MODULE 2

APPLICATION
OF
BIOMOLECULES
Dr. SHREYA KANTH
ASSISTANT PROFESSOR
DEPARTMENT OF CHEMISTRY
ACHARYA INSTITUTE OF TECHNOOGY
EMAIL: shreya2901@acharya.ac.in
APPLICATION OF
BIOMOLECULES
 Carbohydrates in cellulose-based water
filters production, PHA and PLA in
bioplastics production
 Nucleic acids in vaccines and diagnosis
 Proteins in food production
 lipids in biodiesel and detergents
production
 Enzymes in biosensors fabrication, food
processing, detergent formulation and
textile processing.
CARBOHYDRATES
They are the most abundant class of
biomolecules on earth.
 Carbohydrates, which are made up
primarily of molecules containing
atoms of carbon, hydrogen, and
oxygen, are essential energy sources
and structural components of all life.
 They are built from four types of
sugar units—monosaccharides,
disaccharides, oligosaccharides, and
polysaccharides.
Fiber is an example of a complex carbohydrate. The main role of fiber is to
keep the digestive system healthy.
 They may help prevent stomach or intestinal problems, such as
constipation. They may also help lower cholesterol and blood sugar. Fiber is
found in many foods that come from plants, including fruits, vegetables,
nuts, seeds, beans, and whole grains.
 Cellulose is another best example for carbohydrates.
 Cotton fibres are the purest form of cellulose, nature's most abundant
polymer. Nearly 90% of the cotton fibres are cellulose.
 The non-cellulosics are located on the outer layers or inside the lumens of
the fibers whereas the secondary cell wall is purely cellulose.
Cellulose, a complex carbohydrate (or polysaccharide), consists of
3,000 or more glucose units.
Abundant, easily renewable, and biodegradable.
Due to intermolecular and intramolecular hydrogen bonding
between the hydroxyl groups of the neighboring cellulose
chains, cellulose is insoluble in water, and is difficult to
dissolve with common organic solvents.
Taking benefit of these properties Cellulose-based air/water filters
have been developed.
Cellulose-based water filters are filters made from
natural material cellulose, a carbohydrate polymer found in
plants (Plant cell walls).
These filters are used to clean and purify water by trapping
and removing impurities like dirt, bacteria and other particles.
These filters are renewable, environmentally friendly and
easy to use.
These filters are used for household, industrial and
agricultural applications.
 Production of filters from
•cellulose
Water based fibres
filters have the potential to be made
affordable, lightweight and biodegradable.

• Focus on creating biobased membranes for micro- and

ultrafiltration from cellulose nanofibrils (CNFs).

• Filters based on cellulose pulp facilitate water

percolation but they do not sufficiently remove
bacteria through size exclusion; other techniques are
therefore needed to achieve a bacteria-reducing effect.

• Incorporating antibacterial metal nanoparticles into

cellulose-based water filters; both silver nanoparticles
(AgNPs) and copper nanoparticles (CuNPs) are known to
have good antibacterial effects.

