MODULE 2
APPLICATION OF BIOMOLECULES
• Carbohydrates in cellulose-based water filters
production
• PHA and PLA in bioplastics production
• Proteins in food production
• Lipids in biodiesel and detergents production
• Enzymes in biosensors fabrication, food processing,
detergent formulation and textile processing.
• Nucleic acids in vaccines and diagnosis
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Carbohydrates - Applications
1) Cellulose – Based Water Filter
• Cellulose is a polymer made up of glucose subunits.
• Cellulose is among the most commonly used fibers in
filtration media.
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• Cellulose filter papers are versatile and diverse tools for
microfiltration, that work by trapping particulates within
a random matrix of cellulose fibers.
• This versatile, proven, and cost-effective material is
made from renewable resources and compatible with a
multitude of filter production processes, such as
embossing, corrugating, and pleating.
• Similar to traditional papermaking, cellulose media is
manufactured using a wet laid process.
• The result is a filtration media platform suitable for a
broad range of filtration applications, with the ability to
capture an array of particles down to nanoparticles.
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• Filters based on cellulose pulp fibers incorporating
antibacterial metal nanoparticles into cellulose-based
water filters - both silver nanoparticles (AgNPs) and
copper nanoparticles (CuNPs) are known to have good
antibacterial effects
• Cellulose filter media can be made from pure cellulose
fibers or cellulose fibers mixed with synthetic fibers or
glass fibers for enhanced properties.
• Cellulose can be prepared with different functional
groups in different sizes and derivatives including
cellulose nanocrystals (CNCs), cellulose nanofibrils
(CNFs), and TEMPO (2,2,6,6-tetramethylpiperidine-1-
oxyl radical)-mediated oxidized cellulose nanofibrils
(TOCNF).
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• These nanoscaled entries can be prepared via several
methods including mechanical, chemical, and biological
processes
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• The Four Main Nanocellulose Types
i. MFC – Microfibrillated Cellulose
• Produced mainly by mechanical treatment of cellulose fibers.
• Consists of long, entangled fibrils.
• Properties
• High surface area
• Strong mechanical strength
• Flexible network
• Applications
• Foams
• Aerogels
• Hydrogels
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ii. CNF – Cellulose Nanofibrils
• Similar to MFC but typically more refined nanoscale fibrils.
• Characteristics
• Long and flexible
• High aspect ratio (very long compared to width)
• Applications
• Membranes
• MF (microfiltration)
• UF (ultrafiltration)
• NF (nanofiltration)
• RO (reverse osmosis)
• These are used for water purification and filtration.
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iii. CNC – Cellulose Nanocrystals
• Produced by acid hydrolysis of cellulose.
• Removes amorphous regions and leaves rigid crystalline rods.
• Properties
• Very stiff
• High crystallinity
• High strength
• Applications
• Paper reinforcement
• Thin films
• Nanocomposites
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iv. TOCNF – TEMPO-Oxidized Cellulose Nanofibrils
• These are chemically modified CNF.
• Produced using TEMPO oxidation.
• Adds carboxyl groups (–COO⁻) to the surface.
• Advantages
• Better dispersion in water
• Easier functionalization
• Higher surface charge
• Applications
• Fibers
• Electrospinning
• 3D printing materials
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• Cellulose filtration media offers a host of benefits,
including:
• Renewable fiber sources
• High strength and durability
• Large surface area
• Chemical stability
• Temperature stability
• Strong adhesion to glass
• Good value
• Cellulose can lose mechanical strength in certain
chemical or high-temperature environments and absorb
water in high-humidity conditions, potentially altering its
filtration properties
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2) Bioplastics
• Bioplastics are one type of plastic which can be
generated from natural resources
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• Bioplastics are basically classified as bio based and/or
biodegradable.
• Not all bio-based plastics are biodegradable and
similarly not all biodegradable plastics are bio based.
• Bioplastics are referred to as bio based when the focus
of the material is on the origin of the carbon building
block and not where it ends up at the end of its cycle
life.
