‫ﺑﺴﻢ ﷲ اﻟﺮﺣﻤﻦ اﻟﺮﺣﯿﻢ‬

Industrial Enzymes

Bioc. 231

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 Industrial enzymes
• Industrial enzyme: an enzyme that is used to facilitate
industrial processes or the production of industrial
products.
• Over 500 products covering 50 applications, from
detergents to beer making, utilize enzymes produced
through large-scale fermentation of microorganisms.
• As both rational and random methods of enzyme
improvement have evolved over the past 20 years,
enzymes are being developed to play an ever-
expanding role as catalysts in industrial processes.

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 Industrial enzymes
• The estimated worldwide industrial enzyme market in 2000
was valued at approximately $1.5 billion and is traditionally
divided into three segments.
1. The largest, at 65% of sales, is that of technical enzymes
and includes enzymes used in the detergent, starch,
textile, leather, pulp and paper, and personal care
industries.
2. Food enzymes, the second largest segment at 25% of the
market, includes enzymes employed in the dairy, brewing,
wine and juice, fats and oils, and baking industries.
3. Finally, feed enzymes, comprising enzymes used in animal
feeds, contributes approximately 10% of the market.

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 Industrial enzyme
• Although it has dated back to the ancient times when
enzymes were used in baking, brewing, cheese making,
etc., they were used either as spontaneously growing
microorganisms or as added preparations such as
calves’ rumen or papaya fruit.
• Only during the past few decades, the development in
recombinant DNA technology and advanced
bioprocesses made it possible to produce enzymes as
purified, well-characterized preparations on a large
scale, allowing the wide application of enzymes in
various industrial products and processes, such as
chemical, detergent, textile, food, animal-feed, leather,
and pulp and paper industries.
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• Kirk O, Borchert T and Fuglsang C. Industrial enzyme applications. Current Opinion in Biotechnology 2002, 13:345–351.
 Proteases
• Proteases are a large group of enzymes that catalyze the
hydrolysis of peptide bonds in proteins and polypeptides.
• Proteases may be classified into 3 groups by the optimal pH
in which they are active:
1. Acidic proteases: have pH optima in the range of 2.0–5.0
and are mainly fungal in origin.
2. Neutral proteases: have pH optima around 7.0. They
aremainly of plant origin and some bacteria and fungi also
produce neutral proteases.
3. Alkaline (basic) proteases: have pH optima in the range of
8.0–11.0. Some of the important alkaline proteases are
those from Bacillus and Streptomyces species.

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 Proteases
§ There are thousands of different protease molecules that
have been isolated and characterized. Among them,
several hundred proteases are commercially relevant,
and have been used in laundry and dishwashing
detergents, food processing, animal-feed additives,
leather processing, waste treatment, pharmacology, and
drug manufacture.
§ The main industrial application of proteases is their use
as detergent additives to remove protein deposits and
stains, and the major player is subtilisins.
• Subtilisin are used in the detergents for dish washers and
all types of powder and liquid laundry detergents as well
as in laundry bleach additives. The subtilisin
concentration in detergent and cleaning products is very
low and depends on the type of product, typically
ranging between 0.007% and 0.1%.
• Subtilisins (<10%) are also used in technical applications
such as protein hydrolysate production, leather
treatment, and in the textile and cosmetics industry.
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 Proteases
• Proteases are a powerful tool for modifying the
properties of food proteins, and they are widely
used in the production of value-added food
ingredients and food processing for improving
the functional, nutritional, and flavor properties
of proteins.
• The function of proteases is to catalyze the
hydrolysis of proteins, which has been exploited
for the production of protein hydrolysates of high
nutritional value from casein, whey, soy protein,
and fish meat.

