Biology for Engineers Module-II

Module-2
APPLICATION OF BIOMOLECULES
Syllabus:
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

Introduction

Biomolecules are organic molecules present in the body and they support many
biological/chemical processes of living beings. They are being recognized to perform numerous
functions, and hence, a lot of research is being dedicated to explore their benefits to maximum
extent. Biomolecules can be tailored to make them suitable for the proposed applications and can
be synthesized and produced in large scales using advanced techniques. Biomolecules find their
applications in cosmetics, therapeutic, pharmacy, textile, biofuels, pulp and paper industries, waste
water treatment and various others. Some of them are discussed here

Water filters

Cellulose
Cellulose is a complex carbohydrate consisting of 3,000 or more glucose units. In nature, cellulose
is found in many plants’ cell walls. In fact, cellulose is the most abundant organic polymer on
Earth - a long chain polymer of glucose. It is the core “ingredient” in both cotton and wood.
Cellulose is non digestible by humans but it is the main food for herbivorous animals (like cow).
Cellulose and its derivatives are used to make paperboard, paper, adhesives, moisture proof
coatings etc. They are also used in water filters

Carbohydrates in cellulose-based water filters production

Water pollutants such as arsenic, chromium, lead, mercury, fluoride etc cause widespread illness
and reduces the quality of life. Hence water purification is highly required.
There are several types in water purification namely sedimentation, filtration, coagulation,
precipitation, adsorption etc and nano materials are used in these methods. Among several nano

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materials, cellulose is promising. Cellulose based materials are used to purify water through
different processes. Here we discuss two of them

Coagulation: When Chemicals like cellulose nano crystals (CNC), ferric sulphate, ferric chloride
(called coagulants) are added to water, they bind with the pollutants due to opposite charges present
on them. As a result a large particle is formed. This is known as coagulation. Coagulant is added
and circulated continuously throughout the water and finally large particles are removed by means
of filtration. Natural organic matter, dissolved organic carbon, inorganic pesticides, suspended
solids etc can be removed from this method. Saw dust based CNC is suitable to remove Ni2+ and
Cd2+ ions

Adsorption: It is a physical process in which dissolved pollutants in water (called adsorbate) are
attracted and attached to the surface of something larger called adsorbent.(it is similar to a magnet
on a refrigerator). Adsorption occurs due to electrostatic force, Van der Waals force, hydrogen
bond formation and ion exchange. Adsorption can remove many kinds of pesticides, chlorinated
hydrocarbons, gasoline, industrial solvents and disinfection by-products and heavy metals ions

When polluted water is passed through paper filter made up of cellulose fibers, Ag+, Cu2+, Pb2+,
Hg2+ As3+ ions are attracted to negatively charged groups present in cellulose materials and thereby
they can be removed. However these filters do not remove bacteria because of large pores size in
the cellulose fibers. If anti-bacterial nano particles like silver or copper nano particles are
incorporated in these pores then bacteria can also be removed from the water

Cellulose based materials such as CNC, CNF (cellulose nano fibrils) are used to purify water by
means of adsorption method.

Coagulation

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Adsorption

Bioplastics

Conventional plastic is produced from petroleum or natural gas. It is the third most commonly
used petroleum derivative in the world. Plastic does not biodegrade. It can take up to 1,000 years
to break down. When it is discarded, it builds up in the environment until it reaches a crisis point.
This pollution affects marine life, damages soil and poisons groundwater and cause serious health
impacts. In this regard bioplastics are very important

Bioplastics are plastics materials produced from renewable biomass sources, such as vegetable
fats and oils, corn starch, straw, woodchips, recycled food waste, sugar derivatives like starch,
lactic acid, cellulose etc.

PHA and PLA in bio-plastics production

There are two main forms of bioplastics on the market today: polylactic acids (PLA) and
polyhydroxyalkanoates (PHA).

