MODULE 1

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 Module - 1
BIOMOLECULES AND THEIR APPLICATIONS (QUALITATIVE)
Carbohydrates (cellulose-based water filters, PHA and PLA as bioplastics), Nucleic acids (DNA
Vaccine for Rabies and RNA vaccines for Covid19, Forensics – DNA fingerprinting), Proteins
(Proteins as food – whey protein and meat analogs, Plant based proteins), lipids (biodiesel,
cleaning agents/detergents), Enzymes (glucose-oxidase in biosensors, lignolytic enzyme in bio-
bleaching).

BIOMOLECULES AND THEIR APPLICATIONS:

What is a biomolecule?
Biomolecule, also called biological molecule, any of numerous substances that are produced by
cells and living organisms.

Biomolecules have a wide range of sizes and structures and perform a vast array of functions. The
four major types of biomolecules are carbohydrates, lipids, nucleic acids, and proteins. Among
biomolecules, nucleic acids, namely DNA and RNA, have the unique function of storing an
organism’s genetic code—the sequence of nucleotides that determines the amino acid sequence of
proteins, which are of critical importance to life on Earth.

There are 20 different amino acids that can occur within a protein; the order in which they occur
plays a fundamental role in determining protein structure and function. Proteins themselves are
major structural elements of cells.

They also serve as transporters, moving nutrients and other molecules in and out of cells, and as
enzymes and catalysts for the vast majority of chemical reactions that take place in living
organisms. Proteins also form antibodies and hormones, and they influence gene activity.

Carbohydrates, which are made up primarily of molecules containing atoms of carbon, hydrogen,
and oxygen, are essential energy sources and structural components of all life, and they are among
the most abundant biomolecules on Earth. They are built from four types of sugar units—
monosaccharides, disaccharides, oligosaccharides, and polysaccharides.

Lipids, another key biomolecule of living organisms, fulfill a variety of roles, including serving
as a source of stored energy and acting as chemical messengers. They also form membranes, which
separate cells from their environments and compartmentalize the cell interior, giving rise to
organelles, such as the nucleus and the mitochondrion, in higher (more complex) organisms.
Examples include cytidine, uridine, adenosine, guanosine, and thymidine. Nucleosides that are
phosphorylated become nucleotides. Apart from serving as a structural unit of nucleic acids,
nucleotides may also serve as sources of chemical energy (e.g. adenosine triphosphate or ATP).

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The main applications of biomolecules are:
The biomolecules may involve in several processes such as energy storage (carbohydrates),
catalyzing the biochemical reactions (hormones), storing/transmitting the genetic codes
(RNA/DNA), or altering biological and neurological activities (neurotransmitter/hormones).

COURTESY: GEEKSFORGEEKS

CARBOHYDRATES:

What are carbohydrates?

Carbohydrates, or carbs, are sugar molecules. Along with proteins and fats, carbohydrates are
one of three main nutrients found in foods and drinks. Body breaks down carbohydrates into
glucose. Glucose, or blood sugar, is the main source of energy for your body's cells, tissues, and
organs. Glucose can be used immediately or stored in the liver and muscles for later use.

There are three main types of carbohydrates:

1) Sugars: They are also called simple carbohydrates because they are in the most basic form.
They can be added to foods, such as the sugar in candy, desserts, processed foods, and regular
soda. They also include the kinds of sugar that are found naturally in fruits, vegetables, and milk.
Starches. They are complex carbohydrates, which are made of lots of simple sugars strung
together. Your body needs to break starches down into sugars to use them for energy. Starches
include bread, cereal, and pasta. They also include certain vegetables, like potatoes, peas, and corn.
Fiber. It is also a complex carbohydrate. Your body cannot break down most fibers, so eating
foods with fiber can help you feel full and make you less likely to overeat. Diets high in fiber have
other health benefits. They may help prevent stomach or intestinal problems, such as constipation.
They may also help lower cholesterol and blood sugar. Fiber is found in many foods that come
from plants, including fruits, vegetables, nuts, seeds, beans, and whole grains.
Just like starches, cellulose is another best example for carbohydrates.

