Module-2 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.
CELLULOSE BASED WATER FILTERS

Cheap and simple decontamination of water could save millions of lives around the world as drinking
water contaminated with faecal bacteria causes life-threatening diseases such as cholera and other water-borne
diseases. It is estimated that 785 million people lack a basic drinking-water service (WHO, 2019).

Filtration is a common way to obtain pure drinking water by removing particles and microorganisms
based on size exclusion. The interest in the use of bio based filters for water purification has increased in recent
years; as such filters have the potential to be affordable, lightweight and biodegradable.

Wood-based cellulose pulp fibres are today used in disposable everyday filters, such as coffee filters and
air filters, and some of the first membranes for microfiltration were produced from cellulose nitrate and
cellophane in the late 1920s.

Cellulose-based water filters are filters made from cellulose, a carbohydrate polymer found in plant cell
walls. They are used to remove impurities and contaminants from water and are an alternative to traditional
synthetic polymer filters. Cellulose filters can effectively remove particles, pathogens, and other contaminants
from water, making it safer and more potable. Cellulose can be sourced from wood pulp, agricultural residues,
or other plant materials. The raw material is treated with chemicals (like alkalis) to remove lignin and
hemicellulose, leaving purified cellulose.

Properties of cellulose based water filter

 High Porosity: Cellulose-based water filters have a high porosity structure, which allows them to
efficiently remove impurities and contaminants from water.
 Biodegradability
 Cost-effective: affordable than traditional synthetic polymer filters
 Renewable resource: made from a renewable resource, cellulose
 Good mechanical strength: allowing them to maintain their structure and perform effectively over time.
 Chemical resistance: resistant to most chemicals, including acids and bases
 Large surface area: Cellulose-based water filters have a large surface area, which enhances their
filtration capabilities and reduces the frequency of filter replacement.

Additional features of cellulose that makes its suitable as water filters include:
 Cellulose being the most abundant biopolymer in nature has high biocompatibility, and contains several
functional groups exhibiting various advantages. Cellulose can be prepared into several derivatives
including nanomaterials such as
 Cellulose nanocrystals (CNCs),
 Cellulose Nano fibrils (CNFs) etc
 They can proceed into various forms such as membranes, sheets, papers, foams, aerogels, and filters.
  They can be applied for water remediation via removing water pollutants including Heavy metal ions,
namely uranyl, arsenic, copper, cadmium, chromium, lead and mercury owing to the presence of
different functional groups.
 Cationic cellulose or cellulose-based nanocomposites are widely used for the adsorption of anionic
pollutants e.g. nitrate and fluoride.
 Nano cellulose-based materials were reported for the adsorption of other pollutants such as
pharmaceuticals, cosmetic products, drugs, organic dyes and antibiotics
 Cellulose-based materials can be also used for removing microorganisms being active as membranes or
antibacterial agents, using antibacterial metal nanoparticles both silver nanoparticles (AgNPs) and
copper nanoparticles (CuNPs). Impregnating cellulose paper with AgNPs and CuNPs, have known to
inactivate bacteria during percolation through the filter.
 Another alternative method to physically remove bacteria from water, while keeping the filter pore size
larger than bacteria, is to use positively charged filters that adsorb negatively charged bacteria onto the
surfaces of the filters.

Construction of cellulose-based water filters

 Cellulose Material Selection: The type of cellulose material used in the water filter will depend on the
desired properties such as strength, porosity, and chemical resistance. Common cellulose materials
include paper, cotton, and wood fibers.
 Cellulose Preparation: The cellulose material is prepared by cutting it into small pieces, washing it to
remove impurities, and drying it for use.
 Cellulose Layer Formation: The cellulose material is formed into a layer by either stacking it or
compacting it using heat and pressure.
 Filter Medium Attachment: The cellulose layer is attached to a filter medium such as a mesh or a
support structure to provide stability and increase the filter surface area.
 Chemical Treatment: The cellulose layer may be chemically treated to modify its properties, such as
increasing its hydrophilicity or adding antimicrobial agents.
 Housing Assembly: The filter medium is assembled into a housing that provides a means to attach it to
a water source and to collect the filtered water.
 Filter Testing: The completed filter is tested to ensure that it meets the desired specifications, such as
filtration efficiency and flow rate.
Note: This is a general outline, the exact process may vary depending on the specific requirements of the water
filter and the type of cellulose material used.

Cellulose material used in house hold water filters

 The cellulose material commonly used in household water filters is cellulose acetate. Cellulose acetate is
a synthetic form of cellulose that has properties such as good chemical resistance, high porosity, and high flow
rate, making it suitable for use in household water filters. Additionally, cellulose acetate is also a low-cost
material, making it accessible for use in household applications.

BIOPLASTICS

Plastic is the third most commonly used petroleum derivative in the world; each year 200 million tons of
plastic are consumed on the planet. It comes from a non-renewable source (petroleum), it is contaminating and
non-biodegradable (it can take more than 1000 years to decompose).

