Development of Batch and Continuous Processes on Biodiesel Production in a Packed-Bed Reactor by a Mixture of Immobilized Candida rugosa and Rhizopus oryzae Lipases

In this study, transesterification and esterification were investigated in batch and continuous process using immobilized Candida rugosa and Rhizopus oryzae lipases. In the case of batch process, stepwise reaction method was investigated to prevent the lipase deactivation. Reaction conditions were as follows: temperature, 45 °C; agitation speed, 250 rpm; enzyme concentration, 20%; and water contents 10%. And then, conversion yield was 98.33% at 4 h. In the case of continuous process, circulation and long-term continuous system were investigated for development of efficient mass transfer system. Optimal reaction conditions were as follows: temperature, 45 °C; flow rate, 0.8 mL/min; and water contents, 10%. And then, conversion yield of biodiesel was 97.98% at 3 h. Especially, the maximum conversion yield using a mixture of immobilized lipases exceeded over 90% for 108 h in long-term continuous system under optimal reaction conditions (45 °C; flow rate, 0.8 mL/min; and water contents, 10%). These results should help in determining the best method for the biodiesel production and improving the design and operation of large scale by enzymatic systems.     

Electrospinning fabrication and characterization of poly(vinyl alcohol)/montmorillonite/silver hybrid nanofibers for antibacterial applications

Poly(vinyl alcohol) (PVA)/montmorillonite clay (MMT)/silver (Ag) nanoparticles have been electrospun for fabricating PVA/MMT/Ag nanofiber in aqueous solutions. Since PVA is a water-soluble and biocompatible polymer, it is one of the best materials for preparation of antibacterial nanofiber. MMT has been used as an inorganic filler to enhance properties of homopolymeric nanofiber. The PVA/MMT/Ag nanofiber diameter increases with increasing contents of MMT clay and Ag nanoparticles. In preservation test, the PVA/MMT/Ag nanofiber confirms an excellent antibacterial performance, elucidating for practical uses as a new preservative. Moreover, the PVA/MMT/Ag nanofiber shows improved thermal properties.     

Selective inhibitory effect of epigallocatechin-3-gallate on migration of vascular smooth muscle cells

In order to prevent restenosis after angioplasty or stenting, one of the most popular targets is suppression of the abnormal growth and excess migration of vascular smooth muscle cells (VSMCs) with drugs. However, the drugs also adversely affect vascular endothelial cells (VECs), leading to the induction of late thrombosis. We have investigated the effect of epigallocatechin-3-gallate (EGCG) on the proliferation and migration of VECs and VSMCs. Both cells showed dose-dependent decrease of viability in response to EGCG while they have different IC(50) values of EGCG (VECs, 150 mM and VSMCs, 1050 mM). Incubating both cells with EGCG resulted in significant reduction in cell proliferation irrespective of cell type. The proliferation of VECs were greater affected than that of VSMCs at the same concentrations of EGCG. EGCG exerted differential migration-inhibitory activity in VECs vs. VSMCs. The migration of VECs was not attenuated by 200 mM EGCG, but that of VSMCs was significantly inhibited at the same concentration of EGCG. It is suggested that that EGCG can be effectively used as an efficient drug for vascular diseases or stents due to its selective activity, completely suppressing the proliferation and migration of VSMCs, but not adversely affecting VECs migration in blood vessels.     

Electrochemical performances of inorganic membrane coated electrodes for li-ion batteries

In the present study, we prepared new Li-ion batteries with inorganic membrane-coated electrodes fabricated by the direct coating of a 400-nm-sized Al₂O₃ particles onto the anodes and studied their charge–discharge characteristics for the first time. Being made of 94 wt.% Al₂O₃ powder and 6 wt.% polymeric binder, the membranes on the anodes showed excellent wettability with the liquid electrolytes, due to their high porosity and capillarity. The Li-ion batteries prepared by the assembly of the inorganic membrane-coated electrodes showed excellent charge–discharge behavior compared to conventional Li-ion batteries with a polymeric separator. However, their high-rate performance was affected by the binder contents in the inorganic membranes. The thermal properties of the Li-ion battery with the inorganic membrane-coated electrodes were compared with those of a conventional one with a polymeric separator.     

