11

Unit 11 Applications of Enzymes

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UNIT

APPLICATIONS OF
ENZYMES
Structure
11.1 Introduction 11.3 Summary
Objectives 11.4 Terminal Questions
11.2 Applications of Enzymes 11.5 Answers
Enzymes in Detergent Industry
Enzymes in Starch Industry 11.6 Suggested Readings
Enzymes in Baking Industry
Enzymes in Fruit juice, Brewing and
Wine Industry
Enzymes in Food Industry
Enzymes in Other Industries
Enzymes in Medicine/ Diagnostic
Enzymes
Enzyme Electrodes

11.1 INTRODUCTION
Enzymes have remarkable catalytic properties. You are now familiar with
several aspects of enzymes as illustrated in several previous units of this
course. As you know, enzymes speed up the rate of reactions, production
processes could be performed at a fraction of time at much lower temperature
and pressure. These advantages along with several others such as specific
catalytic activity, undesirable side reactions and their operability under mild
conditions make the enzymes highly desirable in comparison to other
chemical catalysts. Moreover, the usage of enzymes is not new. Role of
several enzymes especially in starch, cheese making, brewing and textile
industries has been carried out centuries before their biochemical aspect was
understood. With the advent of genetic engineering, there has been a
revolution in the field of enzyme technology. The enzymes present in number
of bacteria and fungi were found for many years by man but it was only in late
1970s isolation of enzymes was carried out and their commercial aspects
began to be explored. The separation, isolation and purification of enzymes
along with their immobilization have opened opportunities and challenges to
scientists in the field of enzyme technology as well as application of enzymes.
Now-a-days a wide range of enzymes are available on a much bigger scale
and a much lesser cost. As the whole area of enzymes is expanding, the
industrial and commercial application of enzyme is also increasing. The
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estimated market of enzymes in 2018 was approximately 4500 million dollars.
The present unit will give you an overview on several applications of enzyme in
different processes and industries.

Objectives
Enzymes are now finding wide spread applications. After studying this unit,
you will be able to:

 describe the usage of enzymes in detergent industry;

 illustrate the role of enzymes in food industry such as starch;
 state the role of enzymes in baking industry;
 explain the commercial viability of enzymes in wine and brewing
industries;

 determine the role of diagnostic enzymes; and

 enumerate several other applications of enzymes and their potential
commercial uses.

11.2 APPLICATIONS OF ENZYMES

As you know applications of enzymes are of immense value. An estimation of
world enzyme market in 2018 put the figure roughly in approximately 4500
million dollars. The effective catalytic properties of enzymes have already
promoted their introduction in several industrial products and processes.
Genetic manipulation, site directed mutagenesis, protein engineering, cloning
and genetic recombination along with several other techniques in
biotechnology have proved extremely useful to develop new version of
enzymes with better stability and catalytic efficiency. Enzymes are now tailor
made to suit the needs of medical and technical industry with vast range of
applications. Several enzymes are extensively used in textile, pulp, detergent,
beer, bakery and starch industries. Enzymes also have tremendous
therapeutic and diagnostic value. Now-a-days large scale production of
enzymes is also needed for research purposes.

Microbial sources are the ideal choice for protein isolation and purification.
Microorganisms are preferred over animal or plant as sources for enzyme
production. They can be easily grown in large numbers in bioreactors at
nominal cost using simple nutrients. They can also withstand extreme pH,
temperature or dissolved oxygen like conditions. Extracellular enzymes are a
better choice than the intracellular enzymes because of their ease of isolation
and purification. Do you know why? The isolation and purification of
extracellular enzymes is much simpler and cheaper. Microbial strains that
produce certain enzymes are the ideal choice or target for commercial viability.
The microorganisms can be easily screened and selected for producing the
desired enzyme. The procedure is economical too. Furthermore, it would be
advantageous if the organism does not produce any toxic compounds. Higher
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and better yield of enzyme without any production of unwanted toxic molecule
make the particular strain of microorganisms a better choice than other
sources of enzyme. Isolation and purification of a thermostable enzyme is
more convenient than other thermosensitive enzymes.

Though several enzymes are used in the industry using whole microbial
organism as a source of enzymes yet, the efficiency and efficacy of industrial
production processes can be enhanced using isolated enzymes. The useful
enzymes are separated, isolated and purified to increase the catalytic activity
and better yield of the product. The inherent disadvantages of these processes
also involve cost in separation and isolation as well as purification processes
of enzyme or inactivation of the enzyme. The thermophilic bacteria and fungi
are the ideal sources of enzymes for industrial or commercial use.

