Module - 2

Application of biomolecules

Biomolecules play crucial roles in various aspects of life, from sustaining living organisms to
advancing technology and medicine. Here are some of the applications of various biomolecules
in various fields.

Carbohydrates

o Carbohydrates are defined as polyhydroxy aldehydes or ketones.

o Energy Source: Carbohydrates, composed of carbon, hydrogen, and oxygen, serve as
primary energy sources for living organisms. They fuel metabolic processes and provide
energy for growth and physical activities.
o Structural Components: Carbohydrates contribute to the structure of cell walls, connective
tissues, and other biological structures.
o Applications:

▪ In agriculture, carbohydrates are used as energy reserves in seeds and tubers.

▪ Water purification: Polysaccharides (Cellulose, starch) based filters have been widely
used in water purification.
▪ Starch-based bioplastics are eco-friendly alternatives to petroleum-based plastics.

Lipids:

o Energy Storage: Lipids store energy in the form of fats and oils.
o Cell Membranes: Phospholipids form cell membranes, maintaining cell integrity.
o Applications:

▪ Lipids are used in cosmetic products, such as moisturizers and lip balms.
▪ Biodiesel, derived from plant oils, serves as an alternative fuel source.

Nucleic Acids (DNA and RNA):

o Genetic Code: DNA stores an organism’s genetic information, determining

traits and functions.
o Gene Expression: RNA plays a role in protein synthesis.
o Applications:
▪ Genetic engineering uses DNA manipulation to create genetically
modified organisms (GMOs).
▪ PCR (polymerase chain reaction) amplifies DNA segments for
diagnostics and research

Proteins:

o Enzymes: Proteins act as enzymes, catalyzing biochemical reactions.

o Structural Elements: Proteins form the building blocks of cells and tissues.
 o Applications:
▪ Enzymes in laundry detergents break down stains.
▪ Antibodies are used in medical diagnostics and research.
▪ Biopharmaceuticals (protein-based drugs) treat diseases

Medical Applications:

o Drug Delivery: Biomolecules are used in drug delivery systems, ensuring

targeted and controlled release.
o Diagnosis: Biomarkers (specific biomolecules) aid in disease diagnosis.
o Scaffolds and Wound Dressing: Biomaterials based on proteins and
carbohydrates promote tissue regeneration.
o Herbal Extraction and Medicines: Natural biomolecules from plants have
therapeutic properties

Agriculture:

o Biopesticides: Biomolecules act as biopesticides (herbicides, fungicides,

insecticides) to protect crops.
o Plant Growth Promoters: Biomolecules promote plant growth and stress
tolerance.
o Nutrient Solubilization: Biomolecules enhance nutrient availability in soil

Carbohydrates in cellulose-based water filters production

Cellulose-derived water filtration systems utilize cellulose, a polysaccharide present in plant

cell walls, to eliminate impurities and pollutants from water, offering a sustainable substitute
to conventional synthetic polymer filtration methods. The exceptional mechanical robustness
and hydrophilic nature of cellulose render it an optimal candidate for water purification
applications. These filters efficiently eradicate particulate matter, microbial pathogens, and
various contaminants, enhancing water quality and potability. Embraced across both
industrialized and developing nations, cellulose-based filtration systems serve diverse
household, industrial, and agricultural needs. Moreover, they represent an eco-conscious
choice, being biodegradable and derived from renewable sources.

Properties of cellulose based water filter

Cellulose-based water filters have several properties that make them an attractive choice for
water filtration:

✓ High Porosity: Cellulose-based water filters possess a structure with high porosity,
enabling efficient removal of impurities and contaminants from water.
✓ Biodegradability: Cellulose-based water filters utilize cellulose, a biodegradable material,
minimizing environmental impact compared to synthetic polymer filters.
✓ Cost-effectiveness: Cellulose-based water filters are typically more economical than
traditional synthetic polymer filters, ensuring accessibility to a broader consumer base and
communities.
✓ Renewable Resource: Cellulose-based water filters utilize cellulose from a renewable
source, reducing reliance on non-renewable resources.
✓ Good Mechanical Strength: Cellulose-based water filters exhibit robust mechanical
strength, ensuring structural integrity and sustained performance over time.
✓ Chemical Resistance: Cellulose-based water filters demonstrate resistance to various
chemicals, including acids and bases, making them suitable for diverse water treatment
applications.
✓ Large Surface Area: Cellulose-based water filters feature a substantial surface area,
enhancing filtration efficiency and prolonging the interval between filter replacements.

