An extracellular lipase from Aureobasidium pullulans was obtained and purified with a specific activity of 17.7 U/mg of protein using ultrafiltration and a DEAE-Sepharose Fast Flow column. Characterization of the lipase indicated that it is a novel finding from the species A. pullulans. The molecular weight of the lipase was 39.5 kDa, determined by sodium dodecyl sulfonate-polyacrylamide gel electrophoresis (SDS-PAGE). The enzyme exhibited its optimum activity at 40 °C and pH of 7. It also showed a remarkable stability in some organic solutions (30%, v/v) including n-propanol, isopropanol, dimethyl sulfoxide (DMSO), and hexane. The catalytic activity of the lipase was enhanced by Ca²⁺ and was slightly inhibited by Mn²⁺ and Zn²⁺ at a concentration of 10 mmol/L. The lipase was activated by the anionic surfactant SDS and the non-ionic surfactants Tween 20, Tween 80, and Triton X-100, but it was drastically inhibited by the cationic surfactant cetyl trimethyl ammonium bromide (CTAB). Furthermore, the lipase was able to hydrolyze a wide variety of edible oils, such as peanut oil, corn oil, sunflower seed oil, sesame oil, and olive oil. Our study indicated that the lipase we obtained is a potential biocatalyst for industrial use.
Ascomycota/enzymology* ; Calcium ; Catalysis ; Corn Oil/metabolism* ; Detergents/chemistry* ; Enzyme Stability ; Fungal Proteins/chemistry* ; Glucans/chemistry* ; Hexanes/chemistry* ; Hydrogen-Ion Concentration ; Hydrolysis ; Industrial Microbiology ; Lipase/chemistry* ; Manganese/chemistry* ; Olive Oil/metabolism* ; Peanut Oil/metabolism* ; Sesame Oil/metabolism* ; Substrate Specificity ; Sunflower Oil/metabolism* ; Surface-Active Agents ; Temperature ; Zinc/chemistry*

The transplants of one-year-old Glycyrrhiza uralensis Fisch. were subjected to five concentrations of MnSO4-H2O (0, 1.81, 18.1, 36.2 and 54.3 mg·L(-1)) culturing in vermiculite. qRT-PCR and HPLC were respectively used to measure the relative expression of SQS1 gene and the content of glycyrrhizic acid of G. uralensis in different concentrations of MnSO4·H2O. This is to explore discuss the effects of the expression of SQS1 gene and the accumulation of glycyrrhizic acid by Mn treatment. The results showed both the expression of SQS1 gene and the content of glycyrrhzic acid of G. uralensis tended to rise after the fall of the first with the increase of concentration of Mn treatment. And they were of very significant positive correlation (P<0.01, r=0.737). Relative expression of SQS1 gene reached the highest 7.90 under 18.1 mg·L(-1) MnSO4·H2O treatment. It was very significantly different between 18.1 mg·L(-1) concentration of MnSO4·H2O treatment and CK (0 mg·L(-1)), 1.81, 36.2 and 54.3 mg·L(-1) (P<0.01), and 1.75, 1.37, 1.37, 2.33 times respectively. The content of glycyrrhizic acid reached the highest under 1.81 and 18.1 mg·L(-1) MnSO4·H2O treatment, and there were not significant difference (P>0.05). It was very significantly different between them and other concentrations of MnSO4·H2O treatment (P<0.01). This study suggests the appropriate concentration of Mn treatment could certain promote the expression of SQS1 gene and the accumulation of glycyrrhizic acid of G. uralensis.
Chromatography, High Pressure Liquid ; Genes, Plant ; Glycyrrhiza uralensis ; chemistry ; genetics ; Glycyrrhizic Acid ; analysis ; Manganese

OBJECTIVETo investigate the effects of polygala on leaning and memory and the expression of Microtubule associated protein on manganese poisoned mice.

METHODS60 female Kunming mice were randomly and equally divided into 5 group. They are normal control group (CG), manganese poisoned group (MG), manganese poisoned with polygala high dose group (MHG), manganese poisoned with polygala middle dose group (MMG), manganese poisoned with polygala low dose group (MLG). The model of manganese poisoned mice was prepared of the way of intraperitoneal injection of manganese chloride (MnCl2 15 mg/kg), the spatial learning and memory ability was tested by Morris water maze, the Doublecortin (DCX) was tested by the way of immunofluorescent staining in the SVZ and SGZ.

RESULTIn the navigation test, compared with MG, the escape latency of MHG, MMG and MLG were significantly decreased (P < 0.05), in space exploration experiments, MHG, MMG, MLG compared with MG, the number increased significantly across platforms (P < 0.05). compared with MG, the DCX expression of MHG, MMG and MLG were significantly increased (P < 0.05).

CONCLUTIONThe leaning and memory ability of manganese poisoned mice can be improved by the polygala, and the mechanism may be related to promote the expression of DCX and neurogenesis in the brain.