Other sources of enzymes are plant and animal tissues. Plants are excellent
sources of enzymes such as papain. Animal tissues contain several enzymes
such as lipases, esterases and proteases. However, there are several
limitations in isolating enzymes from plant and animal sources. Non-microbial
sources of enzymes, plant and animal tissues have damaging materials such
as phenolic compounds and endogenous enzyme inhibitors and proteases.
The quantities of enzyme in plant and animal tissues are limited and highly
varied in their distribution. With the advent of plant and animal tissue culture,
nowadays the possibility of producing large quantities of enzymes is being
explored. The constraining factor is their high cost. Several proteins such as
vaccines are being produced in large numbers using tissue culture techniques.
The methods available for isolation of proteins or enzymes are generally the
same. You will come across these techniques for isolation and purification of
biomolecules in several other courses of biochemistry. Presently 80 % of the
commercial enzymes are from microbial sources. The following table (Table
11.1) shows the sources of enzymes along with their relative contribution in the
commercial enzyme market.

Table 11.1: Source of enzymes and their contribution in commercial

enzyme market

Sources of enzyme Contribution (in %)

Fungi 60

Bacteria 24

Yeast 04

Streptomyces 02

Higher Animals 06

Higher Plants 04

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SAQ 1
Answer the following

a. Why microbes are considered as better sources of enzymes?

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b. Name two techniques used to develop new version of enzymes with better
stability and catalytic efficiency.

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c. Which source of enzyme will be better- extracellular enzyme or

intracellular enzyme? Why?

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11.2.1 Enzymes in Detergent Industry

Detergent industry is one of the largest single market for enzymes. Alkaline
proteases, amylases and lipases from microbial sources are being used
because of their stability at high temperature, alkaline pH and reduction of
agitation period. The dirt on clothes includes many compounds including
proteins, lipids and starch. Blood stains as well as stains contaminated with
milk or oils can be effectively and efficiently removed from the clothes by
detergent enzymes. These enzymes are proving to be cost effective and also
lead to less energy consumption. Enzymes in granulated forms as well as
liquid preparations are used in very small amounts in the detergent industry.
Some of the home detergents include amylase as well as protease enzyme as
these can be further used for laundering lot of dirty clothes and fabrics.
Industrial laundering is large and requires high level of cleanliness with minimal
usage of water. Newer versions of detergents include alkaline and
thermostable enzymes as fungal cellulase which generally removes small
fibers thereby restoring tensile strength of the fabric.

11.2.2 Enzymes in Starch Industry

Starch industry developed rapidly after 1990s. Starch containing crops such as
164 maize, potato and wheat are an important constituent of food consumed by the
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world’s population. Starch containing parts of the plant needs to be processed
to harvest the starch. Starch is used in different food products as glucose,
maltose, high fructose corn syrups and starch hydrolysates. Plant products
such as maize, potato and wheat are well known sources of starch for
commercial purposes. Several enzymes involved in processing of starch are
endoamylases, exoamylases, debranching enzymes and transferases. A
pictorial representation of the enzymes involved in starch processing and its
several products is shown in Fig. 11.1.

Starch

-Amylase

Dextrin Cyclodextrin

Fungal -Amylase

Amyloglucosidase Pullulanase
Pullulanase -amylase

Maltose Glucose High Syrup Maltose high

maltotriose fructose syrup

Glucose
Isomerase Crystallization

Fructose Syrup Crystalline sugar

Fig.11.1: Enzymes of starch industry

In the unit of carbohydrates, you might have come across starch, a polymer of
glucose units which are linked via -glycosidic linkage involving C1 and C4
hydroxyl groups. Generally, starch is present in the form of granules. These
granules differ widely in size and physical characteristics. Endoamylases
cleave -(1 4) glycosidic bonds present in amylase or amylopectin chain in
the starch. They are found in bacteria and archaea. Similarly, exoamylases
cleave exclusively -(14) and -(16) glycosidic bonds. They act on
external glucose residues of amylase or amylopectin to produce glucose.
Starch converting enzymes such as isoamylases and pullulanases constitute
another group of starch processing enzymes. They are mainly debranching
enzymes that cleave the-(16) glycosidic bonds. Transferases are also
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starch converting enzymes that cleave -(14) glycosidic bonds and transfer
a part of the donor to a glycosidic acceptor along with formation of new
glycosidic bond.