Importance of cellulose based water filters

Cellulose-based water filters play a crucial role for multiple reasons:

•Ensuring Safe and Clean Water: These filters efficiently eliminate impurities and
contaminants from water, enhancing its safety and potability for household, industrial, and
agricultural purposes.

Promoting Sustainability: Derived from cellulose, a renewable resource, and biodegradable,

these filters lessen their environmental impact, fostering sustainability in water treatment.

•Enhancing Affordability: Often more cost-effective than conventional synthetic polymer

filters, cellulose-based filters broaden accessibility to a broader spectrum of users and
communities, particularly in developing regions.

•Demonstrating Versatility: Cellulose-based water filters are adaptable to various filtration

systems and can be tailored in diverse sizes and shapes to suit specific requirements.

•Serving as Eco-friendly Alternatives: Offering a green alternative to synthetic filters,

cellulose-based filters diminish reliance on non-renewable resources and mitigate waste,
contributing to environmental conservation efforts.

Construction of cellulose-based water filters

The construction process of cellulose-based water filters involves the following steps:

1. Cellulose Material Selection: Choosing the appropriate type of cellulose material based on
desired characteristics like strength, porosity, and chemical resistance. Common options
include paper, cotton, and wood fibers.

2. Cellulose Preparation: Preparing the cellulose material by cutting it into small pieces,
washing away impurities, and drying it for use.

3. Cellulose Layer Formation: Shaping the cellulose material into a layer either by stacking
or compressing it with heat and pressure.
4. Attachment to Filter Medium: Attaching the cellulose layer to a filter medium such as a
mesh or support structure to enhance stability and increase the filter's surface area.

5. Chemical Treatment: Optionally treating the cellulose layer with chemicals to alter its
properties, like enhancing hydrophilicity or incorporating antimicrobial agents.

6. Housing Assembly: The filter medium is assembled into a housing, which serves as a
structure to connect it to a water source and gather the purified water.

7. Filter Testing: The finished filter undergoes testing to confirm it meets specific criteria,
such as filtration effectiveness and flow rate.

Figure Construction of cellulose based water filter

Cellulose material used in house hold water filters

The cellulose material frequently employed in household water filters is cellulose acetate, a
synthetic derivative of cellulose. Cellulose acetate exhibits favorable traits such as effective
chemical resistance, significant porosity, and swift flow rates, which render it apt for household
water filtration. Moreover, its affordability ensures its suitability for household usage. While
paper, cotton, and wood fibers are alternative cellulose materials that can be utilized, cellulose
acetate is the preferred choice in most cases for water filtration due to its advantageous
properties.
 Figure : Cellulose acetate material

Advantages of cellulose based water filters

Cellulose-based water filters have several advantages that make them an attractive option for
water filtration:

• Environmentally friendly: Cellulose-based water filters are made from a renewable

resource, cellulose, and are biodegradable, reducing their impact on the environment
compared to synthetic polymer filters.

• Cost-effective: Cellulose-based water filters are often more affordable than traditional
synthetic polymer filters, making them accessible to a wider range of consumers and
communities.

• High porosity: Cellulose-based water filters have a high porosity structure, which allows
them to efficiently remove impurities and contaminants from water. Versatile: Cellulose-
based water filters can be used in various types of filtration systems and can be produced in
different sizes and shapes to fit specific needs.

• Good mechanical strength: Cellulose-based water filters have good mechanical strength,
allowing them to maintain their structure and perform effectively over time.

• Chemical resistance: Cellulose-based water filters are resistant to most chemicals, including
acids and bases, and can be used in a wide range of water treatment applications.

• Large surface area: Cellulose-based water filters have a large surface area, which enhances
their filtration capabilities and reduces the frequency of filter replacement.

Limitations of cellulose based water filters

Cellulose-based water filters come with certain limitations that should be considered in
selecting a filtration method:
• Limited Thermal Resistance: These filters are prone to structural degradation under high
temperatures.

• Selective Filtration Efficiency: They may not effectively remove certain contaminants like
heavy metals from water.

• Short Lifespan: Cellulose-based filters need more frequent replacement compared to

synthetic polymer filters. • Sterilization Challenge: Sterilizing cellulose-based filters
effectively can be challenging, increasing contamination risks.

• Prone to Clogging: They can easily become clogged when exposed to high contaminant
levels, reducing filtration effectiveness and requiring frequent replacement.

• Potential Taste Impact: Cellulose-based filters may alter water taste by absorbing or
releasing specific chemicals or minerals, affecting purified water quality

PHA and PLA in bioplastics production

Polyhydroxyalkanoates (PHAs) represent a category of biodegradable and biocompatible

polyesters synthesized by microorganisms, including bacteria and fungi. These compounds
serve as a type of bioplastic, deriving from renewable sources such as sugar and cornstarch.
PHAs are esteemed as eco-friendly substitutes for conventional petroleum-based plastics.