• An alternative method is to use positively charged

filters that adsorb negatively charged bacteria onto
the surfaces of the filters.
SYNTHESIS OF CELLULOSE FIBERS
.
Cellulose based nanomaterials used in water
treatment
Cellulose filter papers are versatile and diverse tools for
microfiltration, that work by trapping particulates within a random
matrix of cellulose fibers. Cellulose filter papers can be categorized
as quantitative or qualitative, depending on their application.
Production of biopolymers
Biopolymers which are produced with the help of microorganisms
require specific nutrients and controlled environmental conditions.
They are produced either directly via fermentation or by chemical
polymerization of monomers, which are in turn produced through
fermentation.
Polylacticacid (PLA)
•PLA is bioplastic both biobased and biodegradable under industrial
composting conditions (at a high temperature, around 58 °C).
•Good mechanical properties, processability, renewability, and non-
toxicity.
•PLA has better durability, transparency, and mechanical strength.
Polyhydroxyalkanoate (PHA)
•PHAs are a significant polymer family that are 100% bio-based and
bio-degradable.
•PHAs are microbiologically produced polyesters that have tunable
physical and mechanical properties.
•Low environmental impact due to biodegradability and non-toxicity
nature.
•Promising candidates for sustainable future manufacturing
•Ranging from brittle thermoplastics to gummy elastomers
•PHAs' properties can be altered by the selection of bacteria,
fermentation conditions, and substrate.
•Due to their flexible properties, PHAs can substitute PP,
polyethylene (PE), and polystyrene (PS), which are the main
polymers of today's global polymer market.
 PLA and PHA
POLY LACTIC ACID [PLA] AND
comparative properties
POLYHYDROXY ALKANOATES [PHA]
• Bioplastics - generated from natural
resources such as starches and
vegetable oils.
• Bioplastics are basically classified as
bio based and/or biodegradable.
• Bioplastics are referred to as bio based
when the focus of the material is on
the origin of the carbon building block
and not by where it ends up at the end
of its cycle life.
• Bio plastics are said to be
biodegradable if they are broken down
with the effect of the right
environmental conditions and microbes
which in turn use them as a food
source.
• The bioplastics are considered
compostable, if within 180 days, a
complete microbial assimilation of the
fragmented food source takes place in
a compost environment.
STEPS IN PRODUCING BIOPLASTIC PHA AND PLA

Bacillus species
 Nucleic acids in vaccines and
• diagnosis
Composition-Deoxyribonucleic acid (DNA)
and Ribonucleic acid (RNA) are composed
of nucleotides. Nucleotide is made up of
nucleoside units (Base + Sugar)
• 5-carbon sugar, a phosphate group and a
nitrogenous base.

• Serve as the primary information-carrying

molecules

• Important role in directing protein

synthesis.
• Strings of nucleotides are bonded to form
helical backbones
Single stranded for RNA,
Double stranded for DNA
• Nucleobases adenine, cytosine, guanine,
thymine, and uracil. Structure of the main forms of Nucleic acids
• Thymine occurs only in DNA and uracil
only in RNA
DNA Vaccines for Rabies
Rabies is a preventable viral disease most often transmitted through
the bite of a rabid animal. The rabies virus infects the central
nervous system of mammals, ultimately causing disease in the brain
and death. Most rabies cases reported to the Centers for Disease
Control and Prevention (CDC) each year occur in wild animals like
bats, raccoons, skunks, and foxes, although any mammal can get
rabies.
DNA Vaccine
• A DNA vaccine, using a pCi-neo plasmid encoding the
glycoprotein gene of a Mexican isolate of rabies virus, was
developed to induce long-lasting protective immunity
against rabies virus in dogs.

• A range of parameters including physical, physiological,

clinical, immunological, hematological along with
histopathology profiles of target organs was monitored to
assess the impact of vaccination.

• There were no observational adverse effects despite high

dose administration of the DNA vaccine formulation.

• Thus, this study indicates the safety of next generation of

Steps in the production of DNA Vaccines
RNA VACCINES FOR COVID-19
• Coronavirus disease (COVID-19) is an infectious disease caused
by the SARS-CoV-2 virus.
• Messenger RNA, or mRNA technology, instructs cells to make a
protein that generates an immune response in the body, thus
producing the antibodies that provide protection against a
disease.
• It is the basis for the Pfizer/BioNTech and Moderna COVID-19
vaccines being used by governments worldwide, and in the UN-
supported COVAX global vaccine solidarity initiative.
• mRNA is a molecule that provides cells with instructions for
making proteins.
• mRNA vaccines contain the instructions for making the SARS-CoV-
2 spike protein. This protein is found on the surface of the virus
that causes COVID-19.
Steps in the production of RNA Vaccines
• The mRNA molecule is essentially a recipe, telling the cells
of the body how to make the spike protein.
• COVID-19 mRNA vaccines are given by injection, usually
into the muscle of the upper arm.
• After the protein piece is made, the cell breaks down the
instructions and gets rid of them.
• The mRNA never enters the central part (nucleus) of the
cell, which is where our DNA (genetic material) is found.
Your DNA can't be altered by mRNA vaccines.
• The cell then displays the protein piece on its surface.
• Our immune system recognizes that the protein doesn't
belong there and begins building an immune response and
making antibodies.
Proteins in food production
• Proteins are macromolecules that comprise one or more long
chains of amino acid residues.
• Proteins are found throughout the body (in muscle, bone, skin,
hair, and virtually every other body part or tissue).
• Proteins functions-
Providing structure to cells and organisms (Lipoproteins,
glycoproteins)
Storage proteins (Ferritin- stores iron)
• Applications of proteins in food production
1. Meat analogues
2. Whey proteins
 Structures of naturally
occurring 20 amino acids in
a polypetide
WHEY PROTEIN