Over the last several years, production of polymers from renewable resources has shown significant
growth. Some of the plastics produced from renewable resources such as vegetable oil, corn, and pea starch
have been synthesized by microbes and are known as bioplastics.

The prefix ‘bio’ in bioplastics can mean several things: the monomers were derived from renewable
resources (biomass) and then polymerized through chemical mechanisms; the polymer was extracted from
biomass; the polymer or the plastic is biodegradable or nonbiodegradable (note that the processing of a polymer
into its plastic product can affect the original biodegradability); the material is produced through biological
processes; or a combination of these. The use of ‘bioplastics’ for fossil-derived degradable plastics is
discouraged.

Its development is driven by current demands to replace fossil fuel–based polymers. Limitation in fossil
fuel resources, price volatility, impact on the environment, and waste disposal problems are some of the main
reasons for this shift toward bio-based plastics.

These are 100% degradable, equally resistant and versatile, already used in agriculture, textile industry,
medicine and, over all, in the container and packaging market.

POLYLACTIC ACID (PLA)

Polylactic Acid (PLA) is a biodegradable and bio-based plastic made from corn starch, sugarcane, or
other natural resources. PLA is an aliphatic (Aliphatic (or non-aromatic) polyesters are degradable because the
ester groups in their backbones can be easily cleaved by hydrolysis or enzymatic activity). It is a homopolymer
and is the most price-competitive synthetic bioplastic. PLA is typically made through the polycondensation of
lactic acid, which can be derived from the fermentation of sugars, or through the ring-opening polymerization of
lactide, the cyclic dimer of lactic acid. PLA can be optically clear and has found use as a replacement for
polyolefin films or polystyrene foams, including incorporation into single-use items and also in packaging,
disposable tableware, and 3D printing.

However, it's important to note that while PLA is biodegradable in industrial composting facilities, it
may not break down in the environment as quickly as advertised and may still have negative impacts on wildlife
and ecosystems if not properly disposed of.

Properties of PLA as bioplastic

 Biodegradable: PLA can be broken down by microorganisms in industrial composting facilities,

reducing waste in landfills.
 Renewable: PLA is derived from renewable resources such as corn starch or sugarcane, reducing
dependence on finite petroleum resources.
 Clear/Transparent: PLA has a clear and transparent appearance, making it suitable for packaging
applications.
 Heat-resistant: PLA has a relatively low melting temperature and is not recommended for high heat
applications, but it can maintain its shape and stability up to 60°C.
 Biocompatible: PLA is non-toxic and biocompatible, making it suitable for food packaging and medical
devices.
 Stiffness and Strength: PLA has good stiffness and strength, but not as strong as traditional petroleum-
based plastics.
 Printability: PLA is commonly used in 3D printing due to its good printability and ease of use.
Engineering applications of PLA bioplastic
 Automotive parts: PLA is used for the production of biodegradable automotive parts such as air ducts
and headlamp covers.
 Electronic Devices: PLA is used for the production of biodegradable components electronic devices
such as smartphones and laptops.
 Aerospace: PLA is used for the production of biodegradable parts in aerospace applications, such as
insulation and cable management.
 Sporting Goods: PLA is used for the production of biodegradable sporting goods such as golf tees and
fishing lures.
 Construction: PLA is used for the production of biodegradable insulation and soundproofing materials.
 Agricultural Equipment: PLA is used for the production of biodegradable parts in agricultural equipment
such as seed trays and greenhouse film.
 Medical Equipment: PLA is used for the production of biodegradable components in medical equipment
such as diagnostic equipment and hospital beds.

POLYHYDROXYALKANAOTAES (PHAs)
Polyhydroxyalkanoates (PHAs) are a class of biodegradable and biocompatible linear polyesters produced
intracellularly by organisms as carbon and energy reserves (bio-based) with similar physicochemical properties
to the conventional plastics. Instead of chemical synthesis, these polyesters are produced by fermenting raw
vegetable materials with a series of bacterial strains, Pseudomonas and Ralstonia strains, as well as algae.These
fermenting raw vegetable materials with a series of bacterial strains, Pseudomonas and Ralstonia strains, as
well as algae of their cell volume.They are also biodegradable and biocompatible, which makes them the next-
generation eco-friendly materials.

PHAs are

 Resistant to water and moisture and present a high range of in-use temperature and low oxygen and
water vapor permeability.
 The material properties of PHAs can be tuned by varying the repeat unit chain length, the side chain
functionalities and co-monomer composition, resulting in (co)polymers such as rigid and brittle poly (3-
hydroxybutyrate) (P3HB) or the softer and more flexible poly (3-hydroxybutyrate-co-3-
hydroxyhexanoate) (PHBH). Their desirable mechanical properties and good O2 and CO2 barrier
properties indicate that PHAs would be suitable replacements for bulk packaging materials such as
polyethylene and polypropylene.Most PHAs degrade faster than PLA, which makes them attractive for
applications in which biodegradation is desired.