PLA (polylactic acid)

PLA is typically made from the sugars in corn starch, cassava (genasu in kannada) or sugarcane.
It is biodegradable, carbon-neutral and edible.
First of all, corn kernels (seeds) are soaked in sulfur dioxide and hot water. This process breaks
the corn down into starch, protein, and fiber. The kernels are then ground up (mash) and the corn
oil is separated from the starch. Starch is made up of long chains of carbon molecules, similar to
the carbon chains in plastic from fossil fuels. It is mixed with citric acid in order to form a long-
chain polymer. This long chain is the building block for all plastics. This is a brief procedure to
produce PLA plastic

PLA can look and behave like polyethylene (used in plastic films, packing, and bottles),
polystyrene (Styrofoam and plastic cutlery) or polypropylene (packaging, auto parts, textiles).
PLA plastics are most commonly used for food packaging, cutleries and bottles, 3-D printing,
carpets, clothing, medical implants and devices etc

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Advantages of PLA
 PLA products are renewable and comparable replacement.
 PLA products come from bio-based materials like sugar starches
 PLA plastics are biodegradable
 Unlike other plastics, they do not emit toxic fumes when burnt.
 PLA is thermoplastic, so it is moldable and malleable when heated to its melting
temperature
 PLA products are freezer safe. Cups made up of PLA can withstand up to 2 000 F

PHA (polyhydroxyalkanoate)
PHA is produced by fermenting natural raw material such as sugar or lipids. First, bacteria are
fermented to grow a biopolymer within their casing. Once the desired yield is reached, the bacteria
are harvested from the fermentation broth (soup) to increase the concentration. The biopolymer is
then extracted from the bacteria cell, which can be done physically or chemically depending on
each production specifications. Finally, the lysate is extracted to form the biopolymer. PHA is
often more adaptable, biodegradable and less elastic than other plastics.

PHA is used for medical applications such as sutures, slings, bone plates, and skin substitutes,
single-use food packaging, straws etc

Advantages of PHA
 PHAs are fully biodegradable both in soil as well as in water
 PHAs are produced from renewable resources and hence are more sustainable option
compared to petroleum-based plastics
 The production of PHAs typically results in lower greenhouse gas emissions
 PHAs can be tailored to possess a wide range of properties, from being rigid and tough
to flexible and elastic.
 PHAs exhibit good thermal stability and hence used for higher temperature
applications
 Degradation products of PHAs are non-toxic

Vaccine

Edward Jenner created the world’s first successful vaccine in 1796 (for small pox). Since then
many vaccines have been designed for various infectious diseases. Vaccines have effectively
decreased the morbidity (illness) and mortality (death) across the large human population. They
have eradicated some diseases and restricted some diseases. However there are some challenges
and limitations for the vaccines. Scientists are trying to develop biomolecule based vaccines such
as nucleic acid vaccines. Nucleic acid-based vaccines are also known as naked DNA vaccines and
normally nucleic acid vaccines are injected into the muscle.

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Nucleic acid
Nucleic acid is a macromolecule that carries the genetic information of a cell and carries
instructions for the functioning of the cell. The two main types of nucleic acids
are deoxyribonucleic acid (DNA) and ribonucleic acid (RNA).

Nucleic acids in vaccines

A brief explanation of designing and working of Nucleic acid vaccines is as follows

In Nucleic acid vaccines, a genetic materials like DNA or RNA from disease causing virus or
bacteria (called pathogen) are used to stimulate an immune response against it. These materials
provides instructions for making the specific protein from the pathogen. Thus formed proteins are
called antigens (a foreign body). Once the DNA or RNA is inside the cell and it starts producing
antigens, these are then displayed on its surface, where they can be detected by the immune system,
triggering a response. This response includes production of killer T cells and B cells. T cells
destroy the infected cells and B cells produce antibodies which attack on invading bacteria. This
is how the nucleic acid vaccine works.
[Since many aspects of the immune response generated by DNA vaccines are not clearly
understood they are not used on humans so far. The field of DNA vaccination is developing
rapidly. Efforts are going on across the globe to produce DNA vaccine for cancer, TB, HIV and
many other diseases.]