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Production of Biopolymers
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• PHAs can be defined as a family of intracellular
biopolymers that are synthesized via various bacteria as
intracellular carbon and energy storage granules.
• These renewable polyesters can be produced by various
microorganisms in response to various stress conditions
(for example, excess carbon or limited phosphate,
nitrogen, sulfur, or oxygen) and provide protection from
nutrient starvation and extreme conditions.
• PHAs characteristics include, water insolubility, relative
resistant to hydrolytic degradation, biocompatibility and
suitability for medical applications, as well as nontoxicity.
• Although, PHAs are not water soluble, they are still
degradable and biocompatible.
• In addition, PHAs are considered less sticky than other
polymers once heated, and they sink in water which
facilitates their anaerobic biodegradation in sediments.
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• PHAs are both compostable and biodegradable in marine
environments.
• Due to their biodegradable nature, PHAs are intended to
replace synthetic non-degradable polymers for various
applications, such as: packaging, fast food, medicine,
biomedical, and agricultural applications
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PLA- (Polylactic acid) •
• Polylactic acid or polylactide (PLA) is a polyester
derived from renewable biomass, typically from
fermented plant starch, such as corn, cassava,
sugarcane or sugar beet pulp.
• PLA is a polyester (polymer containing the ester group)
made with two possible monomers or building blocks:
lactic acid, and lactide.
• PLA is biodegradable under appropriate conditions and
is generally regarded as food safe since it decomposes
back into its lactic acid building blocks, which are non-
toxic.
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• However, it must be noted that PLA does not degrade
naturally in the environment and must be composted
under a special set of conditions such as a temperature
of at least 60 °C, and humidity of 90 % which must be
maintained over a period of 60 to 90 days.
• PLA decomposes into carbon dioxide, lactic acid, and
water.
• PLA is used in food packaging and disposable cutlery
and can be formed into fibers for clothing.
• It is also one of the most widely used 3D printing
filaments for fused deposition modeling (FDM) due to
its low melt temperature and its ease of use.
• PLA is compostable but may stay for up to a thousand
years in the marine environment
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PROTEINS AS FOOD
• All biologically produced proteins can be used as food.
• However, proteins used as foods can be defined as
those that are easily digestible, nontoxic, nutritionally
adequate, functionally useable in food products and
available in abundance.
• Dietary proteins are found in
• animal based foods
• plant-based foods
• alternative sources such as algae, bacteria, and fungi
(mycoproteins)
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Animal Based Proteins
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Plant Based Proteins
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• 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.
• Consumer demand for plant protein-based products is
high and expected to grow considerably in the next
decade.
• Factors contributing to the rise in popularity of plant
proteins include:
• potential health benefits associated with increased intake of
plant-based diets
• consumer concerns regarding adverse health effects of
consuming diets high in animal protein (e.g., increased
saturated fat)
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• Increased consumer recognition of the need to improve the
environmental sustainability of food production
• Ethical issues regarding the treatment of animals; and
• General consumer view of protein as a “positive” nutrient (more
is better).
• While there are health and physical function benefits of
diets higher in plant-based protein, the nutritional
quality of plant proteins may be inferior in some
respects relative to animal proteins
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1) Whey protein
• Whey protein is a mixture of proteins isolated from
whey, the liquid material created as a byproduct of
cheese production.
• The proteins consist of α-lactalbumin, β-lactoglobulin,
serum albumin and immunoglobulins.
• Glycomacropeptide also makes up the third largest
component but is not a protein.
• The primary usage of whey protein supplements is for
muscle growth and development.
• Eating whey protein supplements before exercise will
will enhance the body's protein recovery and synthesis
after exercise because it increases the free amino acids
in the body's free amino acid pool.
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• It is the largest by product of the dairy industry both in
terms of volume and milk solids.
• Through new technologies, whey and its components
become versatile ingredients and have high economic
values.
• Whey and whey components are viewed as value-
added ingredients in infant formulae, sports nutrition
foods and beverages, and other food products.