The production of glucose syrup from starch begins by dissolving starch

granules in the water by boiling or using jet cooker. The heated starch slurry is
passed into a hydrolysis reactor for enzymatic treatment. The degree of starch
sweetness depends on the degree of hydrolysis. Generally, the production of
maltose or glucose syrup from starch has three stages: gelatinization,
liquefaction and saccharification. In industries, high glucose syrup can be
converted to high fructose syrup using enzyme D-xylose-ketol isomerase.
Fructose, as you know is an isomer of glucose. It is twice as sweet as
glucose.

11.2.3 Enzymes in Baking Industry

Bread baking industries are now seeking an extensive usage of enzymes.

Starch modifying enzymes are playing a key role in baking and its different type
of applications. Flour components are hydrolyzed by different enzymes for
example, amylase hydrolyses starch, xylanases degrades xylan, proteases
degrade proteins and lipases hydrolyse the lipids. Oxidative enzymes like
glucose oxidase and lipoxygenase also act on different components of flour.
These enzymes affect the doughness or other properties of bread. Xylanase
increases the loaf volume and along with amylase gives softer morsel and
extended shelf life. Cell wall hydrolyzing enzymes such as cellulase, pectinase
and xylanase also play an important role in baking. Xylanases and glucanses
improve the elasticity of rye bread. Hemicellulases makes the cell wall more
soluble in wheat dough. Wheat flour includes about 1-1.5 % lipids. Non-polar
lipids such as triglycerides are bound with gluten. The addition of fungal lipase
hydrolyses the triglycerides which are bound to gluten and modify their
interactions in wheat flour. It leads to the formation of stable dough and
improved morsel structure. Glucose oxidase from Aspergilus niger and
Penicillium are known to oxidize alpha-D-glucose to gluconon-1,5-lactone. It
helps in improving the storage capability of flour.

Enzymes are used in the making of biscuits and nuts. The soft flour used in
making biscuits is different from those in bread making. It is basically a form of
dough with more elasticity in which gluten should not be too strong. Generally,
sodium bisulphate is used to produce the desired gluten but it is becoming
unpopular because of its potential health hazards. Sodium bisulphite has been
banned in many countries. Instead thermo stable proteases from Bacillus
subtilis are used nowadays to solve this problem by softening the gluten
without affecting the quality of dough.

SAQ 2
State whether following statements are True or False:

a) Isomerase is used to change glucose syrup into

166 fructose syrup. (True/False)
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b) Amylase and protease enzymes are used for laundering lot of
dirty clothes and fabrics. (True/False)

c) Proteases degrade lipids in the baking industry. (True/False)

d) The soft flour used in making biscuits is not different from

those in bread making. (True/False)

11.2.4 Enzymes in Fruit juice, Brewing and Wine

Industry

Do you take fruit juices? Do you anticipate the role of enzymes in fruit juices?
Fruit juice industry is rapidly increasing nowadays. The industry faces several
problems in maintaining the stability of fruit juices. The fruit juices turn turbid or
cloudy due to the presence of compounds like cellulose, hemicelluloses,
xylans and arabans. The problem is even more acute with citrus fruits. Pectic
compounds are complex colloidal acid polysaccharides constituting -(1—>4)
galacturonic acid residues with side chains consisting of xylose, galactose or
L-rhamnose or arabinose. When the tissue is grinded, pectin rich fruit gives
high viscosity to fruit juice. The usage of pectinases helps to lessen the
viscosity of fruit juice and increases the pressability of pulp. The yield of fruit
juice also increases significantly. Apple juice is filtered clear by the usage of
pectic enzymes. They help in processing of the fruit pulp as well as in fruit juice
extraction. Pectinase such as rhamnogalacturonase aids in the maceration of
apple tissue. Pectinases depolymerizes highly esterified pectin. Combination
of pectinesterases, polygalactouronases, polymethylgalactouronate lyases
improves the clarification of pear juice. Orange juice includes large amounts of
pectin esterase to take away methoxyl groups from pectin molecules. Mango
fruit is extensively used for making juice or squash. Mango nectar has 25-30 %
squash of pH 3.5 and titrable acidity of 0.2-0.4 %. Pectinases, hemicellulases
and cellulases enzymes assist in degrading mango pulp to make a plain, non-
gelatinous and clarified juice. Similarly, a mixture of enzymes like
pectinesterase, arabanase, hemicellulase, tannase and cellulases are used to
clarify the guava juice. Some of the bitter compounds present in citrus fruits
especially grapes are naringin, limonin and neohesperidin. The enzyme
naringinase from several species of microorganisms catalyzes the hydrolysis
of naringin to rhamnose and naringenin (a non-bitter derivative). Naringinase is
an enzyme complex constituting L-rhamnosidase and glucosidase and
therefore helps to remove rhamnose and glucose from naringin.

