Cloning and analysis of a Y-tocopherol methyltransf erase gene from Brassica oleracea and the function of its recombinant protein

The full-length cDNA (BoTMT) of γ-TMT is obtained from Brassica oleracea by 3'- and 5'-RACE methods. The 1265 bp cDNA contains an open reading frame of 1044 bp, which encodes a protein of 347 amino acids with a predicted chloroplast transit peptide and two S-adenosylmethionine (SAM)-binding domains. Sequence analysis shows that the deduced protein shares 41. 8% -86. 5% similarity to known γ-TMTs. Semi-quantitative RT-PCR reveals that BoTMT is expressed preferentially in flowers and leaves of B. oleracea . The recombinant γ-TMT protein is obtained by cloning its encoding region into the prokaryotic expression vector pET30a. The protein expressed in E. coli accounts for 22 % of total bacterial protein. The enzyme activity assay indicates that the recombinant protein has relatively high activity to convert γ-tocopherol to α-tocopherol in vitro.     

Cloning and analysis of a γ-tocopherol methyltransferase gene from Brassica oleracea and the function of its recombinant protein

The full-length cDNA (BoTMT) of γ-TMT is obtained from Brassica oleracea by 3′- and 5′-RACE methods. The 1265 bp cDNA contains an open reading frame of 1044 bp, which encodes a protein of 347 amino acids with a predicted chloroplast transit peptide and two S-adenosylmethionine (SAM)-binding domains. Sequence analysis shows that the deduced protein shares 41.8%～86.5% similarity to known γ-TMTs. Semi-quantitative RT-PCR reveals that BoTMT is expressed preferentially in flowers and leaves of B. oleracea. The recombinant γ-TMT protein is obtained by cloning its encoding region into the prokaryotic expression vector pET30a. The protein expressed in E. coli accounts for 22% of total bacterial protein. The enzyme activity assay indicates that the recombinant protein has relatively high activity to convert γ-tocopherol to α-tocopherol in vitro.     

Construction and characterization of partially ntrC-deleted mutants in Alcaligenes faecalis

To study the effect of ntrC gene product on the expression and regulation of other important nitrogen-fixing genes in Alcaligenes faecalis, partially ntrC-deleted mutants of A. faecalis have been generated. To start with, the ntrC gene of A. faecalis was cloned into a suicide plasmid pSUP202 to create a recombinant plasmid pSUM1. The ntrC gene in pSUM1 was then replaced by a lacZ-Kmr fragment resulted in the generation of a plasmid pSUM2. The lacZ fragment in pSUM2 was further removed and a plasmid pSUM3 produced. As a second step, the plasmid pSUM2 or pSUM3 was introduced into the wild type of A. faecalis A1501 by conjugation and two partially ntrC-deleted mutants A15CM1 (ntrC∷lacZ) and A15CM2 (ntrC-) were obtained. To understand the regulatory effect of the NtrC on the expression of nifH and nifA, a nifH-lacZ gene or a nifA-lacZ gene was introduced into the ntrC- mutant by conjugation. The results indicated that: (ⅰ) although the ntrC- mutant was nif + , its nitrogen fixation activity was only 20% that of the wild type; (ⅱ) the ntrC- mutant failed to grow on the medium containing nitrate as a sole nitrogen source; (ⅲ) the regulation of ntrC gene expression did not require its own product; (ⅳ) the expression of nifH in A . faecalis was positively regulated by the ntrC. Deletion of the ntrC resulted in the reduction of nifH expression or even totally inactivated nitrogen fixation; (ⅴ) there was no obvious influence on the expression of nifA in A. faecalis if the ntrC gene was deleted.     

2,3-Dihydroxybiphenyl dioxygenase gene was first discovered in Arthrobacter sp. strain PJ3