Engineering applications of PHA bioplastic

 Packaging: used in various forms of packaging such as food containers, beverage cups, and clamshell
containers.
 Medical Devices: it is biocompatible and can be used in the manufacture of medical devices such as
sutures, implants, and drug delivery systems.
 Textiles: used in the production of biodegradable textiles, as well as for the production of biodegradable
composites for use in construction and furniture.
 Agricultural Mulch Films: PHA is used in the production of biodegradable mulch films for agriculture to
reduce soil erosion and conserve moisture.
 Consumer Goods: PHA is used in the production of various consumer goods, such as toys, phone cases,
and water bottles.
 Automotive Parts: PHA is used for the production of biodegradable automotive parts such as air ducts
and headlamp covers.

Advantages of bioplastics:

 They reduce carbon footprint

 They providing energy savings in production
 They do not involve the consumption of non-renewable raw materials
 Their production reduces non-biodegradable waste that contaminates the environment
 They do not contain additives that are harmful to health, such as phthalates or bisphenol A.
 They do not change the flavour or scent of the food contained

Nucleic acids in vaccines and diagnosis:

Nucleic acids, including DNA and RNA, have a wide range of applications in various fields such as medicine,
biotechnology, forensic science, agriculture, and environmental science.
 Gene Therapy: Introduction, alteration, or removal of genes within a patient's cells to treat disease. Used
to treat genetic disorders like cystic fibrosis, hemophilia, and certain cancers by correcting defective
genes or introducing new genes to fight diseases.
 Production of Vaccines: DNA Vaccines and RNA Vaccines.
 Recombinant DNA Technology: Genetic Engineering by Manipulates DNA to produce genetically
modified organisms (GMOs) for agriculture, medicine, and industry.
 Protein Production: Produces therapeutic proteins like insulin, growth hormones, and monoclonal
antibodies.
 Gene Synthesis: Creates synthetic genes for research, industrial biotechnology, and therapeutic
applications.
 DNA Profiling/ fingerprinting: Analyzes specific regions of DNA (short tandem repeats, STRs) to
identify individuals. Used in criminal investigations, paternity testing, and identification of human
remains.
 Bioremediation: Uses genetically engineered microorganisms to degrade environmental pollutants.
Cleans up oil spills, heavy metals, and other toxic substances in soil and water.
 Diagnostics: detecting pathogens through the identification of their genomic sequences
Nucleic acids in vaccines
DNA Vaccines
1. Plasmid DNA Encoding Antigen
o Contains the gene for the antigen of the pathogen.
2. Delivery into Host Cells
o Delivered using methods like electroporation or viral vectors.
3. Cellular Uptake
o Host cells take up the plasmid DNA.
4. Transcription
o DNA is transcribed into mRNA in the nucleus.
5. Translation
o mRNA is translated into the antigen protein in the cytoplasm.
6. Immune Response
o Antigen is presented on cell surface, triggering an immune response.
Eg: influenza virus, hepatitis B virus, human immunodeficiency virus, rabies virus, lymphocytic chorio-
meningitis virus, malarial parasites and mycoplasmas.
ADVANTAGES:
 Efficacy: DNA vaccines have been shown to be highly effective in preventing infections in both animal
and human trials.
 Long-lasting protection: DNA vaccines can stimulate a strong and long-lasting immune response, which
means that they can provide protection for extended periods of time.
 Ease of administration: DNA vaccines are easy to administer, as they can be given via injection or even
delivered orally, which can be particularly useful in areas where access to medical facilities is limited.
 Cost-effective: DNA vaccines are relatively inexpensive to produce compared to traditional vaccines,
which can make them more accessible in areas where resources are limited.
 Reduced risk of side effects: DNA vaccines do not contain live virus particles, which means that they
are generally safer and have a lower risk of side effects compared to traditional vaccines.

mRNA Vaccines
1. mRNA Encoding Antigen
o Contains the mRNA for the antigen of the pathogen.
2. Encapsulation in Lipid Nanoparticles (LNPs)
o Protects mRNA and facilitates delivery into cells.
3. Delivery into Host Cells
o LNPs deliver mRNA into the cytoplasm.
4. Translation
o mRNA is directly translated into the antigen protein in the cytoplasm.
5. Immune Response
o Antigen is presented on cell surface, triggering an immune response.
Eg: COVID19 vaccines, such as the Pfizer–BioNTech and Moderna vaccines
ADVANTAGES: mRNA vaccines, such as those developed for COVID-19 by Pfizer-BioNTech and Moderna,
offer several advantages over traditional vaccine platforms. Here are some key advantages:
 Rapid Production: mRNA vaccines can be developed and produced much faster than traditional
vaccines.
 Scalability: mRNA vaccines can be produced quickly and in large quantities
 No Risk of Infection: mRNA vaccines do not use live virus, so there is no risk of causing the disease in
the vaccinated individual.
 Non-integrating: mRNA does not integrate into the host genome
 Strong Immune Response: mRNA vaccines have been shown to elicit strong and effective immune
responses
 Customization: mRNA vaccines can be easily modified to address new variants of viruses