Nucleic acids in diagnosis

Timely and accurate diagnosis of infectious diseases caused by bacteria viruses and many others
microorganisms (pathogens) helps in combating these pathogens and cure the disease. Many kinds
of diagnostics have been developed but they have certain limitations. Nucleic acid-based
diagnostics overcome the limitations of traditional diagnostic methods and become more widely
adopted in clinical tests. This technique is based on the amplification of genetic materials such as
DNA and RNA and hence detect the pathogens

Nucleic acid amplification techniques (NAAT) is a process of detecting pathogens through the
identification of their genomic sequences. There are two steps: sample pre-treatment and genetic
analysis. In sample pre-treatment step, pathogen nucleic acid is extracted and purified from a
biological sample (blood, urine, saliva, etc.). In the genetic analysis, a nucleic acid amplification
is done. Polymerase chain reaction (PCR) is a commonly used amplification method. PCR is a in-
vitro enzymatic reaction (in-vitro means tests done in labs) that allows the production of large
quantities of a specific fragment of DNA by means of DNA polymerase. When DNA (or RNA)
are replicated many times it is possible to detect minute traces of an organism present in the
specimen.

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NOTE: Nucleic acid sequence based amplification (NASBA), loop-mediated isothermal

amplification (LAMP) and displacement amplification (DA) are other different amplification
methods

Advantages:
 Nucleic acid–based methods are generally specific and highly sensitive
 It is useful for organisms that are difficult to culture or identify using other methods
 It is effective even if microorganisms are present in low numbers.
 It is very fast so that the patient can be placed on specific treatment immediately
 Diseases like viral fever, hepatitis, HIV, various types of cancer, Chromosome
abnormalities, Coagulation Factors, Cystic Fibrosis etc

DNA fingerprinting is another example for Nucleic acid diagnostics. It 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.

Food production

Proteins
Proteins are large and complex biomolecules that play many critical roles in the body. Enzymes,
hormones, blood protein, hemoglobin, kerratin etc are examples for proteins. In addition to having
many vital functions within the body, proteins perform different roles in our foods by adding
certain functional qualities to them

Proteins in food production

Protein is found both in animal-based foods (meat, poultry, fish, eggs, and dairy foods) and plant-
based foods (fruits, vegetables, grains, nuts, and seeds). Proteins and isolates proteins (protein
powders) are used in food industries for their functional properties like fat binding capacity,
emulsification (mixing of two different phases. ex. Oil and water), enhancement of flavor, foaming
ability and stability, adhesion, etc. They improve functional and nutritional properties of products.

Some examples are

 Enzymes are used in processing industries like starch, bakery, juice, beverages and dairy
where they act as catalysts and convert raw materials into improved food products
 Protein powders derived from whey, soy, or pea are added to sports drinks, nutritional
shakes and meal replacements to provide essential amino acids
 Protein powders are widely consumed as nutritional supplements for athletes,
bodybuilders and individuals

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 Ovalbumin, a major protein found in egg white acts as emulsifier and foaming agent. It is
used in food cakes. It gives foam when agitated and air bubbles trapped in the foam gives
the airy texture to the food cake
 Casein coagulate is a milk protein. It increases the thickness of yogurt when added
 Most cake recipes use eggs because the egg proteins help bind all the other ingredients
together into a uniform cake batter.
 Enzymes like lactase can be used to convert lactose into glucose or lactose-free milk
products.
 Enzymes like amylases contribute to the extension of shelf life of bakery products (like
bread and cake) by maintaining freshness, moisture content and softness.
 Lipolytic enzymes can be used to reduce the fat content in meat products.

Biodiesel

In recent years, biodiesel has been promoted as a renewable and environmentally preferable fuel
option to petroleum diesel. Biodiesel can be used in diesel engines alone or blended with actual
diesel

Lipids
Lipids are fatty or waxy compounds in the body and they are hydrophobic or insoluble in water,
Fats and oils, waxes, steroids etc. are examples for lipids. Cholesterol is also one example for the
lipid.

Lipids in biodiesel

Biodiesel is a liquid biofuel produced by reacting lipids with an alcohol. This also produces free
glycerol as impurity. Since glycerol forms harmful deposits in engines and damage emission
systems, it shall be limited to lowest possible value.

Raw materials for biodiesel

The raw materials for biodiesel production are vegetable oils, animal fats and short chain alcohols.
Vegetable oils include sunflower oil, soya bean oil, cotton seed oil, castor oil, ground nut oil, palm
oil etc. Methanol and ethanol are commonly used alcohols.

