Electrosome

Electrosome

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  Electrosomes Mr. Dighe AjinkyaD M- Pharm 1st year, Sem II (Pharmaceutics) Guided By - Ms. Bidkar Mam Presented At- Sharadchandra Pawar Collage Of Pharmacy Otur
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  Contents 1. Introduction 2. Methodof preparation 3. Advantages Electrosomes 4. Disadvantages Electrosomes 5. Application 6. Reference
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  INTRODUCTION • These arethe transmembrane protein generate and propagate the electrical signals that allow us to sense our surroundings, process, information, make decisions, and move. • Ion channel proteins act as gates that span the lipid bilayer that surrounds all electrochemical gradients. • The ion flux through a channel pore can be extremely high. • They are high resolution in function and 3D structure to description of their molecules. • The high resolution structure of ion channel and ion channel associated protein are providing the substrates for sophisticated tests of the mechanisms of channel gating and permeation. • Ion perform two basic function open and close to control the passage of ion across the cell membrane.
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  Extracellular Activation signal (voltage charge,ligand Binding, second messenger) Cytoplasm
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  • The electrosomes,a novel surface-display system based on the specific interaction between the cellulosomal scaffoldin protein and a cascade of redox enzymes that allows multiple electron-release by fuel oxidation. • The electrosomes is composed of two compartment: (i) a hybrid anode, which consists of dockerin-containing enzymes attached specifically to cohesin sites in the scaffoldin to assemble an ethanol oxidation cascade, and (ii) a hybrid cathode, which consists of a dockerin-containing oxygen- reducing enzyme attached in multiple copies to the cohesin-bearing scaffoldin.
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  • The electrosomeswas designed for use both in an anode and a cathode compartment; in each compartment, the unique attributes of the cellulosome scaffoldin give a different advantage • In the anode, the ethanol oxidation cascade consists of two enzymes, ADH and formaldehyde dehydrogenase (FormDH), both containing a different dockerin module of Acetivibrio cellulolyticus and of Clostridium thermocellum, C. thermocellum (zADH-Ac and pFormDH-Ct), respectively, assembled on a ‘designer’-scaffoldin chimera displayed on the surface of S. cerevisiae. • At the cathode, copper oxidase (CueO) was selected for surface-display. CueO is a multi- copper oxidase enzyme expressed by E. coli that catalyzes the oxidation of Cu(I) ions coupled to oxygen reduction to water. • The different constructs used for assembly are depicted. We report the characterization of the dockerin-containing enzymes and their electrochemical activity using a diffusing redox mediator.
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  Method of Preparation 1.Strains and Constructs method. 2. Enzyme Binding to Scaffoldin. 3. Biofuel-Cell Assembly and Characterization. 4. Protein Expression. 5. Enzyme Activity Assays. 6. Construction of YSD of Chimeric Scaffoldins. 7. Cyclic Voltammetry (CV) and Chronoamperometry (CA).
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  Strains and Constructsmethod • The genes encoding dockerins of Acetivibrio cellulolyticus and Clostridium thermocellum were cloned and ligated to the C-terminus of Zymomonas mobilis alcohol dehydrogenase and to Pseudomonasputida formaldehyde dehydrogenase by standard methods. • The dockerin module of C.thermocellum was also ligated to the C-terminus of CueO (CueO- Ct) of E.coli. • All the dockerin-containing enzymes encoding genes have been cloned into the pET15b vector for expression in E. coli, yielding the pET15b-zADH-Ac, pET15b- pFormDH-Ct, and pET15b-CueO-Ct vectors. • For controls, the genes encoding the native enzymes without an appended dockerin module were also cloned in the same vector, yielding plasmids pET15b- zADH, pET15b-pFormDH, and pET15b-CueO.
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  Enzyme Binding toScaffoldin • 2.0 mL of yeast cells displaying scaffoldin, for which absorbance at a wavelength of 600 nm was 1.0 , were incubated with bacterial lysates containing the expressed enzymes at room temperature for 1 h. 1.0 mL of the bacterial lysates were used for the binding, which was performed in a final volume of 15 ml. • As a binding buffer, 50 mM Tris buffer at pH 8.0 with 1 mM CaCl2 was used. Upon binding, the yeast cells were precipitated, and binding was repeated using fresh lysate. • After the second binding cycle, the yeast cells were washed four times in the buffer to remove non-specifically bound enzymes. • For the CueO-Ct binding, the yeast cells were suspended in 0.1m acetate buffer pH 5.0 containing 1 mm CaCl2 after the last wash. • Following binding, the yeast cells were resuspended in 2.0 ml of buffer
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  Biofuel-Cell Assembly andCharacterization • Air was continuously purged to the fuel-cells. A potentiostatically controlled anode set to −0.2 V versus Ag/AgCl was used. • In all experiments, the cells were left to stabilize overnight, following fuel cell assembly, before characterization was performed. • The characterization of fuel cell performance was done by measuring the voltage of the cells under variable external loads. • A background current cell was used as a negative control for all fuel cell experiments and did not contain any yeast. Graphite rods of 5 mm diameter served as both anodes and cathodes. • The counter electrode that served for the potentiostatically controlled electrode was of a larger surface area, as described for the CV and CA measurements
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  Advantages • It perpetuatesthe endurance of active drug molecule in the systemic circulation. Deferment the elimination reactions of promptly metabolize drugs and contributes to controlled release. • Incorporates both hydrophilic and lipophilic drugs. • Intensifies the stability of medicament. • Cost of therapy is minimized by reducing the dose per unit formulation • Elevate bioavailability especially in water disfavouring drugs. • Selective uptake by tissues due to direct drug delivery.
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  Disadvantages • The productioncost of electrosomes are generally high since these come under the class of nanotherapeutics. • The constituent phospholipids present in lipid vesicular structures may undergo oxidation or hydrolysis.
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  Application • They useenzymatic reactions to catalyze the conversion of chemical energy to electricity in a fuel cell. • The use of enzymatic cascades in enzymatic fuel cell anodes resulted in very high power outputs, as the electron density achieved was much higher when the fuel was fully oxidized. • Its used as a carrier in drug targeting. • Used in the treatment of cancer. • Used in studying immune response. • Ear targeting • Muscle targeting
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  Reference 1. SHEFRIN S,SREELAXMI C. S, VISHNU VIJAYAN, SREEJA C. NAIR.ENZYMOSOMES: A RISING EFFECTUAL TOOL FOR TARGETED DRUG DELIVERY SYSTEM.INT J APP PHARM. 2017;9(6);1-9 2. 2. SZCZUPAK A, AIZIK D, MORAÏS S, VAZANA Y, BARAK Y, BAYER A E, ALFONTA L. THE ELECTROSOME: A SURFACE-DISPLAYED ENZYMATIC CASCADE IN A BIOFUEL CELL’S ANODE AND A HIGH-DENSITY SURFACE-DISPLAYED BIOCATHODIC ENZYME.NANO- MATERIAL.2017 3. 3. WWW.SCIENCE DIRECT.COM