•Whey protein offers several advantages that impact both internal

health and external appearance.
•It helps in muscle repair and strengthening, contributes to a slower
aging process and supports weight management.
PLANT BASED PROTEINS:
Plant protein is simply a meaningful food source of protein which is
from plants. This group can include pulses, tofu, soya, tempeh,
seitan, nuts, seeds, certain grains and even peas. Pulses are a large
group of plants, which include chickpeas, lentils, beans (such as
black, kidney and adzuki beans) and split peas.
Plant proteins are highly nutritious – not only as good sources of
protein, but also because they provide other nutrients such as fibre,
vitamins and minerals. Our intake of fibre tends to be too low,
however by incorporating certain plant proteins into your diet, such
as pulses, peas and nuts, you can easily boost your fibre intake.

Diagrams of production of plant-based

products
Meat analogues, can be defined as products that mimic
meat in its functionality, bearing similar appearance, texture,
and sensory attributes to meat.
Meat analogues find raising interest of many consumers
who are looking for indulgent, healthy, low environmental
impact, ethical, cost-effective, and/or new food products.
MEAT ANALOGIES
ipids in Biodiesel and detergent production
• Lipids, are a broad group of naturally-occurring molecules
which includes fats, waxes, sterols, fat-soluble vitamins
(such as vitamins A, D, E and K), monoglycerides,
diglycerides, phospholipids, and others.
• The functions of lipids include storing energy, signaling,
and acting as structural components of cell
membranes. Lipids have applications in the cosmetic and
food industries, and in nanotechnology.
BIODIESEL
 Renewable and natural domestic fuel
extracted from animal fats or Vegetable oils
(soya bean; Rapeseed; Jatropha; Palm oil).
 Biodiesel are the methyl esters obtained
from the organic feedstock.
 Biodegradable, non toxic
 Emission of green house gases is low
compared to green house
 Promotes environmental protection and
energy security.
 Generates rural employment opportunities
RAPESEED JATROPA

FATTY
ACID

ESTER

SOYABEAN PALM OIL

ESTERIFICATION

TRANSESTERIFICATION
STEPS IN
TRANSESTERIFICATION
 Treatment of raw material
 Alcohol- catalyst mixing
 Chemical reaction
 Separation of reaction
products
 Purification of reaction
products
DISADVANTAGES
 Slightly higher consumption due to the
calorific value of biodiesel
 Nitrous oxide (NOx) emissions are higher
compared to diesel fuel
 Higher freezing point than diesel fuel
 Less stable than diesel fuel, storage of
biodiesel ( more than six months) is not
recommended.
 Higher production costs, and
 Processing methods should be effective
and cost-effective
DETERGENT PRODUCTION
LIPIDS AS CLEANING
AGENTS
 Lipids as cleaning agents or detergents is
based on the dissolving of grease and oils.
 As they are composed on hydrophobic and
hydrophilic regions, which allows them to
surround around grease and oils, effectively
breaking them into smaller particles that can
be removed easily.
 This property allows lipids to be used in
cleaning agents such as soaps, shampoo,
detergents.
 The amphiphilic character of these
substances also make them strong
surfactants.
 Personal care products
 Industrial cleaning – Metal cleaning, degreasing,
stain removal
 Laundry detergents

 Properties – Emulsification (process of mixing

two immiscible phases. Eg. Oil and water) and
degreasing (to grease, oil etc. by treating with a
chemical)

 Examples: Soaps, fatty acids (stearic acids used in

soaps and shampoos)
 Glycerol
 Fatty alcohols
 Lipid-based cleaning agents is applied to the
surface, hydrophobic regions of the lipid
molecule surround and dissolve the grease and
oils , while the hydrophilic regions interact with
water , allowing the mixture to be rinsed away.