• Whey products improve the texture, extends shelf life,
provides stability, improves flow properties, enhances
both the taste and the colour thus can be used as a
food ingredient.
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2) Meat analogues
• Analogue can be defined as compound that is
structurally similar to another but differs slightly in
composition
• Meat analogues, can be defined as products that mimic
meat in its functionality, bearing similar appearance,
texture, and sensory attributes to meat.
• Meat analogues are plant-based products designed to
replace meat.
• Production of meat analogues has been on the increase,
targeted at satisfying consumers’ desire for indulgent,
healthy, low environmental impact, and ethical meat
substitutes
• Meat analogs are formulated from soy protein, wheat
gluten, mycoproteins, and proteins from other legumes,
cereals, nuts, mushrooms, and some vegetables
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• Compared with meat products, plant-based meat
analogues are more nutritious and healthier.
• They can be used to effectively alleviate the shortage of
meat resources.
• Although certain technologies for the production of
meat analogues still need to be developed, the
marketing prospects for plant-based meat analogues
are very high.
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Lipids - Applications
1) Biodiesel
• Biodiesel is an alternative fuel similar to conventional or
‘fossil’ diesel.
• Biodiesel can be produced from straight vegetable oil,
animal oil/fats and waste cooking oil.
• The largest possible source of suitable oil comes from
oil crops such as rapeseed, palm or soybean.
• Most biodiesel produced at present is produced from
waste vegetable oil sourced from restaurants, chip
shops, industrial food producers etc.
• The process used to convert these oils to Biodiesel is
called transesterification
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• The triglycerides are reacted with a suitable alcohol
(Methyl, Ethyl, or others) in the presence of a catalyst
under a controlled temperature for a given length of
time.
• The final products are Alkyl esters and Glycerin.
• The Alkyl esters, having favorable properties as fuels
for use in compression ignition engines, are the main
product and the Glycerin, is a by-product.
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• Biodiesel has many environmentally beneficial
properties.
• The main benefit of biodiesel is that it can be described
as ‘carbon neutral’.
• This means that the fuel produces no net output of
carbon in the form of CO2.
• This effect occurs because when the oil crop grows it
absorbs the same amount of CO2 as is released when
the fuel is combusted
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2) Cleaning Agents / Detergents
• Detergent is a surfactant or a mixture of surfactants
with cleansing properties when in dilute solutions.
• However, conventionally, detergent is used to mean
synthetic cleaning compounds as opposed to soap (a
salt of the natural fatty acid), even though soap is also
a detergent in the true sense.
• Most detergents are akyl benzenesulfonates.
• Detergents tend to be more soluble in water than soap
because the sulfonate of detergent doesn't bind
calcium and other ions in hard water as easily as the
carboxylate in soap does.
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Types of Lipids Used in Bio-Detergents
• Common lipids used in bio-detergents include
• Fatty acids
• Glycerides
• Phospholipids
• Fatty acids act as surfactants, reducing surface tension
and increasing solubility of dirt and stains.
• Glycerides help stabilize detergent formulation and
improve its overall performance.
• Lipids are
• effective in removing oil and grease stains due to their
affinity for fat
• Gentle on skin and not cause irritation
• Biodegradable reducing environmental impact of detergent
use.
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Lipid based Surfactants in Bio Detergent
• These are amphiphilic molecules that disperse soils in
water, can emulsify and solubilize dirt and stains.
• Lipid based surfactants are less harsh than traditional
synthetic surfactants, making these a preferred choice
for bio-detergents
• Lipid emulsifiers help stabilize the detergent formulation
and prevent separation of ingredients.
• These improve the
• Dispersion of active cleaning agents, ensuring uniform
cleaning performance
• Overall stability and shelf life of bio-detergents.
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Enzymes - Applications
1) Biosensors
• A biosensor is a device that uses a living organism or
biological molecules, especially enzymes or antibodies,
to detect the presence of chemicals.