Microchip electrophoretic separation for the fast diagnosis of Anaplasma phagocytophilum infection in cattle

We report a diagnostic method for Anaplasma phagocytophilum (A. phagocytophilum) infection in cattle using a nested PCR and microchip electrophoresis (ME). A. phagocytophilum causes human granulocytic anaplasmosis and granulocytic ehrlichiosis, which are emerging tick‐borne zoonotic diseases. Nested PCR was used to amplify genomic DNA samples extracted from cattle blood. The amplified PCR products were analyzed under a sieving gel matrix of 0.7% poly(ethyleneoxide) (M_r=8 000 000) in a conventional glass microchip. In the ME assay, A. phagocytophilum was analyzed within 35 s with a relative standard deviation of 1.30% (n=5) using a programmed field strength gradient (PFSG) as follows: 615.3 V/cm for 0–24 s, 66.7 V/cm for 24–34 s, 615.3 V/cm for 34–100 s. The ME‐PFSG assay was clinically validated by comparing the 16S rRNA gene levels obtained by this method with those measured using conventional slab gel electrophoresis performed with ten cattle blood samples suspected of A. phagocytophilum infection. In contrast to slab gel electrophoresis, the proposed ME‐PFSG methodology had increased sensitivity (200–450 pg/μL), a faster analysis time (<35 s), and required a smaller sample volume (～162 fL).     

Enzymatic transformation of platycosides and one‐step separation of platycodin D by high‐speed countercurrent chromatography

Platycosides, the saponins found in the roots of Platycodon grandiflorum (Platycodi Radix), are typically composed of oleanane triterpenes with two side chains. In platycosides, platycodin D, a glucose unit at C‐3, is a major component, which has several pharmacological activities. Because of the high demand for this compound, we attempted to enzymatically convert platycodin D₃ and platycoside E, having two and three glucose units at C‐3, respectively, into platycodin D. In this study, we tested the ability of several glycosidases to transform platycosides, or more specifically, the ability to transform platycoside E and platycodin D₃ into platycodin D. To obtain pure platycodin D on a preparative scale, high‐speed countercurrent chromatography with a solvent system of ethyl acetate/n‐butanol/water (1.2:1:2, v/v/v) was used for the separation of the enzymatically transformed product. Approximately 39.4 mg of platycodin D (99.8% purity) was obtained from 200 mg of the product in a one‐step separation. The results strongly support the advantage of enzymatic transformation of the platycosides for the efficient enrichment of platycodin D in the complicated extract of the medicinal plant.     

Sensitivity enhancement of CE and CE-MS for the analysis of peptides by a dynamic pH junction

An analytical method of CE-MS and CE with an online preconcentration technique induced by a dynamic pH junction, addition of organic solvent and large volume injection was developed for sensitive determination of peptides in biological samples. Leucine enkephalin, methionine enkephalin, dynorphin A, β-endorphin and angiotensin II were used as model peptides. The optimal online preconcentration conditions were obtained at a sample matrix consisting of 100?mM borate buffer (pH 10.0) with 50% v/v acetonitrile and a BGE containing 1?M formic acid at pH 2.0, along with a 25-cm injection length. Under the optimized conditions, a 4.0×10(3)-1.1×10(4)-fold increase in peak intensity was achieved without degrading the peak shape. This online preconcentration method was applied to analyze the intracellular angiotensin II within the peptides extracted from HL1 cells and approximately increase of 1×10(4)-fold sensitivity was achieved compared to normal condition. Thus, the developed method could be applied to the analysis of various peptides for peptidomics study in biological samples.