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2) Cellulose: a complex carbohydrate, or polysaccharide, consisting of 3,000 or more glucose
units.
It is extremely abundant, easily renewable, and biodegradable. Due to inter- and intramolecular
hydrogen bonding between the hydroxyl groups of the neighboring cellulose chains, cellulose is
insoluble in water, despite being hydrophilic, and is difficult to dissolve with common organic
solvents.

Taking benefit of these advantages of cellulose, we have a best application of cellulose, that is,
Cellulose-based water filters.

The interest in the use of biobased filters for water purification has increased in recent years, as
such filters have the potential to be affordable, lightweight and biodegradable. Research has been
focused on creating biobased membranes for micro- and ultrafiltration from cellulose nanofibrils
(CNFs).
Filters based on cellulose pulp fibers do usually have large pores that facilitate water percolation
but they do not sufficiently remove bacteria through size exclusion; other techniques are therefore
needed to achieve a bacteria-reducing effect. Several groups have addressed this issue by
incorporating antibacterial metal nanoparticles into cellulose-based water filters; both silver
nanoparticles (AgNPs) and copper nanoparticles (CuNPs) are known to have good antibacterial
effects.
An alternative method 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.

This allows negatively charged particles much smaller than the filter pore size to be efficiently
removed from water and this is an interesting approach for removing bacteria from water without
adding any toxic chemicals or reducing the flow by reducing the pore size. Both Gram-positive
and Gram-negative bacteria have a negative net surface charge on the cell envelope, due to
peptidoglycans, liposaccharides and proteins in the cell wall, and this makes their removal non-
selective and efficient for most types of bacteria.
Methods used for the same are:

 LBL [Layer By Layer] MODIFICATION

 NITROGEN ANALYSIS
 SEM [ Scanning Electron Microscope]
 FLOW RATE FOR FREE FLOW FILTRATION
 BACTERIAL REMOVAL EFFICIENCY OF FILTRATION
 FILTRATION OF NATURAL WATER SAMPLES
 FLOURESCENCE MICROSCOPY

Cellulose filter papers are versatile and diverse tools for microfiltration, that work by trapping
particulates within a random matrix of cellulose fibers. Cellulose filter papers can be categorized
as quantitative or qualitative, depending on their application.

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 Cellulose based nanomaterials for water treatment

POLY LACTIC ACID [PLA] AND POLYHYDROXYALKANOATES [PHA]:

Just like, how we introduced biodegradable water treatment plant from cellulose, we also have a
replacement for toxic, non-biodegradable plastics which we are using in our daily life. That is,
Bioplastics.
Bioplastics are one type of plastic which can be generated from natural resources such as starches
and vegetable oils. 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 by where it ends up at the end of its cycle life. Bio plastics are said to be
biodegradable if they are broken down with the effect of the right environmental conditions and
microbes which in turn use them as a food source. The bioplastics are considered compostable if
within 180 days, a complete microbial assimilation of the fragmented food source takes place in a
compost environment.
Based upon this, we have PHA and PLA.

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 Production of biopolymers

PLA is both: biobased and biodegradable under industrial composting conditions (at a high
temperature, around 58 °C). Because of its good mechanical properties, processability,
renewability, and non-toxicity, PLA is considered today as one of the most commercially
promising bioplastics. When compared with most other biodegradable polymers, PLA has better
durability, transparency, and mechanical strength.

PHAs are a significant polymer family that are 100% bio-based and bio-degradable. PHAs are
microbiologically produced polyesters that have tunable physical and mechanical properties. This
is accompanied by low environmental impact due to their biodegradability and non-toxicity nature.
Therefore, they are promising candidates for a sustainable future manufacturing. Ranging from
brittle thermoplastics to gummy elastomers, PHAs' properties can be altered by the selection of
bacteria, fermentation conditions, and substrate. Due to their flexible properties, PHAs can
eventually substitute PP, polyethylene (PE), and polystyrene (PS), which are the main polymers
of today's global polymer market.

PLA PHA

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 STEPS IN PRODUCING BIOPLASTIC FROM PHA AND PLA

NUCLEIC ACID:

Nucleic acids are biopolymers, macromolecules, essential to all known forms of life.They are
composed of nucleotides, which are the monomers made of three components: a 5-carbon sugar,
a phosphate group and a nitrogenous base. The two main classes of nucleic acids are
deoxyribonucleic acid (DNA) and ribonucleic acid (RNA). If the sugar is ribose, the polymer is
RNA; if the sugar is the ribose derivative deoxyribose, the polymer is DNA.