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 Lipases
• Lipases are a class of hydrolases that catalyze the
hydrolysis of triglycerides to glycerol and free fatty
acids.
• Lipases are widely present in bacteria, fungi, plants,
and animals.
• Lipases are the enzymes of choice for potential
applications in numerous industrial processes
including areas such as oils and fats, detergents,
baking, cheese making, hard-surface cleaning, as
well as leather and paper processing.
• Nowadays, heavy-duty powder detergents and
dishwasher detergents usually contain one or more
enzymes including lipase, which can reduce the
environmental impact of the detergent products.
Lipases are used in detergent formulations for
efficient removal of lipid stains, allowing better
washing performances and energy savings.
• .
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 Lipases

• Biodiesel is an alternative diesel fuel consisting of

short-chain alkyl (methyl or ethyl) esters, typically
made by trans-esterification of vegetable oils or
animal fats. The conventional method for producing
biodiesel, involving acid or base catalysts to form
fatty acid alkyl esters, results in high downstream-
processing costs and serious environmental
problems. Thus, lipase-catalyzed transesterification
presents an excellent alternative for biodiesel
production.

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 Amylases
§ Amylases are glycoside hydrolases that act on α-1,4-
glycosidic bonds and break down starch into sugars.
§ Amylases of microbial origin are divided into exo-and endo-
acting enzymes.
• Exo-amylases include glucoamylases and β-amylases.
- Glucoamylases (EC 3.2.1.3) catalyze the hydrolysis of α-1,4
and α-1,6 glucosidic linkages with lower rate for α-1,6
cleavage to release β-D-glucose from the non reducing
ends of starch and related poly-and oligosaccharides.
- β-Amylases (EC 3.2.1.2) cleave the α-1,4-glycosidic bonds
in starch from the non reducing ends to give maltose.
• Endo-amylases (α-amylases, EC 3.2.1.1) hydrolyze internal
α-1,4 bonds and bypass α-1,6 linkages in amylopectin and
glycogen at random in an endo-fashion producing malto-
oligosaccharides of varying chain lengths.
§ The amylases of microorganisms have a broad spectrum of
industrial applications as they are more stable than those
of plant and animal origin.
§ Although α-amylase has been derived from various species,
enzymes from fungal and bacterial sources have dominated
applications in industrial sectors

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 Amylases
§ α-Amylase is one of the most popular and important forms of industrial
amylases, and has potential application in a wide number of industrial
processes such as food, fermentation, textile, paper, detergent, and
pharmaceutical industries.
§ α-Amylases also play a dominant role in carbohydrate metabolism and have
been isolated from diversified sources including plants, animals, and
microbes.
• α-Amylases are extensively employed in the bakery industry. During baking,
the α-amylase added in the dough of bread catalyzes the degradation of
starch in the flour into smaller dextrins, which are subsequently fermented
by the yeast to generate CO2. This process results in improvements in the
volume and texture of the product.
• In addition, α-amylases in bread baking also generate additional sugar in the
dough to improve the taste, crust color, and toasting qualities of the bread.
• Furthermore, α-amylases have an antistaling effect in bread baking and
extend the softness retention time of baked goods, thus increasing the shelf
life of these products. 12
 Pullulanases
§ Starches are one of the most widely used polysaccharides,
predominantly α-1,4-linked D-glucopyranoside polymers with
different degree of α-1,6 branches.
§ In addition to the enzymes that hydrolyze the α-1,4 linkages,
complete starch hydrolysis also requires α-1,6-glycosidic cleaving
enzymes including pullulanases.

§ Pullulanases (EC 3.2.1.41) are widely distributed among animals,

plants, fungi, and bacteria.
• Type I pullulanases specifically attack α-1,6-glycosidic linkages in
branched oligosaccharides such as starch, amylopectin, and
glycogen, forming maltodextrins linked by α-1,4-glycosidic linkages.
• Type I pullulanases and isoamylase (EC 3.2.1.68) are called as
debranching enzymes that exclusively hydrolyze α,1-6 glycosidic
bonds.

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 Pectinases
• Pectinases or pectinolytic enzymes are a
heterogeneous group of related enzymes that
hydrolyze the pectic substances.
• Pectic substances are high-molecular-weight,
negatively charged, acidic complex glycosidic
macromolecules (polysaccharides) that are mostly
present in the plant. The primary backbone of pectic
substances consists of α-D-galacturonate units linked
α-1,4, with 2–4% of L-rhamnose units linked β-1,2 and
β-1,4 to the galacturonate units.
• The largest industrial application of pectinases is in
fruit juice extraction and clarification. A mixture of
pectinases and amylases is used to clarify fruit juices,
in which suspended matter is removed to give
sparkling clear juices (free of haze).