Nucleic acids in diagnosis:

Several infectious diseases caused by microorganisms, including protozoans, bacteria and viruses are
threats to public health. In order to control outbreaks, emergence, and reemergence of these infectious diseases,
diagnosis, correct identification, treatment, and notification of pathogenic agents are necessary.
Classical clinical microbiological diagnostics for protozoan, virus and bacteria rely on microscopic
examination with different staining methods, culture isolation etc. These standard diagnostic methods are very
useful. However, these diagnostic techniques are unsuitable for several microorganisms presenting fastidious
growth characteristics, low morphological and physiological specificity. Nucleic acid-based diagnostics are
pivotal in not only in identifying pathogens, but also genetic mutations, and other biomarkers associated with
diseases.
1. Polymerase Chain Reaction (PCR): PCR is a widely used molecular biology technique to amplify DNA
sequences, making it possible to detect and analyze small amounts of DNA. Steps in PCR
o Denaturation: DNA is heated to separate its strands.
o Annealing: Primers bind to the target DNA sequence.
o Extension: DNA polymerase extends the primers, creating a copy of the target sequence.
Applications:
 In Medical Diagnostics: to detect bacterial and viral pathogens (e.g., HIV, COVID-19).
 Genetic Disorders: Identifying mutations or genetic markers associated with diseases.
 Forensic Science: in DNA Profiling/ fingerprinting
 Research: Cloning and Sequencing: Amplifying DNA for further genetic analysis or cloning.
Advantages
  Sensitivity: Can detect very small amounts of DNA.
 Specificity: Targeted amplification of specific DNA regions.
 Speed: Results can be obtained in a few hours.

2. Reverse Transcription PCR (RT-PCR): RT-PCR is an essential tool in molecular biology used to amplify
RNA. It is a technique that combines reverse transcription of RNA into DNA with subsequent amplification
of DNA using PCR, i.e.,
Reverse Transcription: RNA is converted into complementary DNA (cDNA) using the enzyme reverse
transcriptase.
PCR Amplification: The cDNA is then amplified through standard PCR processes involving
denaturation, annealing, and extension.
Applications
 Infectious Disease Diagnostics: Detects RNA viruses such as SARS-CoV-2, HIV, and influenza.
 Gene Expression Analysis: Measures mRNA levels to study gene expression patterns.
 Cancer Research: Identifies specific RNA markers associated with cancer.
Advantages
 Sensitivity: Capable of detecting low levels of RNA.
 Specificity: Highly specific to target RNA sequences.
 Quantitative Analysis: When combined with quantitative PCR (qPCR), it can provide precise
quantification of RNA levels.

3. Next-generation sequencing (NGS)

Next-generation sequencing (NGS) is a type of DNA sequencing technology that uses parallel sequencing of
multiple small fragments of DNA to determine sequence.
 "High-throughput" technology has allowed a dramatic increase in the speed (and a decrease in the cost)
at which an individual's genome can be sequenced.
 Massive parallel sequencing
 Sequencing of entire genomes or specific regions of DNA and RNA.
Mechanism
1. Library Preparation: DNA/RNA is fragmented and adapters are added.
2. Sequencing: Fragments are sequenced simultaneously in a massively parallel manner.
3. Data Analysis: Sequenced data is assembled and analyzed to identify genetic variations.
Applications
1. Genomics: Whole-genome sequencing to study genetic variations and evolutionary biology.
2. Medical Diagnostics: Identifying mutations associated with diseases, including cancer.
3. Infectious Disease: Detecting and characterizing pathogens.
4. Personalized Medicine: Tailoring treatments based on individual genetic profiles.
Advantages
 High Throughput: Can sequence millions of fragments simultaneously.
 Speed: Rapid processing of large amounts of genetic data.
 Detail: Provides comprehensive genetic information, including rare variants