Beer is made by mixing barley malt with hot water in large vessel using
mashing. Maize, sorghum, rice and barley and other starchy cereals are also
added to the mash. The filtered mash is boiled and cooled and then transferred
to fermentation vessels and thereafter yeast is added in these vessels. After
fermentation, green beer is matured for additional filtration and bottling. The
enzymes used to convert cereals to beer are barley malt. This malted barley is
a major source of enzymes in the brewing process of wine industries. Acidic
pectinases from fungal sources such as Aspergillus niger are widely used in
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fruit juice and wine industries. Under optimal conditions, activation of many
enzymes such as -amylases, carboxypeptidases and -glucanase leads to
hydrolysis of starch materials in the seed reserve which is eventually used for
the beer production. Bacterial and fungal amylase enzymes are added for
saccharification. Pectinases and hemicellulases enzymes play a significant
role in the wine industry as they help in clarification, filtration and also improve
wine quality and stability.

11.2.5 Enzymes in Food Industry

Proteases enzyme are being used in food industry from number of years.
Papain from the leaves of unripe fruit helps in tenderizing the meat. Rennet
from the stomach of calves is traditionally used for cheese production.
Proteases aids in extraction of proteins from the dead animals for the
preparation of canned meats and soups. You must have heard of soya sauce
used extensively in fast food industry. Proteolysis in soya protein can increase
the value of their usage in the making of soya sauce. A partial proteolysis
augments whipping expansion of soya protein and its emulsifying properties.

Proteases help to recover proteins from animal tissues after butchering. The
bones from the animal are mashed with alkaline proteases and incubated at
65oC up to four hours. The meat slurry formed is used in canned meats and
soups.

The animal feed industry is a significant division of agro-business worth more

than 100 billion dollars. The major components of this industry include poultry,
pigs and ruminants, pet foods and fish farming. Majority of the food
constituents are not entirely digested by livestock. The application of enzymes
increases their digestibility. The enzymes can be added to the feed directly or
premixed with vitamins, minerals or other food additives. Hydrolases eliminates
the anti-nutritional factors present in the grains or vegetables. These enzymes
are known to increase the digestibility of animal feed as well as for enhancing
the nutritional value of feed. They also supplement animals with their own
digestive enzymes like amylases and -glucanases to hydrolyse non-starchy
polysaccharides such as barley. The non-starchy polysaccharides in cereals
results in poor feed conversion rate, slow weight gain and sticky droppings by
young chicks. Enzymes such as -glucanases and xylanases degrade these
polysaccharides improving their digestion as well as absorption to increase
weight gain and egg laying hens.

Cellulases and hemicellulases have great potential applications in the animal

feed industry. The increase milk yield and body weight by ruminants. These
enzymes hydrolyze plant cell wall polysaccharides since the fodder diet of
ruminants contains cellulose, hemicelluloses, pectin and lignin. Dairy cows
have shown a 5-20% increase in the production of milk when they are fed with
forage treated with fibrinolytic enzymes.

11.2.6 Enzyme in Other Industries

Textile, paper and pulp industry uses chemical reagents of diverse

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known to be carcinogenic. The dye wastewater is treated with ligninolytic
enzymes like lignin peroxidase and manganese peroxidase to degrade dyes,
including synthetic ones used in the industry. Thermostable enzymes like
cellulases, xylanases and proteases are also used in producing good quality
yarn or cotton. Laccases, polyphenol oxidases, lignin peroxidases, reductases,
methyl transferases and cytochrome oxygenases play a big and vital role in the
degradation of toxic xenobiotics. The material used in making fabrics contains
cellulosic fibres such as cotton, linen, viscose and lyocell. These materials
become ‘fuzz (short fibers sticking out from the surface)’ or fluffy. These
shortcomings in the cellulosic fibres can be eliminated using cellulases in
process called as bio-polishing. This process is carried out during textile wet
processing stage that includes desizing, bleaching, dyeing and finishing. The
enzyme cellulases act on small fibres and polish them. They also help in
smoothening, increasing the moisture absorbance and giving glossy
appearance to the fabrics. Similarly, acid cellulases have demonstrated to be
successful in treating lyocell garments by giving them a soft feel look and
preventing piling. Fungal -amylases are also used in desizing woven fabrics
without harming the yarn.