Nucleic acids are naturally occurring chemical compounds that serve as the primary information-
carrying molecules in cells and make up the genetic material. Nucleic acids are found in abundance
in all living things, where they create, encode, and then store information of every living cell of
every life-form on Earth. In turn, they function to transmit and express that information inside and
outside the cell nucleus to the interior operations of the cell and ultimately to the next generation
of each living organism. The encoded information is contained and conveyed via the nucleic acid
sequence, which provides the 'ladder-step' ordering of nucleotides within the molecules of RNA
and DNA. They play an especially important role in directing protein synthesis.
Strings of nucleotides are bonded to form helical backbones—typically, one for RNA, two for
DNA—and assembled into chains of base-pairs selected from the five primary, or canonical,
nucleobases, which are: adenine, cytosine, guanine, thymine, and uracil. Thymine occurs only in
DNA and uracil only in RNA.
The two main nucleic acids are DNA and RNA,which is the fundamental unit of any living
organisms. Based on these factors, there are many applications for the same, some of which are
explained below:

1) DNA VACCINE FOR RABIES:

Rabies is a preventable viral disease most often transmitted through the bite of a rabid animal. The
rabies virus infects the central nervous system of mammals, ultimately causing disease in the brain
and death. Most rabies cases reported to the Centers for Disease Control and Prevention (CDC)
each year occur in wild animals like bats, raccoons, skunks, and foxes, although any mammal can
get rabies.

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A DNA vaccine, using a pCl-neo plasmid encoding the glycoprotein gene of a Mexican isolate of
rabies virus, was developed to induce long-lasting protective immunity against rabies virus in dogs.
The worldwide incidence of rabies and high rates of therapy failure, despite availability of effective
vaccines indicate the need for timely and improved prophylactic approaches. DNA vaccination
based on optimized formulation of lysosome-targeted glycoprotein of the rabies virus provides
potential platform for preventing and controlling rabies. A range of parameters including physical,
physiological, clinical, immunological, hematological along with histopathology profiles of target
organs was monitored to assess the impact of vaccination.

There were no observational adverse effects despite high dose administration of the DNA vaccine
formulation. Thus, this study indicates the safety of next generation of vaccines as well as
highlights their potential application.

DNA Vaccine production

COURTESY: BABIUK

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2) RNA VACCINES FOR COVID-19:
Coronavirus disease (COVID-19) is an infectious disease caused by the SARS-CoV-2 virus.
Messenger RNA, or mRNA technology, instructs cells to make a protein that generates an immune
response in the body, thus producing the antibodies that provide protection against a disease.
It is the basis for the Pfizer/BioNTech and Moderna COVID-19 vaccines being used by
governments worldwide, and in the UN-supported COVAX global vaccine solidarity initiative.
Messenger ribonucleic acid (mRNA) is a molecule that provides cells with instructions for making
proteins. mRNA vaccines contain the instructions for making the SARS-CoV-2 spike protein. This
protein is found on the surface of the virus that causes COVID-19.

The mRNA molecule is essentially a recipe, telling the cells of the body how to make the spike
protein. COVID-19 mRNA vaccines are given by injection, usually into the muscle of the upper
arm. After the protein piece is made, the cell breaks down the instructions and gets rid of them.
The mRNA never enters the central part (nucleus) of the cell, which is where our DNA (genetic
material) is found. Your DNA can't be altered by mRNA vaccines.

The cell then displays the protein piece on its surface. Our immune system recognizes that the
protein doesn't belong there and begins building an immune response and making antibodies.

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3) FORENSIC-DNA FINGERPRINTING:

DNA fingerprinting, also called DNA typing, DNA profiling, genetic fingerprinting, genotyping,
or identity testing, in genetics, method of isolating and identifying variable elements within the
base-pair sequence of DNA (deoxyribonucleic acid).

