Solvent-induced 7R-dioxygenase activity of soybean 15-lipoxygenase-1 in the formation of omega-3 DPA-derived resolvin analogs

The resolvin family contains important anti-inflammatory and pro-resolution compounds enzymatically derived in vivo from the polyunsaturated omega-3 fatty acids, eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA). More recently, docosapentaenoic acid (DPA) has emerged as another potentially important precursor in the biological production of resolvin compounds. In this work we have used medium engineering to develop a simple method for the controlled synthesis of two di-hydroxylated diastereomers of DPAn-3 catalyzed by soybean 15-lipoxygenase-1 (15-sLOX-1) in the presence of short chain n-alcohols, including methanol, ethanol and propan-1-ol. The complete structures of the two major products, 7S,17S-dihydroxydocosapenta-8Z,10E,13Z,15E,19Z-enoic acid (7S,17S-diHDPAn-3) and 7R,17S-dihydroxydocosapenta-8Z,10E,13Z,15E,19Z- enoic acid (7R,17S-diHDPAn-3), have been elucidated using spectroscopic analysis. The alcohol-dependent R-dioxygenase activity of soybean 15-lipoxygenase with mono-hydroperoxide intermediate substrates has also been demonstrated with other biologically relevant PUFAs, including DHA, EPA and ARA. The developed method has applications in the production of closely related isomers of naturally occurring resolvins and protectins, demonstrating the versatility of 15-sLOX-1 as a biocatalyst. © 2014 Elsevier B.V.

Graphene nanodots encaged 3-D gold substrate as enzyme loading platform for the fabrication of high performance biosensors

Herein, a uniform three-dimensional (3-D) graphene nanodots-encaged porous gold electrode was prepared via ion beam sputtering deposition (IBSD) and mild corrosion chemistry for efficient enzyme electrode fabrication. Enzymes, like glucose oxidase and catalase, were modified with pyrene functionalities and then loaded into the graphene nanodots encaged porous gold electrode via non-covalent π-π stacking interaction between pyrene and graphene. The fabricated enzyme electrodes showed profound reusability and repeatability, high sensitivity, inherent selectivity and enhanced detection range. As for glucose analysis a broad linear range from 0.05 to 100 mM was obtained and the linear range for hydrogen peroxide was 0.005 to 4 mM. Detection limits of 30 μM for glucose and 1 μM for hydrogen peroxide were achieved (S/N = 3), respectively. These electrodes can be applied to analyze the clinical samples with reliable results. The formation mechanism and 3-D structure of the porous electrode were investigated using high resolution transmission electron microscope (HRTEM), atomic force microscopy (AFM), scanning electron microscope (SEM) and electrochemical impedance spectroscopy (EIS). Most importantly, various other ideal biosensors can be fabricated using the same porous electrode and the same enzyme modification methodology.