Paper industry involves separation and degradation of lignin in the wood pulp.
The new environment issues stresses the need to replace conventional
pollution emanating chlorine-based delignification/bleaching processes. The
treatment of wood pulp with ligninolytic enzymes addresses this apprehension
by providing gentle and cleaner strategies of delignification. The usage of
enzymes in bio bleaching systems is also providing additional help to the paper
industry. Another category of enzymes like lacasses can be used in targeted
modification of wood fibres. Enzymes such as cellulases and hemicellulases
are also used in paper and pulp industry for several purposes. Mechanical
pulping in refining and grinding of the woody raw material provide pulps with
elevated content of fines, bulk and stiffness but the processes involve high-
energy consumption. Bio-mechanical pulping results in substantial energy
savings. In bio-bleaching processes, pretreatment of lignocellulosic materials
with microorganisms seems promising because microorganisms produce
hemicellulolytic enzymes (xylanases) that limits the hydrolysis of
hemicelluloses in pulp to increase the extractability of lignin.

The milk and dairy products are one of the oldest industries which remain
applicable till today. The world’s adult population is around 75 % today. The
probable market of milk and dairy products industry is still unrecognized. The
commercial production and development of enzyme galactosidase (lactase)
from microbial sources has made dairy industry significantly practicable. Milk
and milk products, cheese, condensed milk and ice cream are several
products daily used in dairy industry. If you recall from your unit on
carbohydrates, lactose is a disaccharide made of D-glucose and D-galactose.
Lactase from microorganisms hydrolyzes lactose. It can be produced
commercially from dairy yeast Kluyeromyces fragilis at pH optimum (6.5-7.2)
with milk treatment. Yeast -galactosidases are used for lactose hydrolysis in
milk and sweet whey. Milk acidification is required for release of whey from the
cheese. Lactic acid bacteria such as Lactobacillus sp convert lactose to lactic
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acid. Several varieties of cheese can be made using starter ‘a culture inoculum
containing lactic acid bacteria’. Casein fraction of milk can be coagulated to
form a gel by lowering the milk pH and adding up rennet ‘mixture containing
specific proteolytic enzymes’. Chymosin enzyme can coagulate about 70 % of
the cheese.

SAQ 3
Do as Directed

a) Pectinases polymerize/depolymerize highly esterified

pectin. (Choose one option)

b) Name two enzymes used in producing good quality yarn or cotton.

c) ................... and .....................enzymes play a significant role in

clarification and improvement of wine quality and stability. (Fill in the blank)

d) Hydrolases eliminates the antinutritional/ nutritional factors

present in the grains or vegetables. (Choose one option).

11.2.7 Enzymes in Medicine/Diagnostic Enzymes

In your visit to the hospital, you may come across biochemical tests prescribed
by the physician for the determination of disease or its progression. Many
enzymes present in plasma or serum are routinely tested in clinical labs as
these enzymes are of diagnostic value. Each plasma enzyme has a normal
concentration range due to the balance that is set up between its rate of arrival
and its rate of removal in the blood stream. These enzymes have a normal
range of activity. If a particular tissue is affected by disease or gets damaged,
some of enzymes may leak into the bloodstream at an increased rate and will
be found in the plasma in elevated amounts. The detection and measurement
of enzymes using biochemical methods helps in detection of disease via their
correlation with the symptoms, progression of disease and even cure from the
disease. Table 11.2 enlists several diagnostic enzymes and their potential
uses.
Table 11.2: Diagnostic enzymes