 Combination of lipid and water forms emulsion, which

helps in removing the dirt and debris

 Some lipids have additional properties, such as

foaming or lathering capabilities, that can enhance
the cleaning performances.

 Example: foaming agents in shampoos; lathering

properties in soap
ADVANTAGES

 Biodegradability
 Renewable sources
 Effectiveness
 Mildness
 Cost effective
DISADVANTAGES

 Stability
 Compatibility
 Cost
 Availability
 Performance
 Regulation
ENZYMES
The important class of biomolecules, which are
mostly proteins, that assist in speed up of
metabolism, or the chemical reactions in our
bodies. Enzymes are biological catalysts that
perform a vast majority of biochemical reactions in
living organisms.
The popular six class of enzymes are hydrolases,
oxidoreductases, lyases, transferases, ligases
and isomerases.
Enzymes in biosensors fabrication, food
processing, detergent formulation and textile
BIOSENSORS FABRICATION
Biosensor is a device that uses a living organism or biological
molecule, especially enzymes or antibodies to detect the
presence of chemicals.

Examples: Blood glucose meters, Continuous glucose

monitors, Cholesterol meters
GLUCOSE OXIDASE IN BIOSENSORS:
Glucose oxidase (GOx) is widely used enzyme in glucose biosensor
due to its better stability and relatively low cost. GOx catalyses the
redox reaction and transfer electrons from enzyme active sites to
electrode for glucose level analysis in blood samples. Amperometric
glucose biosensor was fabricated by immobilizing glucose oxidase
(GOx). Glucose oxidase (GOx), the most popular enzyme used for
glucose detection, is able to reduce oxygen to hydrogen peroxide
while at the same time transforming glucose to d-glucono-1,5-
lactone. Quantification of glucose can be achieved based on either
the detection of the hydrogen peroxide produced, or the oxygen
consumed.

Application of enzymes in Biosensors

ENZYMES IN FOOD PROCESSING
Enzymes are used in various food industries such as
•Dairy industry
•Brewing industry
•Baking industry
•Wine and juice making industry
Sources of food enzymes (plant, animal, microbial,
and recombinant).

Enzymes have been used inadvertently or deliberately in food

processing since ancient times to make a variety of food products,
such as:

 breads,
 fermented alcoholic beverages,
 fish sauces,
 cheeses.

Enzymes have been traditionally produced by

 extraction and fermentation processes from plant and animal

sources,
 from a few cultivatable microorganisms.
Sources of food enzymes (plant, animal, microbial,
and recombinant)

Industrial enzymes have traditionally been derived

from:

Plants: α-amylase, β-amylase, bromelain, β-

glucanase, ficin, papain, chymopapain, and
lipoxygenase

Animals: trypsins, pepsins, chymotrypsins,

catalase, pancreatic amylase, pancreatic lipase,
and rennin (chymosin)

Microorganisms: α-amylase, β-amylase, glucose

isomerase, pullulanase, cellulase, catalase, lactase,
pectinases, pectin lyase, invertase, raffinose,
Enzymes used in food industry have mainly microbial origin.
Advantages of microorganisms as a source for
enzyme production:

 Easy and fast grow.

 Take small space to cultivate.
 Relatively cheep culture compounds.
 Their use as enzyme source is not affected by
seasonal changes and climatic conditions and
are thus more consistent.
 Possibility for tight control of culture conditions.
Even though all classes of enzymes are expected to occur
in all or most microorganisms, in practice, the great
majority of industrial microbial enzymes are derived from
only a very few GRAS (generally recognized as safe)
species.

The predominant microorganisms used for industrial

production of enzymes for food purposes are:

 Aspergillus species,
 Bacillus species,
 Kluyveromyces species.

Limited use is due to: co-production of harmful toxins.