PROTEINS IN FOOD PRODUCTION

 Proteins are large biological molecules composed of one or more amino acid chains. These biopolymers
display high nutritional value, stabilization, elasticity, water binding ability, emulsification, gelation, and
foaming ability, as well as capacity to protect cells, tissues, and organisms.
Many plant and animal proteins could be used as a source of material for production and design of
protein-based structures. The use of animal proteins could bring various advantages such as low toxicity of the
degradation end products and high absorbability.
Sources of Proteins in Food Production
1. Animal Proteins:
o Meat: Beef, pork, poultry, and fish are primary sources of animal proteins used in various food
products.
o Dairy: Milk, cheese, yogurt, and whey are rich in proteins like casein and whey protein.
o Eggs: A versatile source of high-quality protein used in baking, sauces, and as an emulsifier.
2. Plant Proteins:
o Legumes: Beans, lentils, peas, and soybeans are significant sources of plant proteins.
o Cereals and Grains: Wheat, rice, oats, and quinoa contribute to protein content in foods.
o Nuts and Seeds: Almonds, chia seeds, and hemp seeds are high in protein and used in various food
products.
Functional Roles of Proteins in Food Production
1. Texturizing Agents:
o Gelation: Proteins like gelatin and casein can form gels, providing structure to products like
yogurt, cheese, and gelatin desserts.
o Emulsification: Proteins help stabilize emulsions, such as in mayonnaise, salad dressings, and
ice cream.
o Foaming: Egg whites and whey protein can create foams, important in products like meringues,
mousses, and whipped toppings.
2. Nutritional Enhancement:
o Fortification: Adding proteins to foods to enhance their nutritional profile, such as protein bars,
shakes, and fortified cereals.
o Dietary Supplements: Protein powders and shakes for athletes and individuals needing higher
protein intake.
o Nutritional value: Proteins are an important source of essential amino acids and provide
nutritional value in food systems. They contribute to the body's growth, repair, and
maintenance processes, making proteins a vital component of a balanced diet
3. Flavor and Color Development:
o Maillard Reaction: Proteins react with reducing sugars during cooking, leading to the
development of complex flavors and browning in products like bread, roasted meats, and coffee.
4. Binding Agents:
o Meat Processing: Proteins act as binders in processed meats like sausages, nuggets, and
meatballs, helping to hold ingredients together and improve texture.
o Water-binding capacity: Proteins have water-binding properties, which help improve the
moisture retention and juiciness of food products.
5. Texture modification: Proteins can contribute to the textural properties of food products by forming a
network that influences viscosity, elasticity, and mouthfeel. This property is particularly significant in
products like bakery goods, pasta, and meat analogs, where proteins play a crucial role in defining
the desired texture and bite
Some of the common applications of proteins in the food industry:
A. Dairy Industry: Cheese production, Yogurt and fermented dairy products, Milk and Whey processing,
Almond milk, soy milk, and pea protein milk serve as dairy substitutes.
B. Baking Industry
C. Juice Industry
D. Brewing Industry
E. Meat Processing
F. Meat Substitutes: Products like tofu, tempeh, and seitan are traditional plant-based proteins. Modern
alternatives include products made from pea protein, soy protein, and other plant sources to mimic the
texture and flavor of meat.

LIPIDS: BIODIESEL
Fuel demand and global warming have intensified research interest in the field of sustainable biofuels
energy to replace fossil fuels. Biodiesel has gained increasing attention due to the generation of alternate,
nontoxic, and renewable sources of liquid fuel as well as have advantages of high combustion efficiency, low
aromatic, low sulfur content, and highly biodegradable compared to other petrochemical fuels.
Food waste is recognized as an organic-rich resource consisting of protein (5%-10%), lipid (10%-40%),
and carbohydrate (20%-45%), and also it contain nutrients such as phosphate, and fatty acid so that it can be
used by means of valorization for conversion into biodiesel.
 Biodiesel is defined as long-chain fatty acid alkyl esters obtained from vegetable oils or animal fats by the
transesterification process.
 It is also known as fatty acid methyl ester or FAME.
 In the transesterification process, vegetable oils and fats (or triglycerides) from animals are converted into
esters, where alcohol reacts with fatty acids to form mono alkyl esters.


Pretreatment: Food waste contains minor quantities of lignin; therefore mild pretreatment is required.
Pretreatment is used to eliminate contamination and other particles or reduce the water content and acidity
value. It also enhances the transesterification process. The pretreatments are categorized as
 Physical: In physical pretreatment, extraction of biodiesel from the substrate can take place by applying
pressure or heat.
 Chemical pretreatment: is a process of extracting biodiesel from waste using chemicals (acid or base).
 Mechanical pretreatment is an effective method for releasing the organics from biomass through
mechanical means. Ultrasonic treatment is an efficient technique to improve the mass transfer rate
between immiscible liquid phases.
  Biological pretreatment is a process of treating a substance by biological means to achieve biodiesel,
through microalgae species like Chlorella vulgaris, Microcystis aeruginosa, Botryococcus braunii,
Dunaliella tertiolecta, and Chlorella prothothecoides or yeast Yarrowia lipolytica.
Transesterification is the process of converting mono-, di-, and triglycerides into FAME, known as biodiesel.
Transesterification indicates vegetable oil or fat reactions with an alcoholic group in the presence of a catalyst.
The catalyst can be a base, acid, or an enzyme. Methanol is the most widely used alcohol in the
transesterification process due to its low cost, and it has advantages such as producing clean fuel with a lower
molecular weight and viscosity.
There are two major conversion techniques involved in the transesterification reaction, namely
 Chemical (acidic and basic catalytic conversion) and
 Biological techniques [enzymatic (lipase) or bacterial or fungal conversion].
The transesterification process is carried out by two methods in biodiesel production:
(a) chemical-catalyzed method:
 Acid-catalyzed: The transesterification process carried out through acids is known as acid-catalyzed
transesterification. These types of acid-catalyzed transesterification are best suited for biodiesel
production from organic substrates. H2SO4 and H2O3S are preferable acids
 Base-catalyzed transesterification: A transesterification process that involves the application of alkali
or base as catalyst is known as alkaline- or base catalyzed transesterification. MgO, Al2O3, mixed
oxides of CaO, MgO, Li4SiO4, and Na2SiO3 are some of the alkaline-based catalysts used in the
transesterification process
(b) Enzyme-catalyzed method: Enzymatic transesterification has gained increasing attention because it
gives a high-quality product, and is easily separable, reusable, and has less of an impact on environment.
Lipase, an enzyme, is a significant biocatalyst which can be used in reactions such as hydrolysis,
acidolysis, alcoholysis, or transesterification and aminolysis.
The advantages of enzymatic catalyzed transesterification over chemical transesterification are mild
reactions, easy recovery, catalyst being recyclable, no inhibitory products, high product quality, no
saponification, a lower alcohol-to-oil ratio, and complete free fatty acids.

