The document discusses artificial enzymes, synthetic molecules that mimic natural enzymes' functions, including their properties, need, design approaches, ideal environments, and applications. It highlights the advantages of artificial enzymes like high catalytic efficiency, tunable structures, and applicability in pharmaceuticals and genetic engineering. The document also covers methods to design these enzymes, emphasizing chemical and biological approaches, and outlines specific examples such as xnazymes and manganese dioxide nanozymes.

1. AMITY INSTITUTE OF PHARMACY
ARTIFICIAL ENZYMES
Submitted to: Submitted by:
Dr. Viney Lather Firuj Ahmed
Department of Pharmaceutical Chemistry Master of Pharmacy
1st Year

2. CONTENTS:
1. Enzymes
2. Properties of Enzymes
3. Artificial Enzymes
4. Ideal characteristics of Artificial Enzymes
5. Need of artificial Enzymes
6. Designed approach for Artificial Enzymes
7. Ideal environment requirement for Artificial Enzymes
8. Applications.

3. ENZYMES
• Enzymes are biological catalyst
• A catalyst is defined as “a substance that increases the rate of a chemical
reaction without being itself changed in the process.”
Enzymes as biological catalysts:
• Enzymes are proteins that increase the rate of a reaction by lowering the
activation energy.
• They catalyzed nearly all the reactions taking place in the cells of the
body.
• Enzymes have unique three-dimensional shapes that fit the shapes of
substrates.

4. Properties of enzymes
• Catalytic efficiency: High efficiency, 103 to 1017 faster than the
corresponding catalyzed reaction.
• Specificity: High specific in nature, interacting with one or a few
specific substrates and catalyzing only one type of chemical reaction.
• Mild reaction condition:
 Temperature: Optimum temperature of enzymes is 20-35 degree
Celsius. They become inactivated at very low temperatures and
denatured at very high temperature( greater than 45 degree Celsius).
 pH: Most enzymes exhibit optimal activity at a pH value between 5 and
9. High or low pH value than the optimum value will cause ionization of
enzyme which results in the denaturation of the enzyme.

5. ARTIFICIAL ENZYMES
 An artificial enzyme is a synthetic organic molecule or ion that mimics one or more
functions of an enzyme.
 Molecules are designed and modified to achieve some desirable features of
enzymes.
 Protein engineering has been developed to design and synthesize molecules with
the attributes of enzymes for non-natural reactions.
 They have a molecular weight of less than 2000 Dalton.
 They have the ability to stabilize at a higher temperature.
 They are also known as synzymes or enzyme mimics.
 Nanozymes: Nanozymes are nanomaterials with enzyme-like characteristics.
They have been explored for applications such as biosensing, bioimaging, tumor
diagnosis and therapy, and anti-biofouling, etc.

6. CHARACTERISTICS OF ARTIFICIAL ENZYMES
 Rigid structure model.
 Water soluble and catalytically active under physiological
conditions.
 Reversible, non-covalent binding with the substrate, the release
is slower than the binding.

7. EXAMPLES:
 XNAzymes: These XNAzymes are capable of cutting and joining strands
of RNA in a test tube. One of the XNAzymes are much more stable than
naturally occurring enzymes, scientist believes that they could be
particularly useful in developing new therapies for a range of disease,
including cancer and viral infection. XNAzymes are chemically extremely
robust and because they do not occur in nature and are not recognized by
the body’s natural degrading enzymes. This might make them an
attractive candidate for the long-lasting treatment that can disrupt
disease-related RNAs.
 Manganese dioxide nanozymes(Mn3O4) ROS(Reactive Oxygen Species)
scavenging activities have been developed for in vivo anti-inflammation.
 Vanadium pentoxide mimics to glutathione peroxidase.

8. NEED OF ARTIFICIAL ENZYMES
 Tunable structures and catalytic efficiencies are similar to the natural
enzyme.
 Excellent tolerance to experimental conditions.
 Purely synthetic routes for their preparation.
 High cost and low stability limit the application of natural enzymes.
 Speeds up the reaction at a relatively high rate.

9. DESIGN APPROACH FOR ARTIFICIAL ENZYMES
Two types of approaches are used to develop artificial enzymes:
1. Chemical Approach:
 Cyclodextrins as enzyme mimics
 Cyclophane as enzyme mimics
 Calixarene as enzyme mimics
 Crown ethers as enzyme mimics
2. Biological Approach:
 Direct evolution method for artificial enzyme production
 De novo computational Enzyme Designing

10. CHEMICAL APPROACH
Cyclodextrins:
 The three most common
cyclodextrins are , α- ,β-,and γ-
species, which are composed of
six, seven, and eight
glucopyranose units, respectively.
Hydrophobic cavity.
 Stable and water soluble.
 Tunable (modify to change
properties)
 Research on cyclodextrin started
in1930s very expensive &thought
to be toxic

11. DIRECT EVOLUTION METHOD FOR ARTIFICIAL ENZYME
PRODUCTION
 Direct evolution is a molecular biology method to modify biocatalysts via in vitro version of
“Darwinian evolution”.
 Direct evolution provides improved enzymatic activity, thermostability, tolerance to organic
solvents, substrate specificity, enantioselectivity, and so on.
Gene mutagenesis insertion of a gene sequence into host cells transformed colony planting
Evaluated by using screening for transformed Bacteria producing
enzymatic assays colonies mutant enzymes
More production of
Transformed colonies

12. De novo computational enzyme designing
 Under this automated identification of amino acid sequences performed that
will fold into a specified three-dimensional structures. This method has
emerged as a promising tool for engineering enzymes .
 Conformational changes are part of the repertoire that natural enzymes use
to catalyze reactions.
 In order to select candidates sequences for enzymes at each step of the
identified pathways, several tools provide different solutions, including anti
SMASH for biosynthetic gene clusters.
 Tools used:
 Path pred
 RDM patterns
 Bond-Electron matrices
 Reactions SMARTS in Retro
 Path 2.0

13. Ideal Environment Requirement for
Artificial Enzymes
 The covalent immobilization of one enzyme on solid supports having large
surfaces with given properties (eg., hydrophilic or hydrophobic surfaces) should
provide a nano environment surrounding the area of the enzyme directly in
contact with the support.
 Further covalent immobilization of macromolecular polymers (hydrophilic,
hydrophobic) on the same large internal surfaces of the solid support should
provide an additional nano environment surrounding the area of the
immobilized enzyme molecules next to the support.
 Chemical modification of immobilized enzymes with polyfunctional
macromolecules could also be an interesting way to greatly modified the
enzyme nano environment with minimal chemical modification of the enzyme.

14. APPLICATIONS
 Tunable structure and catalytic efficiencies similar to natural enzymes.
 Excellent tolerance to experimental conditions.
 Purely synthetic routes for their preparation.
 Lower cost.
Pharmaceutical : synthetic enzymes that accelerates the formation of
drugs and chemicals.
Medicine : use of synthetic enzymes as supplements for patients deficient
in certain enzyme can be made instead of extracting natural enzymes from
other organisms.
Genetic Engineering : potentially designing synthetic enzymes that
manipulate gene sequences to create genetically modified organisms or to
help genealogy research.

15. THANK YOU