• Glucose sensors are biosensors designed to detect
glucose levels, which is vital to managing diabetes and
use Glucose oxidase, a subset of oxidoreductase,
enzyme for detection.
• Glucose oxidase, along with catalase, is used in
glucose testing kits (especially in biosensors) to detect
and measure the presence of glucose in industrial and
biological solutions (e.g., blood and urine specimens).
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• The biosensor has a bienzymatic sensor phase
consisting of glucose oxidase and horseradish
peroxidase (HRP), with ferrocyanide as an electron-
transfer mediator.
• The enzyme glucose oxidase reacts with glucose and
oxygen to form gluconic acid and hydrogen peroxide.
• The hydrogen peroxide can then be used to oxidize a
chromogen or the consumption of oxygen measured to
estimate the amount of glucose present.
• Usually HRP catalyses the oxidation of ferrocyanide to
ferricyanide, consuming the hydrogen peroxide
previously generated.
• The analytical signal is the current intensity due to the
electrochemical reduction of the enzymatically
generated ferricyanide and Glucose level is estimated
by measuring the amount of ferricyanide produced
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2) Food Processing
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3) Enzymes in bio-detergents formulation and textile
processing
• Lipases break down lipid based stains like oils and fats
• Enhance the cleaning power by targeting specific types
of stains
• Biodegradable and environmentally friendly, aligning
with the principles of green chemistry
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• Enzymes such as, amylases, cellulases, pectinases,
catalase, laccase, etc. plays a major role in improving
the texture, quality, strength, and market values of the
cloth.
• For instance, amylases can be used in removing the
starch applied and coated on the yarn surface to
protect the fabrics from harsh chemical and mechanical
processing, in desizing step
• Pectinases can be used in bio-scouring step to remove
the noncellulosic contaminants
• Cellulase can be used for improving the fabric softness
and water absorbance
• Laccase and catalase can be used to improve the
whiteness in the bleaching of fabrics
• Catalase and peroxidase can be used for removal of
excess oxidants and degradation of synthetic dyes, etc.
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4) Bio-bleaching
• Biobleaching is the process of bleaching pulps with
enzymes or lignolytic fungi, thereby reducing the quantity
of the chemical bleach required.
• The use of bacteria or enzymes or biological agents in
the removal of color is termed as bio bleaching.
• General concern about the environmental impact of
chlorine bleaching effluents has led to a trend towards
totally chlorine free bleaching methods.
• Organochlorine compounds have been the matter of
concern in the pulp and paper industry.
• These compounds are produced mainly by the reactions
between residual lignin present in wood fibers and the
chlorine used for bleaching.
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• Some of these compounds are found to be toxic,
mutagenic, persistent, bioaccumulating and to cause
harm to biological systems.
• Considerable interest has been focused on the use of
biotechnology in pulp bleaching, as large number of
microbes and the enzymes produced by them are
known to be capable of preferential degradation of
native lignin and complete degradation of wood.
• Ligninolytic enzymes are most effective because it
directly attacks on lignin and play a key role in
degradation and detoxification of lignocellulosic waste
in environment
• Lignin-oxidizing species are called white rot fungi,
because they typically turn wood white as it decays.
• The major ligninolytic enzymes are laccase, lignin
peroxidase, manganese peroxidase, and versatile
peroxidase.
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Nucleic Acids - Applications
Vaccines
• Vaccines are biological preparations that train your
immune system to recognize and fight specific diseases—
without actually causing the illness.
• When a vaccine enters the body, it contains a harmless
part or weakened form of a germ (like a virus or bacteria).
This helps the body:
• Recognize the invader (called an antigen)
• Produce antibodies to fight it
• Build memory cells so it can respond faster in the future
• Vaccines are important to
• Prevent serious diseases like measles, polio, and COVID-19
• Reduce spread of infections (community protection or herd
immunity)
• Save millions of lives every year
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Types of vaccines
1) Live-Attenuated Vaccines: These vaccines use a weakened
(attenuated) form of the virus or bacteria that causes the disease,
allowing the body to develop immunity without causing the
illness.