Enzyme Abbreviations Use

Serum glutamic oxaloacetic SGOT/AST Heart attack, Liver

transaminases/ Aspartate dysfunction
transaminase

Serum glutamate-pyruvate SGPT/ALT Liver dysfunction

transaminases/ Alanine
transaminase

Creatine Kinase CK Brain and muscle

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Urease Renal function test

Cholesterol Oxidase Hypercholesterolemia,

Heart attack

Lactate Dehydrogenase LDH Myocardial Infraction

Alkaline Phosphatase ALP Bile and Liver diseases

Acid Phosphatase ACP Prostate Cancer

Glucose Oxidase Diabetes Mellitus,

Hyperglycemia

11.2.7.1 Enzyme Therapy

In the enzyme therapy, enzymes are added to change the existing abnormal or
diseased state owing to the lack or short supply of same enzyme in the body.
Enzymes are given most likely in an immobilized manner but biodegradable
form. Enzymes such as L-asparaginase are used in the treatment of
Leukemia. Streptokinase enzyme helps to get rid of blood clots in the blood
vessels and play an important role in curing heart attacks. Enzyme therapy
has been involved in the degradation of necrotic tissue by usage of proteolytic
enzymes such as trypsin or chymotrypsin.

11.2.7.2 Enzymes and Inborn errors of Metabolism

Inborn errors of metabolism occur due to genetic mutations and are

characterized by the loss of enzyme activity. More than 400 errors are known
but they occur rarely, for example phenylketonuria, 1 in 12,000 births;
cystinuria, 1 in 14,000; galactosemia, 1 in 57,000. These errors are identified
by enzyme assays or the presence or absence of particular metabolite in
blood, tissue, urine or other biological samples. The advent of recombinant
technology has led to the development of highly accurate and sensitive
diagnostic procedure for determining enzyme activities. It is feasible to obtain a
pre-natal diagnosis by carrying out enzyme assays in the amniotic fluid. Some
examples of inborn errors of metabolism due to enzyme deficiencies are given
in Table 11.3:

Table 11.3: Enzyme deficiency and inborn errors of metabolism

Inborn Error Enzyme Deficiency

Alkaptonuria Homogentisate1,2 –dioxygenase
Phenylketonuria Phenylalanine 4 monooxygenase
Galactosemia Galactose 1-phosphate uridyltransferase
Glycogen storage disease type V Glycogen phosphorylase
Tay-Sachs -N-Acetyl-D-hexosaminidase

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Treatment of inborn errors of metabolism usually involves reduction in the
intake of substances that cannot be metabolized normally or supplementation
of special diets to the patients. However, this kind of treatment is ineffective
because of endogenous synthesis of metabolites in question or due to
blockage of particular metabolic pathway vital for the body function. A
successful treatment is possible only when normal enzyme could be
administered at the appropriate site for example urokinase plasminogen
activator (uPA) extracted from human urine can be infused in the patients
suffering from pulmonary embolism. L-Asparaginase is also used in the
treatment of leukemia.

SAQ 4
Answer the following

Name the enzymes used for detection of liver disorders and state their role.

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11.2.8 Enzyme Electrodes

Immobilization technology has led to the usage of enzymes as electrodes

which are applied in principle by autoanalyzers to carry out biochemical as well
as pathological estimations in various biological samples such as plasma,
serum, urine and cerebrospinal fluid.
D-Glucose oxidase enzyme is immobilized on a polyacrylamide gel. It
catalyzes the following reaction:
D-Glucose + O2 D-Glucono-1,5-lactone + H2O2
The determination of the depletion of oxygen by the oxygen electrode helps in
measurement of glucose levels. Table 11.4 enlists enzymes which can be
used as electrodes along with their potential usage.

Table 11.4: Enzymes as electrodes

Immobilized Enzyme Sensing Electrode Compounds

Analyzed

Urease Carbon dioxide Urea

Lactate Dehydrogenase Platinum Lactic acid

Alcohol Dehydrogenase Platinum Ethanol

Penicillinase pH Penicillin
Cholesterol oxidase Oxygen Cholesterol
Glutaminase Cation Glutamine
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Immobilization of enzymes has led to the development of new analytical
devices called biosensors. These sensor devices are used to measure the
substances and parameters of biological interest. These devices convert the
biological response to an electronic signal which is amplified, processed and
displayed. Biosensors have now found an extensive usage in the analytical
chemistry.

11.3 SUMMARY
1) Practical interest in enzyme processes has led to the development of
enzyme industry which runs in hundreds of millions.

2) Enzymes are used commercially for the reason of their catalytic abilities.
They are beneficial because of low costs and their usage in minimal
quantities.

3) The applications of enzymes are huge and varied for example in food,
starch, detergent and baking industry.

4) In food industry, enzymes such as amylases, lactases and cellulases

break composite sugars to simple ones. Alkaline proteases from microbial
origin are extensively used in the detergent industry.

5) Cell wall hydrolyzing enzymes such as cellulase, pectinase and xylanase

play a significant role in the baking industry.