1. Polyhydroxyalkanoates (PHAs):
o PHAs are a group of biodegradable polymers synthesized by various microorganisms.
o They accumulate as intracellular granules and serve as energy storage.
o PHAs exhibit diverse properties, including flexibility, toughness, and biocompatibility.
o Their production involves microbial fermentation using renewable carbon sources (e.g.,
plant-based sugars).
o PHAs can replace petroleum-based plastics in applications such as packaging,
agriculture, and medical devices.
 Figure : Structure of PHA with different R groups

Properties of PHA

PHAs have several properties that make them ideal for use as bioplastics, including:

• Biodegradability: PHAs naturally decompose into water and carbon dioxide, lessening
their environmental impact.
• Biocompatibility: PHAs are safe for medical applications like sutures and implants, as
they don't cause adverse reactions in the body.
• Mechanical properties: PHAs possess similar strength and characteristics to
traditional petroleum-based plastics, making them suitable for diverse uses.
• Processing: PHAs can undergo shaping using standard plastic processing methods such
as injection molding, blow molding, and extrusion.

Engineering applications of PHA bioplastic

• Packaging: PHA is employed in different packaging formats like food containers,

beverage cups, and clamshell containers.
• Medical Devices: PHA's biocompatibility makes it suitable for manufacturing medical
devices such as sutures, implants, and drug delivery systems.
• Textiles: PHA is utilized in creating biodegradable textiles and composites for
construction and furniture.
• Agricultural Mulch Films: PHA is used in making biodegradable mulch films for
agriculture, aiding in soil erosion prevention and moisture conservation.
• Consumer Goods: PHA finds application in manufacturing various consumer goods
like toys, phone cases, and water bottles.
• Automotive Parts: PHA is utilized in crafting biodegradable automotive components
like air ducts and headlamp covers.
• Electronic Devices: PHA is utilized to create biodegradable components in electronic
devices like smartphones and laptops.
• Aerospace: PHA finds application in manufacturing biodegradable parts for aerospace
purposes, including insulation and cable management.
• Sporting Goods: PHA is employed in crafting biodegradable sporting equipment such
as golf tees and fishing lures.
• Construction: PHA is utilized in producing biodegradable insulation and
soundproofing materials for construction projects.

2. Polylactic Acid (PLA):

o PLA is derived from renewable resources, primarily corn starch or sugarcane.
o It is a biodegradable and compostable polymer.
o PLA has gained popularity due to its mechanical properties, transparency, and
ease of processing.
o Applications include disposable cutlery, food containers, and 3D printing
filaments.
o PLA is an excellent alternative to traditional plastics, especially for single-use
items.

Figure: Structure of Polylactic acid

Polylactic Acid (PLA) is a type of biodegradable plastic derived from natural sources like
corn starch or sugarcane. It's commonly used in packaging, disposable tableware, and 3D
printing as a sustainable substitute for traditional petroleum-based plastics. However, it's
crucial to understand that although PLA can decompose in industrial composting
settings, it might degrade slower in natural environments, potentially leading to harmful
effects on wildlife and ecosystems if not appropriately discarded.

Properties of PLA
 • Biodegradable: PLA can be broken down by tiny organisms in specialized
composting facilities, reducing landfill waste.
• Renewable: PLA comes from sustainable sources like corn starch or sugarcane,
lessening reliance on finite petroleum reserves.
• Clear/Transparent: PLA has a see-through appearance, suitable for packaging
purposes.
• Heat-resistant: While PLA melts at a relatively low temperature and isn't ideal for
high heat, it remains stable up to 60°C.
• Biocompatible: PLA is safe for use in food packaging and medical devices, being
non-toxic and compatible with living tissues.
• Stiffness and Strength: PLA exhibits decent stiffness and strength, though not as
robust as traditional petroleum-based plastics.
• Printability: PLA is popular in 3D printing due to its ease of use and good printing
capabilities.

Engineering applications of PLA

• Automotive Parts: PLA is employed in making biodegradable components for

vehicles like air ducts and headlamp covers.
• Electronic Devices: PLA is utilized in crafting biodegradable parts for electronic
gadgets such as smartphones and laptops.
• Aerospace: PLA finds application in manufacturing biodegradable parts for
aerospace purposes, including insulation and cable management.
• Sporting Goods: PLA is used in producing biodegradable sporting equipment like
golf tees and fishing lures.
• Construction: PLA is utilized in creating biodegradable insulation and soundproofing
materials for construction projects.
• Agricultural Equipment: PLA is employed in making biodegradable parts for
agricultural machinery such as seed trays and greenhouse film.
• Medical Equipment: PLA is utilized in manufacturing biodegradable components
for medical equipment like diagnostic tools and hospital beds.