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 Laccases
• Laccases are enzymes that oxidize a large variety of organic substrates, as well as some inorganic
ions.
• The textile industry :degrading dyes of diverse chemical structure. Laccases are also used in textile
bleaching.
• Polycyclic aromatic hydrocarbons together with other xenobiotics are a major source of
contamination in soil; therefore, their degradation is of great importance for the environment.
Laccases can be used to degrade such compounds, including those arising from natural oil
deposits and utilization of fossil fuels.
• Laccases can be applied in different aspects of the food industry such as bioremediation, beverage
processing, ascorbic acid determination, sugar beet pectin gelation, and baking.

• A laccase from the white-rot fungus Trametes hirsuta increased the maximum resistance of dough
and decreased the dough extensibility in both flour and gluten dough. It is due to the ability of
laccase to cross-link biopolymers

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 Transglutaminases
§ Transglutaminases are a family of enzymes
(EC 2.3.2.13) that catalyze the formation of a
covalent bond between a free amine group
of protein-or peptide-bound lysine (acyl
acceptors) and the gamma-carboxamide
group of protein-or peptide-bound
glutamine (acyl donors). This results in the
modification of proteins through either intra-
or intermolecular cross-linking, thus
improving the end use of the protein.
§ Transglutaminases are widely present in
animal tissues and body fluids, plants, fishes,
and microorganisms.
§ Transglutaminases can be used to modify the
functional properties of food proteins by
catalyzing the cross-linking of a number of
proteins, such as whey proteins, soya
proteins, gluten, meet, and fish protein .

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 Transglutaminases
§ The modification of food proteins by transglutaminase showed several
advantages:
• It helps to protect lysine in food proteins from various chemical reactions, thus
increasing the shelf life of food.
• Food proteins of higher nutritive value can be produced through cross-linking
of different proteins containing complementary limiting essential amino acids.
• Transglutaminases can encapsulate lipids and/or lipid-soluble materials, form
heat-and water-resistant films, avoid heat treatment for gelation, improve
elasticity and water-holding capacity, and modify solubility and functional
properties.
• Transglutaminases have also been reported to improve the quality of wool as
in felting, whitening, handling, and shrinking.

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 Phytases
• Phytases (myo-inositol hexakisphosphate phosphohydrolase, EC 3.1.3.8) catalyze the hydrolysis of phytate (myo-
inositol (1,2,3,4,5,6)hexakisphosphate) to the inorganic phosphate and less-phosphorylated myo-inositol
derivatives.
• They have been identified in plants, microorganisms, and in some animal tissues, and represent a subgroup of
phosphatases that initiate the stepwise dephosphorylation of phytate, the most abundant inositol phosphate in
nature.
• Phytases are classified into 3-phytases (EC 3.1.3.8), 6-phytases (EC 3.1.3.26), and 5-phytases (EC 3.1.3.72).
• Depending on their pH optima, phytases have been divided into acid and alkaline phytases.
• Phytases have been mainly used as animal-feed additive in diets largely for swine and poultry, and to some
extent for fish.
• Most commercially available phytases are of fungal or of bacterial origin genes, but expressed for production
purposes in yeasts.

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 Phytases
• Phytases are a cost-effective alternative to ensure animals to obtain adequate
available phosphorus from the plant-based diets, because they can turn the plant
phytate into a very valuable resource of phosphorus by improving its bioavailability
for animal nutrition.
• Use of the enzyme phytase as an animal feed supplement increases the nutritional
value of the feed by releasing phosphate and bound metals from the phytate. In
addition, it reduces the release of free phosphorus in animal waste by as much as
30%.
• Effects of supplemental microbial phytases on digestibility of starch, protein, and
amino acids have also been reported, but the data are inconsistent.
• Phytases catalyze the degradation of phytate, resulting in the release of minerals.
Thus, phytases have also been found increasingly used in processing and
manufacturing of food for human consumption.
• Furthermore, phytases may find application in the production of functional foods
or food supplements with health benefits.
• Phytase-catalyzed phytate degradation is also helpful for bread making, production
of plant protein isolates, corn wet milling, and the fractionation of cereal bran.
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