Reversible 5′-end biotinylation and affinity purification of synthetic RNA

A reversible biotinylation phosphoramidite was synthesized and incorporated onto the 5′-end of an oligoribonucleotide on a solid phase synthesizer. After cleavage and deprotection, the crude synthetic oligomer mixture was incubated with NeutrAvidin^® coated microspheres, and the failure sequences removed by washing with a buffer followed by treating the microspheres with tetrabutylammonium fluoride to give a high quality unmodified full-length oligoribonucleotide.     

Fractionation and purification of antioxidant peptides from Chinese sturgeon (Acipenser sinensis) protein hydrolysates prepared using papain and alcalase 2.4L

Protein hydrolysates have the potential to be natural and safer sources of bioactive peptides. In this study, two proteases were used to hydrolyze Chinese sturgeon (Acipenser sinensis) protein, and the hydrolysates were then purified to yield antioxidant peptides. The degree of hydrolysis of 23.56 % and 18.14 % was obtained using papain and alcalase 2.4L, respectivly, and hydrolysates had 96.80 % and 87.24 % total amino acid content, respectivly. The papain hydrolysate (PH) and alcalase 2.4L hydrolysate (AH) showed good antioxidant activity against DPPH^? (IC₅₀ of 3.64 and 3.15 mg/mL) and ABTS^?+ (IC₅₀ of 1.92 and 1.58 mg/mL), respectively. The low-molecular-weight (<1000 Da) fraction of both hydrolysates demonstrated the highest antiradical activity (IC₅₀ of 2.59 and 2.31 mg/mL, DPPH) and (IC₅₀ of 1.54 and 1.36 mg/mL, ABTS), respectively. Nine peptides were separated from both hydrolysates using reverse phase high performance liquid chromatography (RP-HPLC). The IC₅₀ for ABTS^?+ scavenging activity of peptide P5 with valine, glycine and asparagine (MW of 282.13 Da) from PH, and peptide P3 with histidine, glycine and alanine (MW of 302.74 Da) from AH was 0.89 and 0.72 mg/mL, respectively. The fractions and purified peptides obtained from Chinese sturgeon hydrolysates could be utilized as natural antioxidant substitutes in pharmaceuticals and food products.     

Production and Purification of Pectinase from Bacillus subtilis 15A-B92 and Its Biotechnological Applications

Enzymes that degrade pectin are called pectinases. Pectinases of microbial origin are used in juice clarification as the process is cost-effective. This study screened a pectinase-producing bacterium isolated from soil and identified as Bacillus subtilis 15A B-92 based on the 16S rRNA molecular technique. The purified pectinase from the isolate showed 99.6 U/mg specific activity and 11.6-fold purity. The molecular weight of the purified bacterial pectinase was 14.41 ± 1 kD. Optimum pectinase activity was found at pH 4.5 and 50 °C, and the enzyme was 100% stable for 3.5 h in these conditions. No enzymatic inhibition or activation effect was seen with Fe²⁺, Ca²⁺, or Mg²⁺. However, a slight inhibition was seen with Cu²⁺, Mn²⁺, and Zn²⁺. Tween 20 and 80 slightly inhibited the pectinase, whereas iodoacetic acid (IAA), ethylenediaminetetraacetate (EDTA), urea, and sodium dodecyl sulfate (SDS) showed potent inhibition. The bacterial pectinase degraded citrus pectin (100%); however, it was inactive in the presence of galactose. With citrus pectin as the substrate, the Km and Vmax were calculated as 1.72 mg/mL and 1609 U/g, respectively. The high affinity of pectinase for its substrate makes the process cost-effective when utilized in food industries. The obtained pectinase was able to clarify orange and apple juices, justifying its application in the food industry.     