LIPIDS IN DETERGENTS PRODUCTION

Lipids have several properties that make them suitable as cleaning agents, including their ability to
emulsify and dissolve grease and oils. Additionally, lipids can form micelles, which are tiny spherical structures
that can surround and trap dirt particles, making it easier to remove them. The production of detergents from
lipids primarily involves converting natural fats and oils into soap and other surfactants.

Synthetic detergents often involve lipid-derived intermediates, such as fatty alcohols and fatty acids,
which are chemically modified to enhance their cleaning properties.

i. Hydrolysis of Triglycerides: Fats and oils (Triglycerides) are hydrolyzed to produce fatty acids and
glycerol.
Triglyceride + Water → Glycerol + Fatty Acids
ii. Fatty Alcohol Production: Fatty acids are hydrogenated to produce fatty alcohols.
Fatty Acid+H2→Fatty Alcohol
iii. Sulfonation and Sulfation: Fatty alcohols are sulfonated or sulfated to produce anionic surfactants.
Fatty Alcohol+SO3→Fatty Alcohol Sulfate

Types of Lipids Used in Detergents:

  Fatty Acids: These are often derived from natural fats and oils. Common sources include animal fats
and plant oils such as coconut oil, palm oil, and soybean oil.
 Triglycerides: These are esters derived from glycerol and three fatty acids. They can be found in
natural oils and fats.
 Phospholipids: Although less common in commercial detergents, they can be used in specialized
cleaning products for their superior emulsifying properties.

Functions of Lipids in Detergents:

 Surfactants: Lipids, especially fatty acids, are key components in the synthesis of surfactants.
Surfactants reduce the surface tension of water, allowing it to spread and wet surfaces more
effectively, enhancing the detergent’s ability to remove dirt and grease.
 Emulsifiers: Lipids help in the formation of emulsions, stabilizing mixtures of oil and water. This is
particularly important in removing oily stains from fabrics and surfaces.
 Foaming Agents: Many lipid-derived surfactants contribute to foam production, which is important for
consumer perception of cleaning effectiveness, even though foam itself does not necessarily enhance
cleaning performance.

Advantages of Lipid-Based Detergents:

1. Biodegradability: Fatty acid-based soaps and detergents are generally more biodegradable than
petroleum-based products.
2. Skin-Friendly: Natural lipid-derived detergents are less likely to cause skin irritation.
3. Sustainable Sources: Utilizing renewable resources such as vegetable oils and animal fats supports
sustainability.
ENZYMES IN FOOD PROCESSING
Enzymes are proteins which catalyze chemical reactions selectively as part of essential life processes such
as digestion, respiration and metabolism in organisms. For centuries, human beings have exploited the catalytic
efficiency of these ubiquitous biomolecules for food processing, especially in the preparation of beer, wine,
cheese and bread. Enzymes are generally extracted from edible plants and the tissues of animals. Certain
enzymes also are produced by microorganisms like bacteria, yeasts, and fungi. Enzymes play a critical role in
food processing by catalyzing biochemical reactions that improve the quality, texture, taste, and shelf-life of
food products. Some key applications of enzymes in various sectors of the food industry include:
1. Baking Industry
 Amylases: Break down starches into sugars, providing fermentable sugars for yeast, improving
dough handling, and enhancing bread volume and texture.
 Proteases: Improve dough extensibility and reduce mixing times, which is crucial for producing
high-quality bread and baked goods.
 Lipases: Help in dough conditioning and improve the crumb structure and shelf-life of baked
products.
2. Dairy Industry
 Rennet (Chymosin): Used in cheese-making to coagulate milk, separating it into curds and whey.
 Lactase: Breaks down lactose into glucose and galactose, aiding in the production of lactose-free
dairy products.
 Lipases: Enhance the flavor of cheeses by breaking down milk fats.
2. Brewing Industry
  Amylases: Convert starches in grains into fermentable sugars, essential for beer production.
 Proteases: Improve the stability and clarity of beer by breaking down proteins that can cause
haze.
 Glucanases: Break down glucans to improve the filtration process and prevent issues with
viscosity.
3. Fruit and Vegetable Processing
 Pectinases: Used to break down pectin, which helps in the clarification of fruit juices and wine,
and in increasing juice yield.
 Cellulases and Hemicellulases: Aid in the breakdown of cell walls, improving the extraction of
juices and purees.
 Amylases: Enhance the sweetness and texture of fruit products by breaking down starches.
4. Meat Processing
 Proteases (Papain, Bromelain, Ficin): Tenderize meat by breaking down muscle proteins, making
it more palatable and easier to chew.
5. Oil Extraction
 Lipases: Improve oil yield and quality by breaking down triglycerides into free fatty acids and
glycerol.
 Cellulases and Hemicellulases: Aid in breaking down the cell walls of oilseeds, enhancing oil
extraction efficiency.
6. Confectionery
 Invertase: Converts sucrose into glucose and fructose, preventing crystallization in candies and
improving texture.
 Amylases: Used in the production of high-fructose corn syrup (HFCS) by breaking down starch
into glucose.
Advantages of Using Enzymes in Food Processing
 Specificity: Enzymes act on specific substrates, leading to desired reactions without unwanted
by-products.
 Mild Conditions: They operate under mild conditions of temperature and pH, preserving the
nutritional and sensory qualities of food.
 Efficiency: Enzymes speed up reactions, making processes faster and more efficient.
 Sustainability: Enzymes can reduce the need for chemical additives and harsh processing
methods, making food processing more environmentally friendly.