Bacterium strain PJ3,isolated from wastewater and identified as Arthrobacter sp. bacterium based on its 16S rDNA gene,could use carbazole as the sole carbon,nitrogen and energy source. The genomic library of strain PJ3 was constructed and a positive clone JM109(pUCW402) was screened out for the expression of dioxygenase by the ability to form yellow ring-fission product. A 2,3-dihydroxybiphenyl dioxygenase(23DHBD) gene of 933 bp was found in the 3360 bp exogenous fragment of pUCW402 by GenSCAN software and BLAST analysis. The phylogenetic analysis showed that 23DHBD from strain PJ3 formed a deep branch separate from a cluster containing most known 23DHBD in GenBank. Southern hybridization confirmed for the first time that the 23DHBD gene was from the genomic DNA of Arthrobacter sp. PJ3. In order to test the gene function,recombinant bacterium BL21(pETW-8) was constructed to express 23DHBD. The expression level in BL21(pETW-8) was highest compared with the recombinant bacteria JM109(pUCW402) and strain PJ3. We observed that 23DHBD was not absolute specific. The enzyme activity was higher with 2,3-dihydroxybiphenyl as a substrate than with catechol. The substrate specificity assay suggested that 23DHBD was essential for cleavage of bi-cyclic aromatic compounds during the course of aromatic compound biodegradation in Arthrobacter sp. strain PJ3.     

Corrosion behavior of high-strength spring steel for high-speed railway

The corrosion resistance and evolution of corrosion products in medium-carbon high-strength spring steels were investigated in a neutral salt spray (5wt% NaCl solution). A formation model of γ-FeOOH and a transformation model describing the conversion of γ-FeOOH to α-FeOOH were constructed. The results indicated that, at the initial corrosion stage, the corrosion resistance was gradually improved with the addition of Cr; however, with the addition of alloying element V, the corrosion resistance decreased. These results were attributed mainly to the initial corrosion stage being closely related to the matrix microstructure parameters such as grain-boundary character and dislocation density. After the rust layer was formed at a later corrosion stage, the corrosion resistance was reinforced with the addition of Cr and V because Cr strongly influenced the composition, structure, and morphology of the corrosion products. The results presented herein show that Cr was conducive to the transformation of γ-FeOOH into α-FeOOH. Moreover, V and Cr exhibited obvious synergy and were enriched in the inner layer of the corrosion products.     

Influence of the Mode of Wavelength on the Infrared Reflectance and Transmittance in the Fabric

The study and the practical utilization of the infrared inthe textile field is analysed simply.The test method ofthe infrared transmittance(α_T)and the reflectance(α_R)in the fabric is introduced.The test results of aset of polyester/cotton plain fabrlcs show that the infra-red reflecting regulation can be expressed by quasi-co-sine function.The parameters affecting the α_T and α_Rare analysed, α_T decreases according to negative expo-nent function as the weight per square meter of the fab-ric increases,and α_R increases.Both α_T and α_R will in-crease when the mode of wavelength(λ_m) of the infra-red decreases.It can be deduced that the infrared ab-sorptance of the fabric will increase as λ_m increases inthe studied range.     

Characterization of γ-Fe2O3 nanoparticles prepared by transfor- mation of α-FeOOH

γ-Fe2O3 nanoparticles were successfully synthesized by a chemically induced transformation of α-FeOOH.In this method,the precursor(α-FeOOH)was prepared by chemical precipitation,and then treated with a mixed FeCl2/NaOH solution to produce the nanoparticles.X-ray diffraction indicated that when the precursor was treated with FeCl2(0.22 mol/L)and NaOH(0.19 mol/L),pure γ-Fe2O3 nanoparticles were obtained.However,when the concentration of FeCl2 was<0.22 mol/L or the concentration of NaOH was<0.19 mol/L,α-FeOOH and γ-Fe2O3 phases co-existed in the nanoparticles.Transmission electron microscopy observations showed that in the samples with co-existing phases,the nanoparticles did not have identical morphologies.The pure γ-Fe2O3 nanoparticles were polygonal rather than spherical.The volume ratio of α-FeOOH and γ-Fe2O3 was estimated for the two-phase samples from magnetization data obtained from a vibrating sample magnetometer.This chemically induced transformation is novel,and could provide an effective route for the synthesis of other metal oxide nanocrystallites.     

Rudimentary study on humanization of porcine red blood cells:Enzymatic removal of galactose-α1,3-galactose antigen from porcine red blood cell

The serological and biochemical characterization of porcine red blood cells (pRBCs) are similar to human red blood cells,Porcine erythrocytes are considered as an alternative source for human blood transfusion.But there exist galactose-α1,3-galactose antigens (Galα1,3Galβ1,4galNAc-R,abbreviated αGal antigen ) on pRBCs,which can induce anti-αGai antibodies in human serum ,The αGal epitopes are the major antigen responsible for hyperacute rejection in exnotransfusion .In this study ,recombined soybean α-galactosidase (rSα-GalE) was used to remove the αGal antigens from pPRCs for humanization .The results showed that αGal antigen was eleared by rSα-GalE and the structure and function of rSα-GalE treated pRBC were normal.     

