucoside levels were the highest. Carotenoid levels were much higher in the skins than the flesh of both types of kohlrabi. The levels of most phenylpropanoids were higher in purple kohlrabi than in pale green ones. trans-Cinnamic acid content was 12.7-fold higher in the flesh of purple kohlrabi than that in the pale green ones. Thus, the amounts of glucosinolates, anthocyanins, carotenoids, and phenylpropanoids varied widely, and the variations in these compounds between the two types of kohlrabi were significant.     

Development of an Environmentally Friendly Adsorbent for the Removal of Toxic Heavy Metals from Aqueous Solution

An effective adsorbent for the removal of heavy metals was manufactured by immobilization of jujube powder. The adsorptions of Cd, Zn and Cu from aqueous solutions by jujube complex beads (Type 1 and Type 2) were studied in a batch adsorption system. The adsorption data were fitted well with the Langmuir isotherm models. The adsorption capacities (β) for Cd, Zn and Cu were 4.23, 2.93 and 3.64 mg/g in Type 1 and 1.24, 0.70 and 1.35 mg/g in Type 2 beads. The removal efficiencies of the Type 2 beads, with a larger unit surface area, were lower than those of the Type 1 due to part of the casein or cyclic AMP being destroyed during the drying process of the Type 1. These values for Type 1 beads were higher than those of all other adsorbents for each heavy metal. A comparison of the kinetic models on the overall adsorption rate showed that the adsorption system was best described by pseudo-first-order kinetics. The removal efficiencies of Cd, Zn and Cu exhibited similar tendencies to those observed in the equilibrium tests. This indicates that the jujube complex beads developed in this study can be used as promising adsorbents for the removal of heavy metals from wastewater.     

Simultaneous Removal of Phosphate and Nitrate in Wastewater Using High-Capacity Anion-Exchange Resin

The removal of nutrients in wastewater is a critical issue in water treatment because released nutrients can cause serious adverse effects in water systems or the aquatic environment. In this study, a high-performing polymeric anion-exchange resin was developed for the removal of nutrients from wastewater. The resins were prepared by chloromethylation followed by amination under preferred conditions. The resins were investigated for the removal of nutrients such as phosphate, nitrate, and fluoride from water. The density of functional groups on the synthesized resin was approximately 33?% higher than on commercialized resins, and the specific surface area of synthesized resin was increased by approximately tenfold compared with commercialized resins. The adsorption capacity of synthesized resins (AMP16-FeCl₃) for anions was 285.8?mg/g, which was approximately three times higher than the capacity of commercialized resin (AMP16-OH). A study of the effects of the types of counterions and functional groups found that resins having hydroxide ion as a counterion showed higher capacity and selectivity for phosphate ion and that dimethylethanolamine as a resin functional group of resin was more efficient than trimethylamine.     

Different response of silicate fertilizer having electron acceptors on methane emission in rice paddy soil under green manuring

Cultivation of green manure plants during the fallow season in rice paddy soil has been strongly recommended to improve soil properties. However, green manuring may impact greenhouse gas emission, methane (hereafter, CH₄) in particular, under the flooded rice cultivation and thus, application of chemical amendments being electron acceptors can be an effective mitigation strategy to reduce CH₄ emissions in irrigated rice (Oryza sativa L.) field amended with green manure. To investigate the effect of iron (Fe) slag silicate fertilizer (hereafter, silicate fertilizer), which was effective in reducing CH₄ emission and increasing rice productivity, in green manure-amended paddy soil, the aboveground biomass of Chinese milk vetch (hereafter, vetch) was added at rates of 0, 10, 20, and 40 Mg (fresh weight) ha⁻¹ before the application of silicate fertilizer, which was added at rates of 0 and 2.3 Mg ha⁻¹. Silicate fertilization reduced the seasonal CH₄ flux by ca. 14.5% and increased rice yield by ca. 15.7% in the control (no vetch application) treatment. However, CH₄ production was increased by silicate fertilization in vetch-treated soil particularly at the initial rice growing stage, which was probably due to the enhanced decomposition of added organic matters by the silicate liming effect. In conclusion, silicate fertilization is not effective in reducing CH₄ production in green manure-amended rice paddy soils and its use should be properly controlled.