ENZYMES IN DETERGENT FORMULATION

Enzymes are used in biological detergents for cleaning laundry and crockery, due to their ability to
break down various stains and soils effectively, improving cleaning performance and efficiency. Other than
efficient cleaning, using enzymes in detergents are sustainable and also save energy as they function in low
temperatures
Most of these detergents are created with a mix of 2 or more enzymes and can go up to 8 different
enzymes in a single formulation. These blends make the enzymes more tolerant of the other ingredients in the
detergent.
Enzyme detergent formulation usually comprises enzymes, surfactants, and builders.
 Enzymes: Alkaline proteases, amylases, and lipases help in cleaning by breaking down the dirt through
hydrolysis. Cellulases work on fabrics by hydrolysis of glycosidic bonds in fabrics that remove pilling,
 fuzz or lint attached to cotton microfibers. They also improve the softness and brightness of worn and
jaded clothes.
 Surfactants act by lowering the surface tension in liquid solution
 Builders act by providing alkalinity to the solution that prevents the dirt from reattaching to the fabric.
The most popular class of enzymes used in detergents is hydrolases, which remove stains from fat,
protein, and polysaccharide dirt. Some examples of stains include food, grass, blood, animal and vegetable fat,
and dirt. The major types of hydrolases used in detergents include proteases, amylases, lipases, and mannanases.
Other classes of enzymes like cellulases and pectinases also have specific applications in detergent
formulations.
 Proteases: Break down protein-based stains such as blood, egg, grass, and human sweat. These enzymes
hydrolyze peptide bonds in proteins, converting them into smaller peptides and amino acids, which can
be easily removed during washing.
 Amylases: Degrade starch-based stains like those from pasta, potatoes, and baby food. They hydrolyze
starch molecules into soluble sugars, facilitating their removal from fabrics.
 Lipases: Target fat and oil stains, including grease, butter, and cosmetic oils. Lipases break down
triglycerides into fatty acids and glycerol, making it easier to wash away these stains.
 Cellulases: Remove cellulose-based stains and rejuvenate cotton fabrics. These enzymes degrade
cellulose micro fibrils on the fabric surface, reducing fuzz and pills, thereby improving fabric
smoothness and appearance.
 Mannanases: Effective against food-based stains containing mannans, such as sauces, ice cream, and
processed foods. Mannanases break down mannans into simpler sugars, aiding in their removal from
textiles.
 Pectinases: Break down pectin, a component of fruit stains and certain plant-based stains. They
hydrolyze the pectin in stains, helping to release these substances from the fabric.

Advantages of Using Enzymes in Detergents

 Enhanced Cleaning Performance: Enzymes target specific types of stains, breaking them down into
smaller, more soluble molecules that are easier to wash away.
 Lower Washing Temperatures: Enzymes are effective at lower temperatures, allowing for energy
savings and better fabric care.
 Reduced Environmental Impact: Enzymes are biodegradable and reduce the need for harsh chemicals,
making detergents more environmentally friendly.
 Fabric Care: Enzymes help maintain fabric integrity by preventing the build-up of fuzz and pills,
extending the life of garments.
 Efficiency: Enzymes work synergistically with other detergent components, enhancing the overall
cleaning power without increasing the amount of detergent needed.
 Cost-effective

