This document describes electrosomes, which are a novel surface display system composed of two compartments - a hybrid anode and cathode. The anode uses a scaffolding protein to assemble an ethanol oxidation enzyme cascade on the surface of Saccharomyces cerevisiae. The cathode similarly uses a scaffolding protein to display multiple copies of a oxygen-reducing enzyme. Electrosomes were designed for use in both compartments to catalyze the conversion of chemical energy to electricity in a fuel cell. They allow high electron density and power output. The document discusses their preparation, advantages of controlled drug release and targeting applications, and disadvantages related to production costs.

1. COPS DSU
DEPARTMENT OF PHARMACEUTICS
PRESENTED BY:
SAGAR.G
M.PHARM,2nd SEM
DEPARTMENT OF PHARMACEUTICS
SUBMITTED TO :
Dr.B. WILSON
HEAD OF THE DEPARTMENT
DEPARTMENT OF PHARMACEUTICS
ELECTROSOMES

2. COPS DSU
DEPARTMENT OF PHARMACEUTICS
CONTENTS
INTRODUCTION
METHOD OF PREPARATION
ADVANTAGES OF ELECTROSOMES
DISADVANTAGES OF ELECTROSOMES
APPLICATION
REFERENCE

3. COPS DSU
DEPARTMENT OF PHARMACEUTICS
INTRODUCTION
• These are the 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.

4. COPS DSU
DEPARTMENT OF PHARMACEUTICS
Extracellular
Activation signal
(voltage charges, ligand binding, second
messenger)
Cytoplasm

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DEPARTMENT OF PHARMACEUTICS
• 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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DEPARTMENT OF PHARMACEUTICS
• The electrosomes was 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 catalyses 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.

7. COPS DSU
DEPARTMENT OF PHARMACEUTICS
+
TARGET
PROTEIN
NANOCARRIER
DIFFERENT
LOADING
TECHNIQUES
DIRECT
CONJUGATION
METHOD
PHYSICAL
ABSORPTION
METHOD
ENCAPSUL
ATION

8. COPS DSU
DEPARTMENT OF PHARMACEUTICS
Structure of ELECTROSOMES
Ion channels are the part of large protein network. These networks includes
cytoskeletonComponents , singling proteins like protein kinase and phosphates, and channel-
associated proteins that recruit singling molecules to the channel to modify its function.
Gold nanoparticle
Physicochemical properties:
Shape, size, surface coating
Etc.
Experimental condition:
Administration route, dose,
Animal species, exposure duration, etc.
Pharmacokinetics and biodistribution
Tumor targeting (EPR effect)
Liver uptake (RES)
Penetration of biological barriers
(eg.BBB)
Renal clearance
Metabolism
Biological effects
Therapeutic efficacy
Toxicity {
Cytotoxicity
In vivo toxicity

9. COPS DSU
DEPARTMENT OF PHARMACEUTICS
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)

10. COPS DSU
DEPARTMENT OF PHARMACEUTICS
• 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.
Strains and Constructs method

11. COPS DSU
DEPARTMENT OF PHARMACEUTICS
• 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, they east cells were resuspended in 2.0ml of buffer
Enzyme Binding to Scaffoldin

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DEPARTMENT OF PHARMACEUTICS
• 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 stabilise overnight, following
fuel cell assembly, before characterization was performed.
• Thecharacterizationoffuelcellperformancewasdonebymeasuringthevolta
geofthecells 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
Biofuel-Cell Assembly and Characterization

13. COPS DSU
DEPARTMENT OF PHARMACEUTICS
ADVANTAGES OF ELECTROSOMES
• It perpetuates the endurance of active drug molecule in the systemic
circulation. Deferment the elimination reactions of promptly metabolise
drugs and contributes to controlled release.
• Incorporates both hydrophilic and lipophilic drugs.
• Intensifies the stability of medicament.
• Cost of therapy is minimised by reducing the dose per unit formulation
• Elevate bioavailability especially in water disfavouring drugs.
• Selective uptake by tissues due to direct drug delivery.

14. COPS DSU
DEPARTMENT OF PHARMACEUTICS
DISADVANTAGES OF ELECTROSOMES
• The production cost 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.

15. COPS DSU
DEPARTMENT OF PHARMACEUTICS
APPLICATION
• They use enzymatic reactions to catalyse 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 oxidised.
• It's used as a carrier in drug targeting.
• Used in the treatment of cancer.
• Used in studying immune response.
• Ear targeting
• Muscle targeting

16. COPS DSU
DEPARTMENT OF PHARMACEUTICS
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. 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. WWW.SCIENCE DIRECT.COM

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DEPARTMENT OF PHARMACEUTICS
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