The procedure for creating a DNA fingerprint consists of first obtaining a sample of cells, such as
skin, hair, or blood cells, which contain DNA. The DNA is extracted from the cells and purified,
the DNA was then cut at specific points along the strand with proteins known as restriction
enzymes. The enzymes produced fragments of varying lengths that were sorted by placing them
on a gel and then subjecting the gel to an electric current (electrophoresis): the shorter the fragment,
the more quickly it moved toward the positive pole (anode).

The sorted double-stranded DNA fragments were then subjected to a blotting technique in which
they were split into single strands and transferred to a nylon sheet. The fragments underwent
autoradiography in which they were exposed to DNA probes—pieces of synthetic DNA that were
made radioactive and that bound to the minisatellites. A piece of X-ray film was then exposed to
the fragments, and a dark mark was produced at any point where a radioactive probe had become
attached. The resultant pattern of marks could then be analyzed.
The DNA testing process is comprised of four main steps, including extraction, quantitation,
amplification, and capillary electrophoresis.

DNA fingerprinting is a laboratory technique used to determine the probable identity of a person
based on the nucleotide sequences of certain regions of human DNA that are unique to individuals.
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.

CUORTESY: LISA H CHADWICK

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 COURTESY: FORENSIC SCIENCE

DNA FINGERPRINTING

So, this is all about nucleic acids, considering the two main nucleic acids, that is, DNA and RNA,
discussing about some of the most important applications of these nucleic acids.

PROTEINS:

Protein is found throughout the body—in muscle, bone, skin, hair, and virtually every other body
part or tissue. It makes up the enzymes that power many chemical reactions and the hemoglobin
that carries oxygen in your blood.
Talking more about proteins based on biomolecule concept, Proteins are large biomolecules and
macromolecules that comprise one or more long chains of amino acid residues. Proteins perform
a vast array of functions within organisms, including catalysing metabolic reactions, DNA
replication, responding to stimuli, providing structure to cells and organisms, and transporting
molecules from one location to another. Proteins differ from one another primarily in their
sequence of amino acids, which is dictated by the nucleotide sequence of their genes, and which
usually results in protein folding into a specific 3D structure that determines its activity.
Proteins are assembled from amino acids using information encoded in genes. Each protein has its
own unique amino acid sequence that is specified by the nucleotide sequence of the gene encoding
this protein. The genetic code is a set of three-nucleotide sets called codons and each three-
nucleotide combination designates an amino acid.

The process of synthesizing a protein from an mRNA template is known as translation. The mRNA
is loaded onto the ribosome and is read three nucleotides at a time by matching each codon to its
base pairing anticodon located on a transfer RNA molecule, which carries the amino acid

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corresponding to the codon it recognizes. The enzyme aminoacyl tRNA synthetase "charges" the
tRNA molecules with the correct amino acids. The growing polypeptide is often termed the nascent
chain. Proteins are always biosynthesized from N-terminus to C-terminus.
Proteins are the chief actors within the cell, said to be carrying out the duties specified by the
information encoded in genes. With the exception of certain types of RNA, most other biological
molecules are relatively inert elements upon which proteins act. Proteins make up half the dry
weight of an Escherichia coli cell, whereas other macromolecules such as DNA and RNA make
up only 3% and 20%, respectively. The set of proteins expressed in a particular cell or cell type is
known as its proteome.

PROTEIN AS FOOD:

Protein is a key part of any diet. The average person needs about 7 grams of protein every day for
every 20 pounds of body weight. Because protein is found in an abundance of foods, many people
can easily meet this goal. However, not all protein “packages” are created equal. Because foods
contain a lot more than protein, it’s important to pay attention to what else is coming with it.
Animal-based foods (meat, poultry, fish, eggs, and dairy foods) tend to be good sources of
complete protein, while plant-based foods (fruits, vegetables, grains, nuts, and seeds) often lack
one or more essential amino acid.

Whey protein is a mixture of proteins isolated from whey, the liquid material created as a by-
product 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. Whey protein is commonly marketed as a protein supplement, and various health
claims have been attributed to it.