2) Inactivated Vaccines: These vaccines use a killed version of the
virus or bacteria, providing immunity without the risk of infection.
3) Subunit, Recombinant, Polysaccharide, and Conjugate Vaccines:
These vaccines use specific parts of the virus or bacteria, such as
proteins, sugars, or capsid, to stimulate the immune system.
4) Toxoid Vaccines: These vaccines use a toxin (harmful product)
made by the bacteria, but in a weakened form, to stimulate
immunity against the toxin rather than the bacteria itself.
5) mRNA Vaccines: These vaccines use messenger RNA (mRNA) to
instruct the body's cells to produce a specific protein from the
virus or bacteria, triggering an immune response.
6) Viral Vector Vaccines: These vaccines use a harmless virus
(vector) to deliver genetic material that instructs the body's cells
to produce a protein from the virus or bacteria, triggering an
immune response.
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1) DNA Vaccine For Rabies:
• DNA vaccines, which are often referred to as the third-
generation vaccines, use engineered DNA to induce an
immunologic response in the host against bacteria,
parasites, viruses, and potentially cancer.
• Like any other type of vaccine, DNA vaccines induce an
adaptive immune response.
• The basic working principle behind any DNA vaccine
involves the use of a DNA plasmid that encodes for a
protein that originated from the pathogen in which the
vaccine will be targeted.
• Plasmid DNA (pDNA) is inexpensive, stable, and
relatively safe, thereby allowing this nonviral platform to
be considered an excellent option for gene delivery.
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• Some of the different virus vectors that have been used
to source pDNA include onco-retroviruses, lentiviruses,
adenoviruses, adeno-associated viruses, and Herpes
simplex-1
• 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.
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• Rabies is a simple, negative-stranded RNA virus that
encodes five structural proteins, that is, the
nucleoprotein (NP), the glycoprotein (G), the
phosphoprotein (P), the matrix protein (M), and the
polymerase (L).
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• DNA vaccination based on optimized formulation of
lysosome-targeted glycoprotein of the rabies virus
provides potential platform for preventing and controlling
rabies.
• DNA vaccination has been proposed as a cheaper and
efficient strategy for rabies prophylaxis, and its feasibility
has been demonstrated in a number of animal models
including companion animals, since 1994.
• DNA is typically delivered using plasmids into the
nucleus of host cells and requires crossing the nuclear
membrane, which can limit efficiency.
• Often administered with techniques like electroporation
(applying a brief electrical pulse) to enhance uptake.
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2) RNA vaccines for Covid19
• Scientists have developed a new type of vaccine that
uses a molecule called messenger RNA (mRNA),
necessary for protein production, rather than part of an
actual bacteria or virus.
• Once cells finish making a protein, they quickly break
down the mRNA.
• mRNA from vaccines never enters the central part
(nucleus) of the cell, where our DNA (genetic material)
is found and does not alter DNA.
• mRNA vaccines work by introducing a piece of mRNA
that corresponds to a viral protein, usually a small piece
of a protein found on the virus’s outer membrane.
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• 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.
• 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 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
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3) Forensics – DNA fingerprinting
• DNA fingerprinting, also called DNA typing, DNA
profiling, genetic fingerprinting, genotyping, or identity
testing, in genetics, is a method of isolating and
identifying variable elements within the base-pair
sequence of DNA
• Alec Jeffreys, a geneticist at the University of Leicester
in Britain, invented the first usable version of DNA
fingerprinting in 1984.
• Few years later, a chemical company, Imperial
Chemical Industries (ICI), launched the first kit
commercially available.
• The DNA testing process is comprised of four main
steps, including extraction, quantitation, amplification,
and capillary electrophoresis.
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• Forensic genetic fingerprinting can be defined as the
comparison of the DNA in a person’s nucleated cells
with that identified in biological matter found at the
scene of a crime or with the DNA of another person for
the purpose of identification or exclusion.
• The application of these techniques introduces new
factual evidence to criminal investigations and court
cases.
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