CHEMISTRY OF NATURAL

PRODUCTS

B.Pharm PCG302

Dr. Ibrahim Malami

Department of Pharmacognosy & Ethnopharmacy, Faculty of
Pharmaceutical Sciences, Usmanu Danfodiyo University, Sokoto.

Centre For Advanced Medical Research and Training,

Usmanu Danfodiyo University, Sokoto.
PHYTOCHEMISTRY OF ENZYMES AND
ENZYMES CONTAINING DRUGS

(2 hours)
 Objectives

At the end of this set of lectures, you should:

 Be able to understand the properties of enzymes, their

chemical nature, classification and pharmaceutical uses.
 Be able to identify drugs containing enzymes in any
community pharmacy.

 Be able to understand their method of preparation.

Highlight:

 Introduction
 Classification and nomenclature of enzymes
 Coenzymes
 Industrial applications
 Natural digestive enzymes
 papain
 bromelain
Introduction

 Enzymes – protein molecules that act as catalyst to carry out

multiple metabolic activities.

 They breakdown complex molecules – proteins, fats and

carbohydrates by the action of enzymes into simple
substances that can easily be excreted out as byproduct.

 Enzyme – is a biological or organic catalyst which can alter or

speed up a chemical reaction, without itself being chemically
change at the end of the chemical reaction.
Sources
Enzymes – extracted from any living organism including bacteria,
fungi, yeast, plants and animals.
Importance of enzymes

Breakdown:
Enzymes – breakdown complex substances to simpler
substances, for example the oxidation of glucose to release
energy and form carbon dioxide and water.
Synthesis
Other – synthesize complex substances from simpler ones, for
example the build up of proteins from amino acids.
Classification and nomenclature of enzymes :
Traditionally, enzymes were simply assigned names by the
investigator who discovered the enzyme e.g. zymase, permease,
translocase, replicase, etc.

Knowledge expanded – comprehensive and complex.

Most enzymes are named after the reaction they catalyze and
the name ends in ase e.g. Hexokinase, DNA polymerase etc.

Currently – six functional classes of enzyme by the International

Union of Biochemists (I.U.B.).
 Classification Biochemical properties

1 Oxidoreductases Act on many chemical groupings to add or remove hydrogen

GAPDH MDH
atoms. E.g. G-3-P 1,3-bisPG, Malate OAA
2 Transferases Transfer functional groups between donor and acceptor
molecules. Kinases are specialized transferases that
regulate metabolism by transferring phosphate from ATP to
other molecules.
3 Hydrolases Add water across a bond, hydrolyzing it
aldolase
E.g. F-1,6-BP DHAP + G-3-P
4 Lyases Add water, ammonia or carbon dioxide across double bonds,
or remove these elements to produce double bonds.
enolase
E.g. 2-PG PEP
5 Isomerases Carry out many kinds of isomerization: L to D isomerization,
mutase reactions (shifts of chemical groups) and others
GPI

E.g. G-6-P F-6-P

6 Ligases Catalyze reactions in which two chemical groups are joined
(or ligated) with the use of energy from ATP.
PC
E.g. Pyruvate OAA
Coenzymes:
Enzymes – classified on the basis of their composition.

Enzymes composed of wholly protein are known as simple

enzymes.

 Enzymes composed of protein plus a relatively small organic

molecule are known as holoenzymes.

 The covalent binding between the apoenzyme and non-protein

components is termed a prosthetic group.

 The non-covalent binding between the apoenzyme and non-

protein components is termed a coenzyme.
 Prosthetic group

Non-protein coenzymes
component
Non-protein
component

apoenzymes

Simple
protein

 Many prosthetic groups and coenzymes are water-soluble

derivatives of vitamins.

 The followings are common coenzymes and reactions they

mediate:
Coenzymes Reactions mediated (group transfer)
(precursor/vitamins)

Biotin Carboxylation (CO2) Pyruvate OAA

Cobalamin (B12) Alkylation (methyl group), intramolecular rearrangements, and
ribonucleotide reduction
CoenzymeA Acyl transfer (R-C=O group)
Pyruvate + CoA + NAD ——> CO + acetyl-CoA + NADH + H
+ +
(panthothenate) 2

Flavin coenzymes (B2) Oxidation-reduction (hydrogen atoms) (1 or 2 e- transfer)

Lipoic acid Acyl group transfer

Nicotinamide Oxidation reaction (hydrides ions H:-, 2 e- transfers)
coenzymes (niacin)
Pyridoxal phosphate Amino group transfer ( and many other reactions)
(B6)
Tetrahydrofolate (Folic One-carbon transfer
acid)
Thiamine Aldehyde transfer
pyrophosphate (B1)
Uridine diphosphate Sugar transfer (hexose unit)
(UDP)
Industrial applications

Detergent: We use enzymes in our everyday life for example in

detergents containing proteases and lipases to remove stubborn
stains.

Fruit juices: Pectinase is added to digest pectin (the “glue” that

holds plant cells together) thus reducing wastage, enhance clarity
etc.

Meat tenderizer: Papain is used as a meat tenderizer.

 Clean up oil spills: Certain enzymes produced by microbes
can digest harmful oil and convert them into harmless carbon
dioxide and water.