Whey is left over when milk is coagulated during the process of cheese production, and contains
everything that is soluble from milk after the pH is dropped to 4.6 during the coagulation process.
It is a 5% solution of lactose in water and contains the water soluble proteins of milk as well as
some lipid content. Processing can be done by simple drying, or the relative protein content can
be increased by removing the lactose, lipids and other non-protein materials.
The primary usage of whey protein supplements is for muscle growth and development. Eating
whey protein supplements before exercise will not assist athletic performance, but it 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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Meat analogues find raising interest of many consumers who are looking for indulgent, healthy,
low environmental impact, ethical, cost-effective, and/or new food products. High moisture
extrusion cooking enables the production of fresh, premium meat analogues that are texturally like
muscle meat from plant or animal proteins. The appearance and eating sensation are similar to
cooked meat while high protein content offers a similar nutritional value. This article focuses on
plant-based meat analogues and covers process and product-related aspects including ingredients
and structure formation, flavor, taste and nutritional value, postextrusion processing, packaging
and shelf life, consumer benefits, and product-related environmental impacts.
Meat analogues, can be defined as products that mimic meat in its functionality, bearing similar
appearance, texture, and sensory attributes to 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.

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The factors that lead to this shift is due to low fat and calorie foods intake, flexitarians, animal
disease, natural resources depletion, and to reduce greenhouse gas emission. Currently, available
marketed meat analog products are plant-based meat in which the quality (i.e., texture and taste)
are similar to the conventional meat. The ingredients used are mainly soy proteins with novel
ingredients added, such as mycoprotein and soy leghemoglobin.

COURTESY: MEENAKSHI SINGH

PLANT BASED PROTEINS:

Plant protein is simply a meaningful food source of protein which is from plants. This group can
include pulses, tofu, soya, tempeh, seitan, nuts, seeds, certain grains and even peas. Pulses are a
large group of plants, which include chickpeas, lentils, beans (such as black, kidney and adzuki
beans) and split peas.

Plant proteins are highly nutritious – not only as good sources of protein, but also because they
provide other nutrients such as fibre, vitamins and minerals. Our intake of fibre tends to be too
low, however by incorporating certain plant proteins into your diet, such as pulses, peas and nuts,
you can easily boost your fibre intake.
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: (1)potential
health benefits associated with increased intake of plant-based diets; (2) consumer concerns
regarding adverse health effects of consuming diets high in animal protein (e.g., increased
saturated fat); (3) increased consumer recognition of the need to improve the environmental
sustainability of food production; (4) ethical issues regarding the treatment of animals; and (5)
general consumer view of protein as a “positive” nutrient (more is better). While there are health

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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.

LIPIDS:

Discussing about another important biomolecule, lipids, are a broad group of naturally occurring
molecules which includes fats, waxes, sterols, fat-soluble vitamins (such as vitamins A, D, E and
K), monoglycerides, diglycerides, phospholipids, and others. The functions of lipids include
storing energy, signaling, and acting as structural components of cell membranes. Lipids have
applications in the cosmetic and food industries, and in nanotechnology.

Lipids may be broadly defined as hydrophobic or amphiphilic small molecules; the amphiphilic
nature of some lipids allows them to form structures such as vesicles, multilamellar/unilamellar
liposomes, or membranes in an aqueous environment. Biological lipids originate entirely or in part
from two distinct types of biochemical subunits or "building-blocks": ketoacyl and isoprene
groups.Using this approach, lipids may be divided into eight categories: fatty acyls, glycerolipids,
glycerophospholipids, sphingolipids, saccharolipids, and polyketides (derived from condensation
of ketoacyl subunits); and sterol lipids and prenol lipids (derived from condensation of isoprene
subunits).

A biological membrane is a form of lamellar phase lipid bilayer. The formation of lipid bilayers is
an energetically preferred process when the glycerophospholipids described above are in an
aqueous environment.This is known as the hydrophobic effect. In an aqueous system, the polar
heads of lipids align towards the polar, aqueous environment, while the hydrophobic tails minimize
their contact with water and tend to cluster together, forming a vesicle; depending on the
concentration of the lipid, this biophysical interaction may result in the formation of micelles,
liposomes, or lipid bilayers. Other aggregations are also observed and form part of the
polymorphism of amphiphile (lipid) behavior.
Some of the applications are: Within the body, lipids function as an energy reserve, regulate
hormones, transmit nerve impulses, cushion vital organs, and transport fat-soluble nutrients. Fat
in food serves as an energy source with high caloric density, adds texture and taste, and contributes
to satiety.