ENZYMES IN TEXTILE PROCESSING

Enzymes play a vital role in the textile processing industry by providing environmentally friendly alternatives to
traditional chemical processes. Their use improves efficiency, reduces water and energy consumption, and
enhances the quality of the final products.
Enzymes are safely used in several textile processes like
 Desizing: Removal of size (starch) from the warp yarns after weaving.
 Enzymes Used: Amylases for efficient removal of size without damaging the fabric, reduced water
consumption, and lower energy requirements.
 Scouring: Removal of natural impurities such as pectin, waxes, and oils from cotton.
Enzymes Used: Pectinases, cellulases for enhanced fabric absorbency, better dyeing properties, and
reduced use of harsh chemicals.
 Biopolishing: Improving fabric surface by removing micro fibrils and fuzz.
Enzymes Used: Cellulases for smoother fabric surface, reduced pilling, and enhanced fabric brightness
and softness.
 Bleaching: Whitening of fabric by removing natural color.
Enzymes Used: Laccases, catalases for reduced chemical use, milder processing conditions, and lower
environmental impact.
 Denim Finishing: Creating a worn and faded look in denim fabrics.
Enzymes Used: Cellulases, laccases for reduced use of pumice stones and chemicals, lower water
consumption, and softer denim.
 Degumming: Removal of sericin from silk.
Enzymes Used: Proteases for gentle removal of sericin, preserving silk luster and strength.

Types of Enzymes in Textile Processing

1. Amylases: Remove starch-based sizes from fabrics.
2. Cellulases: Improve the texture and appearance of cellulosic fabrics.
3. Pectinases: Break down pectin substances in cotton.
4. Proteases: Modify protein-based fibers and remove protein stains.
5. Laccases: Oxidize phenolic compounds.
6. Catalases: Decompose hydrogen peroxide.

Advantages of Using Enzymes in Textile Processing

 Environmentally Friendly: Enzymes are biodegradable and reduce the need for harsh chemicals, leading
to less environmental pollution.
 Energy Efficient: Enzymatic processes typically occur at lower temperatures and neutral pH, reducing
energy consumption.
 Improved Fabric Quality: Enzymes work specifically on target substrates, preserving the integrity of
fibers and enhancing the final product's quality.
 Water Savings: Enzyme-based processes often require less water, contributing to water conservation
efforts in the textile industry.
 Selective Action: Enzymes are highly specific, allowing for precise and controlled processing.

ENZYMES IN BIOSENSORS FABRICATION

A biosensor is a device that uses biological molecules to detect and measure the presence of various
substances. It typically consists of three main components:
 Bioreceptor: A biological material (like enzymes, antibodies, or nucleic acids) that specifically interacts
with the substance being detected.
 Transducer: Converts the interaction between the Bioreceptor and the substance into a measurable
signal.
 Electronic System: Processes and displays the signal in a user-friendly format.

Biosensors can be classified based on the type of Bioreceptor like enzymes, antibodies, or nucleic acids
used. Enzymes are extensively utilized in the fabrication of biosensors due to their high specificity, catalytic
activity, and ability to operate under mild conditions. These properties make them ideal for detecting
various biological and chemical substances.

Types of Enzyme-Based Biosensors

 Electrochemical Biosensors: Measure changes in electrical properties (current, voltage, impedance)

due to the enzymatic reaction.
Eg: Glucose oxidase, lactate oxidase, urease.
Used in Blood glucose monitoring, lactate sensing in athletes, detection of urea in clinical
diagnostics.
 Optical Biosensors: Detect changes in optical properties (absorbance, fluorescence, luminescence)
resulting from the enzymatic reaction.
Eg: Horseradish peroxidase, luciferase.
Used in Environmental monitoring, food qualities control, clinical diagnostics.
 Thermal Biosensors: Measure the heat change associated with the enzymatic reaction.
Eg: Various enzymes depending on the analyte.
Used in Metabolic studies, monitoring of fermentation processes.

Applications of Enzyme-Based Biosensors

 Medical Diagnostics:
o Glucose Sensors: For monitoring blood glucose levels in diabetic patients using glucose
oxidase.
o Lactate Sensors: For monitoring lactate levels in athletes and critically ill patients using
lactate oxidase.
o Cholesterol Sensors: For measuring cholesterol levels using cholesterol oxidase.
 Environmental Monitoring:
o Detection of Pollutants: Enzymes like urease and acetylcholinesterase are used to detect
pollutants such as heavy metals and pesticides.
o Monitoring of Water Quality: Enzymes can detect contaminants like nitrates and phosphates.
 Food Quality Control:
o Detection of Pathogens: Enzymes like horseradish peroxidase are used in biosensors to detect
foodborne pathogens.
o Monitoring of Freshness: Enzyme-based sensors can detect spoilage markers in food
products.
 Industrial Applications:
o Fermentation Monitoring: Enzymatic sensors monitor the levels of key metabolites in
fermentation processes.
o Bioprocess Control: Real-time monitoring of substrates and products in bioprocesses.

Advantages of Enzyme-Based Biosensors

1. High Specificity: Enzymes provide high specificity for their substrates, ensuring accurate detection.
2. Sensitivity: Enzymatic reactions can amplify the signal, providing high sensitivity.
3. Mild Conditions: Enzymes operate under mild conditions, which is beneficial for the stability of the
sensor and the analyte.
4. Rapid Response: Provides quick results compared to traditional laboratory methods.
5. Portability: Many biosensors are compact and can be used on-site or in the field.