6) Enzymes such as endoamylases, exoamylases, debranching enzymes

and transferases are daily used in starch industry.

7) Pectinase act on hard pectin which is used in fruit juice preparation.

These enzymes help in making juice less viscous.

8) The scope of using enzymes for medical diagnostics is also widening

rapidly. More and more enzyme assays are now being performed on
autoanalyzers. Several enzymes give clinical distinguishment between
different diseases. Inborn errors of metabolism are also treated using
enzyme therapy.

11.4 TERMINAL QUESTIONS

1) What makes enzyme catalyzed reactions more suitable for industrial
applications?

2) Describe the role of enzymes in the starch industry.

3) Describe how biological washing powders work?

4) Discuss the disadvantages in the usage of enzymes.

5) Prepare a list of diagnostic enzymes and their uses in the detection of

diseases.

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11.5 ANSWERS
Self-Assessment Questions

1) a) Microbes can be easily grown in large numbers in bioreactors at

nominal cost using simple nutrients. They can also withstand
extreme pH, temperature or dissolved oxygen like conditions. So,
these are ideal choice as enzyme source.

b) Genetic Engineering, Cloning, Genetic Recombination

c) Extracellular enzyme, Ease of isolation and purification

2) a) True, b-True, c) False, d) False

3) a) depolymerizes, b) Cellulases, xylanases, proteases, c) pectinases

and hemicellulases, d) anti-nutritional.
4) Enzymes used for the detection of liver disorders are:
a) Alkaline Phosphatase: This enzyme is found in high concentrations in
liver and is tested to determine liver functioning and to detect liver
lesions that may cause biliary obstruction such as tumors or
abscesses.
b) Alanine transaminase (ALT): This enzyme is released in the
bloodstream after acute liver cell damage.
c) Aspartate transaminase (AST): This enzyme is released in the
bloodstream after acute liver cell damage.
d) Gamma-glutamyl transpeptidase test: This enzyme test is used to
assess liver function, information about liver disease and to detect
alcohol ingestion.
e) Lactic Dehydrogenase: It detects tissue damage and assist in
diagnosis of liver disease.
f) 5’-Nucleotidase: This enzyme is elevated in persons with liver
diseases especially those with cholestasis (disruption of, or
obstruction in the flow of bile).

Terminal Answers:

1) Industrialists were the first to discover the potential of enzymes as they

realized if the reactions are needed to be speeded up, production
processes could be performed in only a fraction of the normal time or at
low temperature or pressure or using economical starting material. As a
result, enzymes are finding widespread usage from bio-bleaching of paper
to efficient recovery of oil and gas. Usage of enzyme is considered to be
safe, cost-effective or environment friendly. Food, detergent or beverage
industries are heavily dependent on enzymes as well as the role of
enzymes in bio-analysis have become extensive nowadays.
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Unit 11 Applications of Enzymes
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2) Refer to section 11.2.2

3) Biological washing powders contain lipases and proteases which breakdown

the proteins and fats in stains. For details refer to section 11.2.1

4) Enzymes are highly sensitive to the surrounding environment which

makes them hard to use. They get denatured due to small changes in pH
and temperature. They are also highly susceptible to poisons. Isolation
and purification of enzymes involve huge costs and complex separation
techniques. Enzymes substrate mixture must be uncontaminated as other
substances in the reaction mixture may affect the reaction. All these above
mentioned disadvantages make usage of enzymes very hard as the
conditions involving them must be tightly controlled.

5) Refer to Table 11.2

11.6 SUGGESTED READINGS

1) David L. Nelson and Michael M. Cox: Lehninger Principles of Biochemistry
6th Ed., W.H. Freeman.

2) Robert K. Murray, Daryl K. Granner, Victor W. Rodwell Harper’s Illustrated

Biochemistry, 27th edition. 2006, McGraw-Hill.

3) Donald J Voet and Judith G. Voet:Principles of Biochemistry4th ed., John

Wiley and Sons, Inc, USA.

4) Eric E Conn, Paul K Stumpf: Outlines of Biochemistry, Jophn Wiley and

Sons, Inc, USA.

5) S. Shanmugan and T. Sathishkumar: Enzyme Technology, I K International

Publishing House Pvt Ltd, New Delhi.

6) Nicholas C Price and Lewis Stevens: Fundamentals of Enzymology,

Oxford University Press, Oxford, New York, USA.

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