Evidence from 18O feeding studies for hydroxyl group donor in the reaction catalyzed by cytidylate hydroxymethylase MilA

5-Hydroxymethylcytosine (5hmC) was present in T-even phage and mammalian DNA. 5hmC in phage is formed by hydroxymethylation of the cytosine base in deoxycytidylate (dCMP) by deoxycytidylate hydroxymethylase (CH), which uses the solvent water as the hydroxyl group donor. By contrast, 5hmC is formed in mammal zygotes by the oxidation of 5-methylcytosine (5mC). 5hmC was also present in a nucleoside antibiotic mildiomycin and its formation is governed by a cytidylate hydroxymethylase MilA. However, the catalytic mechanism remains unknown. In the present study, we purified His-tagged MilA and fed its in vitro reaction with H218O. The LC-MS analysis of the product revealed that 18O was incorporated into the hydroxymethylated CMP (HmCMP), and the secondary MS result of 18O-labeled HmCMP indicated that 18O was incorporated into the cytosine of HmCMP. The results demonstrate that MilA uses solvent water as the hydroxyl group donor like CH. Moreover, Thr102 of MilA was predicted as potential critical amino acid anchoring one molecule of water for hydroxylation. Finally, organizational context comparison in microbial genomes reveals that six homologous ORFs originally annotated as putative thymidylate synthase (TS) are more likely to be CMP hydroxymethylase.     

Epoxy/α-alumina nanocomposite with high electrical insulation performance

An experimental study was conducted to improve the electrical insulation of epoxy resin. The effects of boehmite, γ-alumina and α-alumina nanoparticles on the volume resistivity, dielectric strength and glass transition temperature of epoxy nanocomposites were investigated. The results showed that α-alumina nanoparticles displayed obvious advantages in enhancing electrical insulation performance of epoxy nanocomposites, compared to boehmite and γ-alumina nanoparticles. The direct current volume resistivity and breakdown strength of epoxy nanocomposite with 2.0 wt% α-alumina nanoparticles was improved to 2.2 × 10~(18)Ω cm and76.1 kV mm~(-1) respectively. And these improved values of electrical insulation properties are much higher than these of epoxy nanocomposites reported in previous studies. The main reason of these improvements may be that the epoxy/α-alumina interaction zone was enhanced by crosslink.     

Enhanced Biological Phosphorus Removal in SBR Using Glucose as a Single Organic Substrate

Enhanced biological phosphorus removal （EBPR） was investigated in an anaerobic/aerobic sequencing batch reactor （SBR） supplied with glucose as a single organic substrate. The results illustrated that EBPR process could also occur successfully with glucose other than short chain fatty acids （SCFAs）. High phosphorus release and polyhydroxyalkanoate （PHA） accumulation in the anaerobic phase was found vital for the removal of phosphorus during the aerobic phase. The measurement of intracellular reserves revealed that glycogen had a higher chance to replace the energy role of poly-P under anaerobic conditions. Moreover, glycogen was also utilized as the carbon source for PHA synthesis, as well as a reducing power as reported earlier. The accumulated PHA in this system was mainly in the form of poly-hydroxyvalerate （PHV） instead of poly-hydroxybutyrate （PHB）, and was inferred to be caused by the excess reducing power contained in glucose. Lactate as a fermentation product was also found released into the bulk solution. Applying fundamental biochemistry knowledge to the experimental results, a conceptual biochemical model was developed to explain the metabolism of the glucoseinduced EBPR.     

Preparation of a cationic azobenzene dye-montmorillonite intercalation compound and its photochemical behavior

Montmorillonite/cationic azobenzene dye (GTL) intercalation compounds were prepared by the conventional ion exchange method. As compared with that of pure GTL, the thermal stability of the intercalated GTL was greatly enhanced, and the absorption band corresponding to azobenzene group in intercalated GTL shifted towards a longer wavelength by 55 nm, which could be ascribed to the strong conjugation of GTL supramolecular order structure (J cluster) confined in a nanoscale space of montmorillonite interlayer gallery. The microstructures of the resulting intercalation compounds could be successfully controlled by varying the amount of dye loaded as evidenced by the basal spacing of the intercalation compounds. The intercalated azo dye in the montmorillonite interlayer space exhibited reversible trans-to-cis photoisomerization and thermal cis-to-trans reaction. FTIR proved the successful intercalation of GTL into the silicate layer.     