HCVNS2基因表达载体的构建及在E.coli中表达

PCR人含有丙肝病毒全长非结构蛋白的载体pBlueBac25中扩增出全长的NS2基因DNA片段,分别克隆到表达载体pQE30和转座载体pFasBacHTb的多隆位点（MCS）,PFastNS2通过转座插入穿梭载体Bacmid的表达盒;pQENS2转化JM109菌株,诱导表达出N端含有6个His的全长His的全长NS2蛋白,用Ni-NTA-agarose柱层纯化,获得提纯的全长NS2蛋白。     

Uptake of Europium(III) from Water using Magnetite Nanoparticles

It is demonstrated that colloidal magnetite nanoparticles can be used as nanosorbents for lanthanide ions dissolved in water. In particular, a series of experiments are performed for the removal of Eu(III) in distinct analytical conditions and by applying an external magnet to collect the sorbents previously dispersed in water samples. Furthermore, strategies for surface chemistry functionalization are also investigated, aiming to investigate the effect of this parameter on the removal capacity of the Fe₃O₄ nanoparticles. The supernatant solutions are monitored for the remaining amount of Eu(III) by fluorescence emission measurements in the presence of 2,6‐pyridinedicarboxylic acid as a sensitizer. The results demonstrate that neat Fe₃O₄ nanoparticles are capable of capturing lanthanide ions (III) from aqueous solutions (pH 7), without need of surface modification, and for subsequent removal by magnetic separation. During the removal, efficiency is increased after modifying the particles' surfaces with silica and 3‐aminopropyltrimethoxysilane; in alkaline medium (pH 10), there is complete removal regardless the type of nanosorbent used. This has been explained by the formation of insoluble Eu(III) species that adsorb strongly to the nanosorbents surfaces allowing their subsequent magnetic separation.     

Influence of oxygen on the ion-assisted regrowth of deposited Si layers

Summary The ion-assisted regrowth of chemical-vapour deposited Si films onto (100) Si substrates is reported. The regrowth was induced by a 600 keV Kr⁺⁺ beam at doses in the range (2·10¹⁵÷6·10¹⁵)/cm² and at a dose rate of 1·10¹²/cm²s. The target temperature was set at 450°C. During irradiations the crystal-amorphous interface velocity was measuredin situ by monitoring the reflectivity of a He-Ne laser light focused onto the sample surface. After irradiation some samples were also analysed by Rutherford backscattering in combination with the channelling effect and by transmission electron microscopy. The growth rate of deposited layers depends on the cleaning procedure performed prior to deposition,i.e on the total amount of oxygen present at the deposited layer/substrate interface. Moreover, twinned material is observed in the recrystallized layers and its concentration is strongly dependent on substrate cleaning. These phenomena are explained in terms of a decrease in the ion-assisted growth rate in the presence of high oxygen concentrations. The data are discussed and compared with those obtained during pure thermal annealing. To speed up publication, the authors of this paper has agreed to not receive the proofs for correction.     

大孔树脂分离纯化红旱莲黄酮

通过静态吸附和解吸实验,筛选适合分离纯化红旱莲黄酮的大孔吸附树脂并确立纯化工艺参数。结果表明,NKA-9型大孔吸附树脂是性能良好的红旱莲黄酮吸附剂,其最佳吸附条件为样液初始浓度3.5mg.mL-1,样液pH 6.0,吸附温度35℃;最佳解吸条件为解吸乙醇体积分数60%,解吸液料比101(60%乙醇溶液大孔吸附树脂,mL.g-1)。     

Toward Uniform Optical Properties of Carbon Dots

Carbon dots possess versatile optical properties that have prompted their investigation in applications including photocatalysis, photovoltaics, imaging, and drug delivery, among others. However, the preparation of these nanodots is accompanied by the formation of fluorophores and intermediates, which can be difficult to separate. In the absence of thorough purification protocols, the reported optical properties are often heterogeneous, which hinders understanding of their physicochemical and optical properties and concrete application development. Here, two hydrophilic carbon dot systems starting with citric acid and diethylenetriamine are prepared. The impact of purification, including dialysis, ultrafiltration, and organic washes, on the properties of the dots is demonstrated. It is shown that monitoring the purification endpoint using near-infrared, fluorescence, and absorbance spectroscopies is possible. Moreover, it is demonstrated that fluorescence quantum yields can be a reliable tool to determine the purification endpoint. This work shows that even carbon dots derived from the same chemical precursors can have different purification profiles and purification requirements. However, the developed approach can be used to determine the proper purification procedure and endpoint for any carbon dot system regardless of the starting materials. Finally, it is envisioned that this work can be easily extended toward the purification of other hydrophilic nanomaterials.