Nucleic acids in vaccines and diagnosis

• DNA: DNA is the molecule responsible for carrying genetic instructions for the
development, function, and reproduction of living organisms. It's a double-helix
structure made of nucleotides containing deoxyribose sugar, a phosphate group, and
one of four bases: adenine, guanine, cytosine, or thymine.
• RNA: RNA assists in expressing genetic information stored in DNA by transferring
the message from DNA to the ribosome, where proteins are synthesized. It's a single-
stranded molecule composed of nucleotides containing ribose sugar, a phosphate
group, and one of four bases: adenine, guanine, cytosine, or uracil.
 Figure: Structure of DNA and RNA

The role of nucleic acids in both vaccines and diagnostic applications:

1. Nucleic Acid Vaccines:

o Definition: Nucleic acid vaccines use genetic material from a disease-causing
virus or bacterium (a pathogen) to stimulate an immune response against it.
o Types: These vaccines can be based on either DNA or RNA.

DNA Vaccine

•A DNA vaccine uses a fragment of viral or bacterial DNA to prompt the immune
system to respond against the pathogen.
• Mechanism: The vaccine introduces the pathogen's DNA into the body, where cells
absorb it and produce viral or bacterial proteins.
• Immune Response: These proteins are presented on cell surfaces, stimulating the
immune system to produce antibodies against the pathogen.
• Research Areas: DNA vaccines are actively studied for various diseases like cancer,
rabies, influenza, and HIV.
• Potential: Although still early in development, DNA vaccines have the potential to
transform vaccine creation and offer new treatments for different illnesses.
 Figure: Mechanism of action of DNA vaccine

DNA Vaccine for Rabies:

A rabies DNA vaccine uses a segment of rabies virus DNA to prompt the immune
system to react against the virus. Mechanism: The vaccine introduces rabies virus
DNA into the body, where cells absorb it and manufacture viral proteins. Immune
Response: These viral proteins are presented on cell surfaces, prompting the immune
system to generate antibodies against the rabies virus.
 Figure: Production of DNA vaccine

Importance of DNA vaccine for rabies

1. Effective Immune Response: DNA vaccines prompt a strong and lasting immune
response, requiring fewer doses for protection against rabies.
2. Long-term Protection: They provide extended defense against rabies due to their
ability to stimulate robust immunity.
3. Easy Administration: DNA vaccines are simple to administer, either through
injection or orally, which is beneficial in areas with limited medical access.
4. Cost-effectiveness: They're relatively inexpensive to produce compared to
traditional vaccines, enhancing accessibility in resource-limited regions.
5. Reduced Side Effects: DNA vaccines carry lower risk of side effects since they
don't contain live virus particles, ensuring safer immunization against rabies.

RNA Vaccines

• RNA vaccines utilize genetic material from pathogens, in the form of RNA, to prompt
an immune response against the disease.
• The vaccine functions by delivering the pathogen's RNA into the body, where cells
take it up and use it to generate viral or bacterial proteins.
• These proteins are then showcased on cell surfaces, triggering an immune response
and the production of antibodies targeting the pathogen.
• RNA vaccines offer advantages over traditional vaccines, such as quicker production
time and the ability to target multiple antigens.
• They can be manufactured rapidly, making them suitable for emergencies requiring
swift mass vaccination.
• RNA vaccines are deemed safer than traditional ones, lacking live virus or bacteria
that could induce illness.
• Currently, RNA vaccines are undergoing development and testing for various diseases
like COVID-19, influenza, and cancer.

Figure: Mechanism of action of RNA vaccine

Figure: Pipeline for the production of mRNA vaccine

RNA Vaccines for Covid19

• RNA vaccines for COVID-19 utilize genetic material from the SARS-CoV-2 virus in
the form of RNA to prompt an immune response against the virus.
• The vaccine introduces the virus's RNA into the body, where cells uptake it to produce
viral proteins.
• These proteins are then exhibited on cell surfaces, initiating an immune response and
antibody production against the virus.
• The Pfizer-BioNTech vaccine, the first RNA vaccine for COVID-19, received
emergency use authorization in December 2020 and has been widely administered.
• Another RNA vaccine, the Moderna vaccine, also received emergency use
authorization in December 2020.
 Figure: Mechanism of action of mRNA vaccine