Animals ; Female ; Manganese Poisoning ; drug therapy ; Maze Learning ; drug effects ; Memory ; drug effects ; Mice ; Microtubule-Associated Proteins ; drug effects ; Neurogenesis ; drug effects ; Neuropeptides ; drug effects ; Plant Extracts ; pharmacology ; Polygala ; chemistry

To synthesize and characterize a novel metal complex of Mn (II) with emodin, and evaluate its anti-cancer activity. The elemental analyses, IR, UV-vis, atomic absorption spectroscopy, TG-DSC, (1)H NMR, and (13)C NMR data were used to characterize the structure of the complex. The cytotoxicity of the complex against the human cancer cell lines HepG2, HeLa, MCF-7, B16, and MDA-MB-231 was tested by the MTT assay and flow cytometry. Emodin was coordinated with Mn(II) through the 9-C=O and 1-OH, and the general formula of the complex was Mn(II) (emodin)2·2H2O. In studies of the cytotoxicity, the complex exhibited significant activity, and the IC50 values of the complex against five cancer cell lines improved approximately three-fold compared with those of emodin. The complex could induce cell morphological changes, decrease the percentage of viability, and induce G0/G1 phase arrest and apoptosis in cancer cells. The coordination of emodin with Mn(II) can improve its anticancer activity, and the complex Mn(II) (emodin)2·2H2O could be studied further as a promising anticancer drug.
Antineoplastic Agents ; chemical synthesis ; pharmacology ; therapeutic use ; Antineoplastic Agents, Phytogenic ; pharmacology ; therapeutic use ; Emodin ; pharmacology ; therapeutic use ; HeLa Cells ; Hep G2 Cells ; Humans ; MCF-7 Cells ; Manganese ; pharmacology ; therapeutic use ; Melanoma, Experimental ; drug therapy ; Molecular Structure ; Neoplasms ; drug therapy ; Phytotherapy ; Plant Extracts ; pharmacology ; therapeutic use ; Polygonaceae ; chemistry

We measured selenium, zinc, copper and manganese concentrations in the human milk of Korean mothers who gave birth to preterm infants, and compared these measurements with the recommended daily intakes. The samples of human milk were collected postpartum at week-1, -2, -4, -6, -8, and -12, from 67 mothers who gave birth to preterm infants (< 34 weeks, or birth weight < 1.8 kg). All samples were analyzed using atomic absorption spectrophotometry. The concentrations of selenium were 11.8 +/- 0.5, 11.4 +/- 0.8, 12.7 +/- 0.9, 11.4 +/- 0.8, 10.8 +/- 0.9, and 10.5 +/- 1.3 microg/L, zinc were 7.8 +/- 0.5, 9.1 +/- 0.8, 7.2 +/- 0.9, 8.0 +/- 0.8, 7.4 +/- 0.9, and 6.6 +/- 1.2 mg/L, copper were 506 +/- 23.6, 489 +/- 29.4, 384 +/- 33.6, 356 +/- 32.9, 303 +/- 35.0, and 301 +/- 48.0 microg/L and manganese were 133 +/- 4.0, 127 +/- 6.0, 125 +/- 6.0, 123 +/- 6.0, 127 +/- 6.0, and 108 +/- 9.0 microg/L at week-1, -2, -4, -6, -8, and -12, respectively. The concentrations of selenium and zinc meet the daily requirements but that of copper is low and of manganese exceeds daily requirements recommended by the American Academy of Pediatrics, Committee on Nutrition.
Adult ; Copper/analysis ; Female ; Humans ; Infant, Newborn ; Infant, Premature ; Longitudinal Studies ; Manganese/analysis ; Milk, Human/*chemistry ; Postpartum Period ; Republic of Korea ; Selenium/analysis ; *Spectrophotometry, Atomic ; Trace Elements/*analysis ; Zinc/analysis

OBJECTIVE: To establish a new method to estimate injury-tool by analyzing the components of left metal particles from hammers impact on pig skin and filter paper using scanning electron microscope and energy dispersive X-ray spectrometer (SEM-EDX). METHODS: The pig skin and filter paper were stricken by two steel hammers. The left metal particles were examined by SEM-EDX and the results were statistically analyzed by SPSS 13.0. RESULTS: The characteristics of left particles showed stable by several impacts using one steel hammer. The left particles showed no statistical difference for impact on pig skin and filter paper. The left particles displayed a statistical difference using two hammer with different components. CONCLUSION SEM-EDX can be used to detect the left metal particles from the steel hammer and can provide a method for estimating injury-tool.
Animals ; Forensic Medicine/methods* ; Iron/analysis* ; Manganese/analysis* ; Metals/chemistry* ; Microscopy, Electron, Scanning/methods* ; Models, Animal ; Skin/injuries* ; Spectrometry, X-Ray Emission/methods* ; Swine ; Wounds, Nonpenetrating/pathology*