Glycyrrhetinic acid-modified nanoparticles for drug delivery: Preparation and characterization

In this work, glycyrrhetinic acid-modified chitosan （mGA-suc-CTS） used as liver targeted carrier for drug delivery, was prepared via hemisuccinate as a bridged group. The structure of the product was confirmed by IR and NMR methods and the degree of substitution （DS） of glycyrrhetinic acid groups was estimated via elemental analysis. Nanoparticles were formed by ionic gelation methold. The drug-loading and release behavior of the nanoparticles were investigated using BSA as the model drug. The results indicated that the carrier with a highest DS of 5.19% could be got and the DS was controlled by changing reaction temperature or feed ratio. BSA could be entrapped into the nanoparticles with the drug-loading ratio of 26.3% and the encapsulation efficiency of 81.5%. A sustained release over an 11-day period was observed in pH 7.4 in vitro.     

Effect of niobium alloying level on the oxidation behavior of titanium aluminides at 850°C

This work addresses the alloying of titanium aluminides used in aircraft engine applications and automobiles. The oxidation resistance behavior of two titanium aluminides of α₂ + γ(Ti₃Al + TiAl) and orthorhombic Ti₂NbAl, recognized as candidates for high-temperature applications, was investigated by exposure of the alloys for 100 h in air. Thus, oxidation resistance was expressed as the mass gain rate, whereas surface aspects were analyzed using scanning electron microscopy in conjunction with energy-dispersive X-ray spectroscopy, and the type of oxidation products was analyzed by X-ray diffraction and Raman spectroscopy. The orthorhombic Ti₂NbAl alloy was embrittled, and pores and microcracks were formed as a result of oxygen diffusion through the external oxide layer formed during thermal oxidation for 100 h.     

Microstructure evolution and mechanical properties of a large-sized ingot of Mg–9Gd–3Y–1.5Zn–0.5Zr(wt%) alloy after a lower-temperature homogenization treatment

In this paper,a large-sized ingot of Mg–9Gd–3Y–1.5Zn–0.5Zr(wt%) alloy with a diameter of 600 mm was successfully prepared by the semi-continuous casting method.The alloy was subsequently annealed at a relatively low temperature of 430°C for 12 h as a homogenization treatment.The microstructure and room-temperature mechanical properties of the alloy were investigated systematically.The results show that the as-cast alloy contained a mass of discontinuous lamellar-shaped 18 R long-period stacking ordered(LPSO) phases with a composition of Mg10 Zn Y and an α-Mg matrix,along with net-shaped Mg5(Y,Gd) eutectic compounds at the grain boundaries.Most of the eutectic compounds dissolved after the homogenization treatment.Moreover,the amount and dimensions of the lamellar-shaped LPSO phase obviously increased after the homogenization treatment.The structure of the phase transformed into 14H-type LPSO with composition Mg12Zn(Y,Gd).The mechanical properties of the heat-treated large-sized alloy ingot are uniform.The ultimate tensile strength(UTS) and tensile yield strength(TYS) of the alloy reached 207.2 MPa and 134.8 MPa,respectively,and the elongation was 3.4%.The high performances of the large-sized alloy ingot after the homogenization treatment is attributed to the strengthening of the α-Mg solid solution and to the plentiful LPSO phase distributed over the α-Mg matrix.     

Microstructure evolution and mechanical properties of a large-sized ingot of Mg-9Gd-3Y-1.5Zn-0.5Zr (wt%) alloy after a lower-temperature homogenization treatment