Importance of RNA vaccine for Covid 19

• RNA vaccines are crucial in combating COVID-19 due to their numerous advantages
over traditional vaccines.
• They exhibit high efficacy, with Pfizer-BioNTech and Moderna mRNA vaccines
boasting approximately 95% effectiveness in clinical trials.
• Rapid development and manufacturing characterize RNA vaccines, exemplified by
the Pfizer-BioNTech vaccine's creation in under a year and quick authorization within
11 months.
• RNA vaccines offer easy modification to target new virus strains or variants, ensuring
adaptability against emerging threats.
• Considered safe, RNA vaccines prompt cells to produce harmless virus components,
like the spike protein, eliciting an immune response without live virus particles.
• RNA vaccines hold promise beyond COVID-19, potentially addressing other
infectious diseases like influenza, HIV, Zika, and even contributing to cancer
treatment.
Nucleic acids in diagnosis

DNA fingerprinting, also referred to as DNA profiling or genetic fingerprinting, is a method

employed in forensic science to distinguish individuals based on their distinct DNA profile.
This technique entails examining particular sections of an individual's DNA, known as
markers, which exhibit variation among different people.

Method of DNA fingerprinting

• SAMPLE COLLECTION: DNA is taken from a biological sample like blood, semen,
or hair. The sample is cleaned and treated to get the DNA.
• DNA AMPLIFICATION: The collected DNA is then increased using a method called
polymerase chain reaction (PCR). PCR makes many copies of a specific DNA part,
aiding in more accurate examination.
• DNA ANALYSIS: The boosted DNA is examined using a technique called gel
electrophoresis. Here, DNA fragments are separated based on their size and charge,
creating a DNA profile.
• DNA COMPARISON: The DNA profile from the sample is matched with profiles
from other individuals, like suspects or victims, to see if there's a match.

Figure: Methodology of DNA fingerprinting

Fluorescence in-situ hybridization (FISH)

• FISH is applied to provide specific localization of genes on chromosomes. Rapid

diagnosis of trisomies and microdeletions is acquired using specific probes.
• Usually, a denatured probe is added to a metaphase chromosome spread and incubated
overnight to allow sequence-specific hybridization.
• After washing off the unbound probe, the bound probe is visualized by its fluorescence
under UV light; thus, the site of the gene of interest is observed as in situ.
• This technique is used to check the cause of trisomies, microdeletion syndromes.

Figure: Methodology of Fluorescence in-situ hybridization

DNA Microarray
• DNA “chips” or microarrays have been used as possible testing for multiple mutations.
• In this technology, single DNA strands including sequences of different targets are
fixed to a solid support in an array format.
• On the other hand, the sample DNA or cDNA labeled with fluorescent dyes is
hybridized to the chip.
• Then using a laser system, the presence of fluorescence is checked; the sequences and
their quantities in the sample are determined.

Steps involved in DNA microarray

Sample collection:

▪ A sample can be any cell/tissue that we desire to conduct our study on.
▪ Generally, 2 types of samples are collected, i.e. healthy and infected cells, for
comparing and obtaining the results.

Isolation of mRNA:

▪ The extraction of RNA from a sample is performed by using a column or solvent like
phenol-chloroform.
▪ mRNA is isolated from the extracted RNA leaving behind rRNA and tRNA.
▪ As mRNA has a poly-A tail, column beads with poly-T tails are employed to bind
mRNA.
▪ Following the extraction, buffer is used to rinse the column inorder to isolate mRNA
from the beads.

Creation of labeled cDNA:

▪ Reverse transcription of mRNA yields cDNA.

▪ Both the samples are then integrated with different fluorescent dyes for the production
of fluorescent cDNA strands which allows to differentiate the sample category of the
cDNAs.

Hybridization:

▪ The labeled cDNAs from both the samples are placed on the DNA microarray which
permits the hybridization of each cDNA to its complementary strand.
▪ Then they are thoroughly washed to remove unpaired sequences.

Collection and analysis:

▪ Microarray scanner is used to collect the data.

▪ The scanner contains a laser, a computer and a camera. The laser is responsible for
exciting the fluorescence of the cDNA, generating signals.
▪ The camera records the images produced at the time laser scans the array.
▪ Then computer stores the data and yields results instantly. The data are now analyzed.
▪ The distinct intensity of the colors for each spot determines the character of the gene in
that particular spot.

Applications of DNA microarray technique:

▪ Drug discovery
▪ Study of functional genomics
▪ DNA sequencing
▪ Gene expression profiling
▪ Study of proteomics
▪ Diagnostics and genetic engineering
▪ Toxicological researches
▪ Pharmacogenomics and theranostics

Figure: Methodology of DNA microarray