D-psicose 3-epimerase (DPEase) is demonstrated to be useful in the bioproduction of D-psicose, a rare hexose sugar, from D-fructose, found plenty in nature. Clostridium cellulolyticum H10 has recently been identified as a DPEase that can epimerize D-fructose to yield D-psicose with a much higher conversion rate when compared with the conventionally used DTEase. In this study, the crystal structure of the C. cellulolyticum DPEase was determined. The enzyme assembles into a tetramer and each subunit shows a (β/α)(8) TIM barrel fold with a Mn(2+) metal ion in the active site. Additional crystal structures of the enzyme in complex with substrates/products (D-psicose, D-fructose, D-tagatose and D-sorbose) were also determined. From the complex structures of C. cellulolyticum DPEase with D-psicose and D-fructose, the enzyme has much more interactions with D-psicose than D-fructose by forming more hydrogen bonds between the substrate and the active site residues. Accordingly, based on these ketohexose-bound complex structures, a C3-O3 proton-exchange mechanism for the conversion between D-psicose and D-fructose is proposed here. These results provide a clear idea for the deprotonation/protonation roles of E150 and E244 in catalysis.
Binding Sites ; Biocatalysis ; Catalytic Domain ; Clostridium cellulolyticum ; enzymology ; Hexoses ; chemistry ; Manganese ; chemistry ; Protein Structure, Quaternary ; Racemases and Epimerases ; chemistry ; metabolism ; Substrate Specificity

Little is known about hair mineral status in fibromyalgia patients. This study evaluated the characteristics of hair minerals in female patients with fibromyalgia compared with a healthy reference group. Forty-four female patients diagnosed with fibromyalgia according to the American College of Rheumatology criteria were enrolled as the case group. Age- and body mass index-matched data were obtained from 122 control subjects enrolled during visit for a regular health check-up. Hair minerals were analyzed and compared between the two groups. The mean age was 43.7 yr. General characteristics were not different between the two groups. Fibromyalgia patients showed a significantly lower level of calcium (775 microg/g vs 1,093 microg/g), magnesium (52 microg/g vs 72 microg/g), iron (5.9 microg/g vs 7.1 microg/g), copper (28.3 microg/g vs 40.2 microg/g) and manganese (140 ng/g vs 190 ng/g). Calcium, magnesium, iron, and manganese were loaded in the same factor using factor analysis; the mean of this factor was significantly lower in fibromyalgia group in multivariate analysis with adjustment for potential confounders. In conclusion, the concentrations of calcium, magnesium, iron, and manganese in the hair of female patients with fibromyalgia are lower than of controls, even after adjustment of potential confounders.
Adult ; Body Height ; Body Mass Index ; Calcium/analysis ; Female ; Fibromyalgia/*metabolism ; Hair/*chemistry ; Humans ; Iron/analysis ; Magnesium/analysis ; Manganese/analysis ; Metals/*analysis ; Middle Aged

The effects of reaction temperature, ethanol concentration and weight hourly space velocity (WHSV) on the ethylene production from ethanol dehydration using zinc, manganese and cobalt modified HZSM-5 catalyst were investigated by response surface methodology (RSM). The results showed that the most significant effect among factors was reaction temperature and the factors had interaction. The optimum conditions were found as 34.4% ethanol concentration, 261.3 0 degrees C of reaction temperature and 1.18 h(-1) of WHSV, under these conditions the yield of ethylene achieved 98.69%.
Catalysis ; Cobalt ; chemistry ; Dehydration ; Ethanol ; chemistry ; Ethylenes ; chemistry ; metabolism ; Manganese ; chemistry ; Zeolites ; chemistry ; Zinc ; chemistry

Human NUDT16 (hNUDT16) is a decapping enzyme initially identified as the human homolog to the Xenopus laevis X29. As a metalloenzyme, hNUDT16 relies on divalent cations for its cap-hydrolysis activity to remove m⁷GDP and m²²⁷GDP from RNAs. Metal also determines substrate specificity of the enzyme. So far, only U8 small nucleolar RNA (snoRNA) has been identified as the substrate of hNUDT16 in the presence of Mg²(+). Here we demonstrate that besides U8, hNUDT16 can also actively cleave the m⁷GDP cap from mRNAs in the presence of Mg²(+) or Mn²(+). We further show that hNUDT16 does not preferentially recognize U8 or mRNA substrates by our cross-inhibition and quantitative decapping assays. In addition, our mutagenesis analysis identifies several key residues involved in hydrolysis and confirms the key role of the REXXEE motif in catalysis. Finally an investigation into the subcellular localization of hNUDT16 revealed its abundance in both cytoplasm and nucleus. These findings extend the substrate spectrum of hNUDT16 beyond snoRNAs to also include mRNA, demonstrating the pleiotropic decapping activity of hNUDT16.
Amino Acid Motifs ; Biocatalysis ; Cell Nucleus ; enzymology ; Consensus Sequence ; Cytoplasm ; enzymology ; metabolism ; Guanosine Diphosphate ; metabolism ; Histidine ; metabolism ; Humans ; Hydrolysis ; Luciferases ; genetics ; Magnesium ; metabolism ; Manganese ; metabolism ; Mutagenesis ; Mutation ; Pyrophosphatases ; antagonists & inhibitors ; chemistry ; genetics ; metabolism ; RNA Caps ; chemistry ; metabolism ; pharmacology ; RNA, Small Nucleolar ; chemistry ; metabolism ; pharmacology