Production process
Biodiesel is produced through a trans-esterification reaction. This reaction converts an ester into a
mixture of esters of the fatty acids that makes up the oil (or fat). Biodiesel is obtained from the
purification of the mixture of fatty acid methyl esters (FAME). A catalyst is used to accelerate the
reaction. According to the catalyst used, trans-esterification can be basic, acidic or enzymatic. The
whole mechanism is represented as follows

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Here RCOOR/ is lipid (oil or fat), R//OH is alcohol and RCOOR//is biodiesel.

When methanol is used, the product of the reaction is a mixture of methyl esters and if ethanol is
used the product would be a mixture of ethyl esters. In both cases, glycerin will be the co-product
of the reaction. The mixture of esters and glycerin are separated. The ester mixtures are then
purified in order to remove contaminants such as water, excess alcohol and remnant catalysts

Advantages of biodiesel
 Renewable fuel, obtained from vegetable oils or animal fats.
 Low toxicity in comparison with diesel fuel.
 Degrades more rapidly than diesel fuel, minimizing the environmental consequences of
biofuel spills.
 Lower emissions of carbon monoxide, particulate matter, polycyclic aromatic
hydrocarbons, aldehydes.
 Lower health risk due to reduced emissions of carcinogenic substances.

Disadvantages of biodiesel
 Slightly higher fuel consumption due to the lower calorific value of biodiesel.
 Slightly higher nitrous oxide emissions than diesel fuel.
 Higher freezing point than diesel fuel. This may be inconvenient in cold climates.
 It is less stable than diesel fuel, and therefore long-term storage is not recommended.
 May degrade plastic and natural rubber gaskets and hoses when used in pure form

Lipids in detergents production

Soaps and detergents are the substances which possess the ability to remove dirt from surfaces
such as the human skin, textiles, and other solids when dissolved in water.

As we all know, majority of the substances dissolve in water and they are called hydrophilic (water
loving). But there are some substances which do not dissolve and they are called hydrophobic
(water-fear). For ex, oil, grease, dirt, soil, stains of color etc. do not dissolve in water. In order to

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remove them we use detergents. Detergents contain amphipathic lipid molecules that contain both
hydrophilic and hydrophobic groups. They have a hydrophilic group or polar group as head and a
long hydrophobic carbon chain or non-polar group as tail. When added to water, the polar group
forms hydrogen bonds with water molecules, while the hydrocarbon chains aggregate (gather
together) due to hydrophobic interactions. These properties allow detergents to be soluble in water.
In aqueous solutions, they form organized spherical structures called micelles (as shown Figure),
each of which contain several detergent molecules. Because of their amphipathic nature, detergents
are able to solubilize hydrophobic compounds such as oil, dirt, soil etc in water. They are then
removed by means of running water. For more effective cleaning, enzymes and surfactants are also
used. Lysolipids and fatty acids are two common lipids used in detergents

Applications of Enzymes

Enzymes
Enzymes are special kind of biomolecules which act as catalyst in various chemical reactions
taking place in our body. For ex, food digestion, liver function, muscle building, breathing etc
need the assistance of enzymes. They are used in many applications. Some of them are discussed
in brief here

Biosensors
A biosensor is a device used to measure biological or chemical reactions by generating signals
proportional to the concentration of an analyte in the reaction and hence detect disease-causing
micro-organisms and indicators of a disease in bodily fluids (blood, urine, saliva, sweat). A
biosensor consists of
 Analyte: A substance of interest that needs detection. Ex: glucose is an ‘analyte’ in a biosensor
designed to detect glucose.
 Bioreceptor: A molecule that specifically recognizes the analyte is known as a bioreceptor. Ex:
Enzymes
 Transducer: The transducer is an element that converts one form of energy into another. In a
biosensor the role of the transducer is to convert the bio-recognition event into a measurable signal
(signalization). Most transducers produce either optical or electrical signals

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 Electronics: It consists of complex electronic circuits where the signals are amplified and
converted into readable form
 Display: It is either computer display or LCD where output signal is displayed in numeric, graphic,
table or as an image depending on the requirements of the end user.