There is a need for stringent evaluation for safety at high

cost before they can be put to use for food production.
MARKET ENZYME FUNCTION
Dairy Rennet Coagulant in
cheese
production
Lactase Hydrolysis of
lactose to give
lactose-free
milk products
Protease Hydrolysis of
whey protein
Catalases Removal of
hydrogen
peroxide
MARKET ENZYME FUNCTION
Brewing α- Amylase hydrolyse
starch

α- glucanase Hydrolyse
glucan

Protease Increase soluble

proteins and
free amino
nitrogen
Amyloglucosida Increase
se glucose content
The Brewing Process
1. Malting: Raw barely is heated and dried with crushing
of the grain. The aim of malting is to isolate enzymes
needed for brewing. The barely grain is then passed
through grits mils to form grits.
2. Mashing:The grits mils is introduced to hot water and
left to activate the natural enzyme that help to
breakdown starch into sugar.
3. Lautering: The grits are separated from water. The
sugar-water saturate is drained off from grains the
sticky liquid is called wort.
4. Boiling: The wort is boiled for atleast 60-120 minutes.
5. Fermentation: Boiled strained and cooled wort is
transferred to fermentation vessel and yeast is added.
Yeast help to convert sugar into alcohol releasing carbon
dioxide during the process.
6.Botling and aging: After the fermentation beer need
to mature in order to develop flavour and for
carbonation effect. The beer is kept for aging from few
month to few years followed by bottling then ready for
commercial use.
MARKET ENZYME FUNCTION
Baking Maltogenic Modifies starch
amylase while most of
the starch starts
to gelatinise
Glucose oxidase Oxidizes
glucose and
produce
gluconic acid
and hydrogen
peroxide
Pentosenase Increase the
volume and
lightness and
improve the
crumb quality of
wheat breads.
MARKET ENZYME FUNCTION
Wine and fruit Pectinase Prevents pectin
juice from forming
haze; break
down pectin
and releases
methanol
β- Glucanase Accelerates all
biological
mechanisms
NZYMES IN DETERGENT FORMULATIONS

Enzyme-based detergents are fast emerging as

an alternative to synthetic detergents owing to
their
Biodegradability
Low toxicity
Non-corrosiveness
Environmental friendliness
Enhanced cleaning properties
Increased efficiency and stability in different
formulations
They are therefore also being referred to as
DIFFERENT ENZYMES USED IN DETERGENTS

Enzyme class Application

Protease Protein stain removal
Amylase Starch stain removal
Lipase Lipid stain removal
Cellulase Cleaning, colour
clarification, anti-
redeposition (cotton)
Mannanase Mannanan stain
removal (reappearing
stains)
ENZYMES IN TEXTILE PROCESSING
Textile processing in which enzymes are
widely used for:

1)Desizing: Process of removing the starch material

from warp yarns
2)Scouring/bioscouring: Process of removing all the
natural impurities like oil, wax, fat from the surface
of fabric to make it more hydrophillic and white
3)Bleaching: Purpose of cotton bleaching is to
decolourize natural pigments
4)Ageing of denim: Cellulase enzymes are used in
denim garment processing to get stone wash look
without using pumice stone
5)Biopolishing: Enzymes used to remove the fabric
pilling formed on the surface of fabric
6)Degumming of Silk: Process of removing sericin
or silk gum from silk
se of enzyme in textile industry
Enzyme Class Application

Cellulase Denim finishing, cotton

softening
Amylase De-sizing

Pectate lyase Scouring

Catalase Bleach termination

Laccase Bleaching

Peroxidase Excess dye removal

 QUIZ
1.Which of the following is
not included in the main class
of biomolecules
a)Lipids
b)Nucleic acids
c)Phosphates
d)carbohydrates
 Which of the following macromolecu
ontains carbon, nitrogen, oxygen,
hosphorous and hydrogen in the
ructure
)Carbohydrate
)Nucleic acid
Lipid
)protein
3.Sugar, starch, cellulose
and glucose are examples of
what class of
biomolecules
a)Lipids
b)Proteins
c)Carbohydrates
d)nucleic acid
4.Which among the following
is not present in DNA
a)Thymine
b)Uracil
c)Adenine
d)Cytosine
5. Which enzyme is used in
the analysis of glucose in
blood sample
a)hydrolase
b)Glucose oxidase
c)Lipase
d)amylase