Lipid obtained from food waste was used as a potential feedstock for biodiesel production using
both a chemical catalyst and a biocatalyst. Base (KOH) catalyzed transesterification of the lipid
allowed a 100% conversion of biodiesel in 2 h; whereas, Novozyme-435 yielded 90% biodiesel in
24 h. So lipids are having a main application in bidiesel production.
Biodiesel demonstrates an animal fat-based or vegetable oil diesel fuel, including long-chain alkyl
(methyl, ethyl, or propyl) esters. Biodiesel is generally made by esterifying lipids (e.g., soybean
oil, vegetable oil, and animal fat (tallow)) with an alcohol generating fatty acid esters. Biodiesel is
suggested to be utilized in standard diesel engines and is thus well defined from the vegetable and
waste oils used to fuel converted diesel engines. Biodiesel can be used singly or blended with
gasoline in any proportions. Biodiesel blends can also be utilized as heating oil.

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 Biodiesel production
COURTESY: WEI JIN

Lipids also has an interesting application in cleaning agents. Considering the case of

DETERGENTS:

The hydrophobic end of the phospholipid bilayer stays away from the water. This avoids the
dissolution of cell membrane in water. But the detergent can bind to the hydrophobic end of the
cell membrane and form a solution with water, thus breaking the cell membrane barrier.
Detergent monomers solubilize membrane proteins by partitioning into the membrane bilayer.
With increasing amounts of detergents, membranes undergo various stages of solubilization. The
initial stage is lysis or rupture of the membrane.
While lipids also have the same general structure as detergents—a polar hydrophilic head group
and a nonpolar hydrophobic tail— lipids differ from detergents in the shape of the monomers, in
the type of aggregates formed in solution, and in the concentration range required for aggregation.

COURTESY: FAZAL SHAIK

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ENZYMES:

Are another important biomolecule, which are proteins that help speed up metabolism, or the
chemical reactions in our bodies. They build some substances and break others down. All living
things have enzymes. Our bodies naturally produce enzymes.
The six kinds of enzymes are hydrolases, oxidoreductases, lyases, transferases, ligases and
isomerases.

Enzymes perform the critical task of lowering a reaction's activation energy—that is, the amount
of energy that must be put in for the reaction to begin. Enzymes work by binding to reactant
molecules and holding them in such a way that the chemical bond-breaking and bond-forming
processes take place more readily.
Due to their high specificity, simplicity, and scalability, enzyme-based biosensors represent a fast,
precise, and continuous monitoring of analytes. Additionally, the high specificity of enzymes
enhances the ability to detect lower analyte concentration limits. So enzymes are used in
biosensors.

Biosensors are employed in applications such as disease monitoring, drug discovery, and detection
of pollutants, disease-causing micro-organisms and markers that are indicators of a disease in
bodily fluids (blood, urine, saliva, sweat).
Various types of biosensors being used are enzyme-based, tissue-based, immunosensors, DNA
biosensors, and thermal and piezoelectric biosensors. There are wide variety of enzymes used in
biosensors. One such enzyme is glucose oxidase, mainly in amperometric glucose biosensor.

GLUCOSE OXIDASE IN BIOSENSORS:

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

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 COURTESY: RESEARCH GATE

LIGNOLYTIC ENZYME IN BIO BLEACHING:

Ligninolytic enzymes play a key role in degradation and detoxification of lignocellulosic waste
in environment. The major ligninolytic enzymes are laccase, lignin peroxidase, manganese
peroxidase, and versatile peroxidase.
Ligninolytic fungi and enzymes (i.e., laccase, manganese peroxidase, and lignin peroxidase) have
been applied recently in the production of second-generation biofuels.

White-rot fungi are the main producers of lignin-oxidizing enzymes. These fungi secrete a number
of oxidative enzymes and some hitherto unknown substances (mediators) into their environment
together effecting a slow but continuous degradation. The most important lignin-oxidizing
enzymes are lignin peroxidases, manganese peroxidases and laccases. Lignin peroxidase and
manganese peroxidase appear to constitute a major component of the ligninolytic system.

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COURTESY:SHARDESH KUMAR CHAURASIA

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