In this paper, a large-sized ingot of Mg-9Gd-3Y-1.5Zn-0.5Zr (wt%) alloy with a diameter of 600 mm was successfully prepared by the semi-continuous casting method. The alloy was subsequently annealed at a relatively low temperature of 430℃ for 12 h as a homogenization treatment. The microstructure and room-temperature mechanical properties of the alloy were investigated systematically. The results show that the as-cast alloy contained a mass of discontinuous lamellar-shaped 18R long-period stacking ordered (LPSO) phases with a composition of Mg₁₀ZnY and an α-Mg matrix, along with net-shaped Mg₅(Y,Gd) eutectic compounds at the grain boundaries. Most of the eutectic compounds dissolved after the homogenization treatment. Moreover, the amount and dimensions of the lamellar-shaped LPSO phase obviously increased after the homogenization treatment. The structure of the phase transformed into 14H-type LPSO with composition Mg₁₂Zn(Y,Gd). The mechanical properties of the heat-treated large-sized alloy ingot are uniform. The ultimate tensile strength (UTS) and tensile yield strength (TYS) of the alloy reached 207.2 MPa and 134.8 MPa, respectively, and the elongation was 3.4%. The high performances of the large-sized alloy ingot after the homogenization treatment is attributed to the strengthening of the α-Mg solid solution and to the plentiful LPSO phase distributed over the α-Mg matrix.     

B→O blood conversion

To maintain a constant supply of “universal blood type”,or group O red blood cells,has benefit in specialized transfusion condition.In this study, α-galactosidase cDNA was cloned from Catimor coffee bean grown in Hainan island,China,by the RT-PCR method.We have constructed a vector for α-galactosidase cDNA expression and transferred α-galactosidase cDNA into Pichia pastoris GS115 cells by electroporation.The recombinant α-galactosidase(r-α-GalE)was purified by cation ion exchange chromatography.After studying the biochemical characters of r-α-GalE we have succeeded in converting human erythrocytes from group B to group O.The animal experiment showed that transfusion of enzymetically converted group O red blood cells(ECORBC)was perfectly safe.     

Interactively interfacial reaction of iron-reducing bacterium and goethite for reductive dechlorination of chlorinated organic compounds

The interactively interfacial reactions between the iron-reducing bacterium （Shewanella decolorationis, S12） and iron oxide （α-FeOOH） were investigated to determine reductive dechlorination transformation of chlorinated organic compounds （chloroform and pentachlorophenol）. The results showed that the interactive system of S12＋ α-FeOOH exhibited relatively high dechlorination rate. By comparison, the S12 biotic system alone had no obvious dechlorination, and the α-FeOOH abiotic system showed low dechlorination rate. The enhanced dechlorination of chloroform and pentachlorophenol in the interactive system of S12＋α-FeOOH was derived from the promoted generation of adsorbed Fe（Ⅱ） by S12. A decrease in redox potential of the Fe （Ⅲ）/Fe （Ⅱ） couple in the interactive reaction system was determined by cyclic voltammetry. Our results will give new insight into interactively interfacial reaction between iron-reducing bacterium and iron oxides for degradation of chlorinated organic compounds under anaerobic condition.     

High temperature creep mechanisms of a single crystal superalloy: A phasefield simulation and microstructure characterization

The high temperature creep behavior of a single crystal Ni-based superalloy was studied by combined experimental and numerical methods. The creep test results showed that the creep curves exhibited a three-stage feature. The qualitative explanations for each stage of the creep curves were carried out based on the microstructure characterizations of γ/γ′ phases and dislocations. An elastoplasticity incorporated phase-field model was developed to provide quantitative understanding on directional coarsening(rafting) of γ′ phase. The simulation results showed that the directionality of γ′ coarsening was induced by both dislocation activity in γchannels and elastic inhomogeneity between γ and γ' phases, therein the dislocation activity played a major role.This findings provide new insights into the design of novel single crystal superalloys with improved creep properties.     

Novel Carboxylic Acid Solvolysis Route to Synthesis Electrode-active Nanostructured Spinel-Li_4Ti_5O_(12) for Lithium Batteries and Hybrid Capacitors

1 Results Nanostructured spinel-type Li4Ti5O12 (LTO) was prepared using Lewis acid base reaction technique involving a mixture of titanium β-diketonate and lithium nitrate as starting materials in the presence of aqueous citric acid as a solvolysis agent. The above method yielded a simple single step process without involving sol to gel conversion. The phase purity of the synthesized product after calcining at 800 ℃ for 24 h in air showed a spinel structure without any residual impurities. The nanostructure of the final product (both as prepared and calcined) was confirmed by High-resolution TEM (HRTEM) analysis. This process yielded very uniform spherical nanoparticles of ≈3 nm in size. The LTO product was tested as anode-active material in lithium containing half cells, Li4Ti5O12/Li at room temperature. It was found that the composite anode exhibited a working voltage of 1.55 V vs. Li/Li with a first discharge capacity of 160 mAh·g-1.