Enzyme biosensors

An enzyme biosensor is an analytical device that combines an enzyme with a transducer to produce
a signal proportional to target analyte concentration. The reactions catalyzed by the enzymes
produce the signal. Signals may be produced due to change in proton concentration (H+), release
or uptake of gases (like NH3, CO2, O2 etc), light emission, absorption or reflectance, heat emission
etc. The transducer converts this signal into a measurable signals (electrical, optical or thermal).
This signal can be further amplified, processed, or stored for later analysis.

Uses:
 Enzymatic biosensors are used to measure/detect glucose, lactate, alcohol, pH variation,
 Enzyme biosensors are used to detect traces of poisonous and harmful residues of
pesticides present in food grains, fruits and vegetables
 They are also used to detect ammonia and methane
 They may also be used to detect any pathogenic organisms present in meat, poultry, eggs,
and fish.
 Enzymatic sensors are used for scientific research and for process control in the food and
chemical industry

Advantages
 More specific
 Ease in miniaturization
 Robust
 Faster response
 Capable of operating with small sample volumes

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Lipids in Food processing

Microorganisms like bacteria, yeasts, and fungi have been used in food fermentation since ancient
times. Microbial enzymes produced from bacteria, yeasts, and fungi play a major role in food
industries because they are more stable than plant and animal enzymes. Many of these enzymes
find numerous applications in food sectors. The food processing mechanism by means of enzymes
involves one or many of the following reactions

 Oxidation-reduction in which oxygen and hydrogen are gained or lost

 Transfer of functional groups such as an amino group, acetyl group, or phosphate group
 Hydrolysis (addition of water)
 Removal of groups of atoms without hydrolysis
 Rearrangement of atoms within a molecule
 Joining of two molecules

Some of the enzymes generally employed in food industry are listed below:
Microbial enzyme Application
α-Amylase Bakery products, Bread quality improvement
Beer production
Glucoamylase
High glucose and high fructose syrups
Meat tenderization
Protease
Coagulation of milk
Cheese flavor development
Lipase
Cheddar cheese production
Animal feed
Cellulase
Clarification of fruit juice
Removal of bitter taste in fruit juice
Debittering enzymes
Wine aroma enhancement

Enzymes in detergent formulation

Enzymes are used as functional ingredients in detergents to clean laundry and dishes in an efficient
and energy saving manner. Proteases, lipases, amylases, cellulases are commonly used enzymes
in detergents. Many complex and stubborn stains such as chocolate, ice cream, baby food, desserts,
dressings, sauces, blood, grass, egg, animal & vegetable fat can be easily cleaned. The detergent
mechanism based on enzymes is as follows.

A detergent basically consists of surfactants, builders and enzymes. Soil and stains are removed
by mechanical action assisted them. Surfactants reduces the surface tension of water. Builders
increases the cleaning action by maintaining pH and softening hard water. Enzymes hydrolyze and

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solubilize substrate soils attached to fabrics or hard surfaces (e.g., dishes). The dirt is then removed
by water jet.

Advantages
 They impart greater softness and improve color brightness of worn cotton surfaces.
 They offer effective and effortless washing (time saving)
 In dish washer most of the energy is spent to heat the water. But enzyme based detergents
available today work at low temperatures (cold detergents) and hence lot of energy is
saved
 They are environmental friendly. They have replaced phosphate based detergents which
were causing harm to the environment
 Larger lumps of burnt-on and caked-on soils in vessels and frying pans can be easily
removed by these detergents
 Enzyme based detergents are compact

Enzymes in Textile processing

Enzymes are used increasingly in the textile processing industry. The specific textile enzymes used
are amylases, catalase, cellulases and laccase for removing the starch, degrading excess hydrogen
peroxide, textile bleaching, lignin-degrading, denim finishing (for jeans, shirts and jackets), bio-
polishing and decolorization

Advantages
 Enzymes can be used at room temperature and milder
conditions
 Quicken the process by accelerating the rate of reaction
 Help reduce wastage of water, energy, and time.
 The resultant finished products are of higher quality.
 They are non-toxic, biodegradable, non-polluting and safe
to use
 They give great finish to rayon, linen, cotton knits and
woven fabrics

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