 Animal feed: enzymes are added to animal feed to increase

digestibility.
Natural digestive enzymes

Natural digestive enzymes – enzymes that are naturally found in

fruits and vegetables, which affects the activity of the digestive
system by increasing absorption, utilization and assimilation of
food.

Good source of proteolytic enzymes that help in proper

breakdown of protein.

The natural digestive enzymes enhance metabolism and help in

proper digestion of food.
 It also eliminates stomach pain, bloating, abdominal cramps
and indigestion.

 It also helps to reduce weight by breaking down fat molecules.

It helps to reduce toxic effects of certain foods by eliminating
free radicals from the body.

 Some enzymes also have an anti-oxidant activity that purifies

blood.

 Natural enzymes – are equally powerful as chemical tablets or

syrups containing digestive enzymes.
Some common local medicinal plants and their uses

Papain
 Source: Carica papaya

 Common name/s:

 English: Pawpaw
 Hausa: Gwanda
 Yoruba: Ibepe
 Ibo: Okoro-beke
 Papain enzyme – plant-based digestive enzyme that comes
from the dried purified latex obtained from the unripe fruit of
Carica papaya

 Fruit is edible – rich in

 Source of proteolytic enzyme – papain

Vitamin A
Vitamin C
Chemical nature
 It is also known as "Vegetable Pepsin" because it contains
similar proteolytic enzymes of pepsin. Papain can act in acid,
neutral or alkaline media.
 The enzyme becomes active in an environment of 6.0 - 8.0 pH
and required temperatures above normal human body
temperature.

Method of preparation
The matured unripe fruits are incised on four sides to allow free
flow of latex, which solidifies quickly upon collection in aluminum
trays.
 Repeated incisions are made at weekly intervals until
exudation of latex ceases.
 The solidified latex is treated with an antioxidant like potassium
bisulphate and then sieved to remove any extraneous matter.
 Drying is done in the sun or by artificial – spray drier or in a
shelf drier at 55oC to 60oC.
 The crude papain is dissolved in water and then precipitated
with alcohol. It can also be purified by affinity chromatography
or by HPLC methods
Uses: Medicinal, Pharmaceutical and Cosmetic

 Used in treating infected wounds, sores, ulcers, chronic

diarrhea, tumors, hay fever, esophageal obstruction due to
meat impact, catarrh and psoriasis.

 Currently used as digestive aids and in preparations to control

edema and inflammation associated with surgical or accidental
trauma, infections, or allergies.
 It is also used in certain face
creams, cleansers, face lift
formulations, and dentifrices and
as an active ingredient in
enzyme cleaners for soft lenses.

 Other – modifying dough in baking, clarifying fruit juices, in fish

oil extraction, in manufacture of collagen heat-sealable films
and manufacture of sausage casings
 Chymopapain is used in the treatment of degenerative
intervertebral disc disorders (low back pain) due to herniated
lumbar disc.
 It is also used to relieve the symptoms of episiotomy.
 It is used for the tenderization of meat in meat packing
industry.
 Papain is used as a digestive aid ingredient in capsules,
tablets as it has pepsin like activity.
 Examples
Bromelain
 Scientific name:
Ananas comosus

 Common name/s:
 Hausa: Abarba
 Yoruba: Ope
oyinbo

 Ibo: Akwuolu
 Bromelain – plant-based digestive enzyme that comes from
the Ananas comosus
 Fruit is edible – rich in
 soluble mono and disaccharides, inorganic acids and
vitamins.
 Source of proteolytic enzyme – bromelain

Chemical constituent
Peptidase, ananain, Bromelain, etc.

Fruit is rich in soluble mono and disaccharides, inorganic acids

and vitamins.
Method of preparation

Extraction of bromelain from fruit and skin

Fruits are cut into small pieces, weighed, macerated and the
juice is then obtained.
The juice is then pressed and filtered through cheese cloth.

The pH of the juice is adjusted to 6.

Ammonium sulphate is then added until saturation to precipitate

the enzyme.
 Partial purification is done by re-dissolving crude enzyme in
NaCN and repeatedly precipitating it, firstly with 0.6%
ammonium sulphate and then with acetone.
 The precipitate is thoroughly washed with acetone and ether
and dried in vacuum oven at low temperature.

Extraction of bromelain from stem

Matured stem – is collected after final fruit harvest and freed from
leaves and sucker stems.
Stems is then weighed and placed in scrapping machine.
 Juice is pressed and filtered through cheese cloth.
 Two volume of acetone is initially added to two volumes of
stem juice.
 The precipitate formed is discarded as it has low enzyme
activity, poor color and stability.
 The addition of another volume of acetone is done which
precipitate the main enzyme fraction.
 This is then collected by centrifugation and washed with
acetone and diethyl ether and dried in vacuum oven.
Uses:
Used to treat inflammation and edema associated with surgical
or accidental trauma, infections or allergies.
Used in certain cosmetics such facial cleansers and bath
preparations.
Major uses are in meat tenderizing, manufacturing precooked
cereals, modifying dough (bread, wafers, pizza etc.)
Use to accelerate the tissue repair of post episiotomy.
Used in some vitamin and herbal formulations, mainly as a
digestive aid.
 Examples