AMYLASE ECONOMIC AND APPLICATION VALUE
Transcript of AMYLASE ECONOMIC AND APPLICATION VALUE
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Α AMYLASE ECONOMIC AND APPLICATION VALUE
Nagib A. Elmarzugi1,2,3, Hesham A. El Enshasy1, Mariani AbdulHamid1, Rosnani
Hasham1, Azila Aziz1, Elsayed A. Elsayed4, Nor Zalina Othman1, Mohamed Salama5,*
1Institute of Bioproduct Development (IBD), Universiti Teknologi Malaysia (UTM), 81310
Skudai, Johor, Malaysia. 2Dept. of Industrial Pharmacy, Faculty of Pharmacy, Tripoli University, Tripoli, Libya. 3BioNano Integration Research Group, Biotechnology Research Center, NASR, Libya.
4Bioproducts Research Chair, Zoology Department, Faculty of Science, King Saud
University (KSU), Kingdom of Saudi Arabia. 5Faculty of Pharmacy, Universiti Teknologi Mara, Malaysia
ABSTRACT
The years of the research in biotechnology has boosted the importance
of enzymes in industry, as well as advanced characteristics of using
bioprocess rather than chemical synthesis in their preparation.
Enzymes are important in biological processes because they accelerate
specific biochemical reactions to produce a useful product or effect.
Enzymes are widely used in industrial processes due to their low cost,
large productivity and vast availability. Amylases are one of the most
important enzymes in present-day biotechnology. Amylases contribute
to world enzyme market about 25-30%. The estimated value of world
market of amylase industry is about US$ 2.7 billion at 2012 and
estimated to increase by 4% annually. α-amylase has variable wide
applications. Textile de-sizing based on thermo-stable bacterial
amylase covers about 12% of amylase market. α-amylase uses a very
small quantity of bio-catalysis in bio-washing products instead of very high quantity of
chemical detergents. In food production, enzymes are preferred to alternate the chemical-
based technology. α-amylase is strongly integrated with medical and pharmaceutical
applications. These applications include therapeutic and diagnostic tools for managing a quite
number of diseases ranging from ordinary problems to gene therapy by correcting the enzyme
deficiency.
World Journal of Pharmaceutical ReseaRch
7105 – 2277ISSN Article Review . 4906-4890Volume 3, Issue 3,
Article Received on 12 March 2014,
Revised on 05 April 2014, Accepted on 28 April 2014
*Correspondence for
Author
Prof. Dr.Mohamed Salama
Department of Pharmaceutics,
Faculty of Pharmacy,
Universiti Teknologi Mara,
Level 11, FF1 Building, 42300
UiTM Puncak Alam Campus,
Selangor, Malaysia.
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Key Words: Enzyme, α-amylase, textile de-sizing, bio-washing, Starch.
INTRODUCTION
Enzymes play a key role in almost all biological processes, accelerating a variety of
metabolic reactions as well as controlling the energy for genetic information pathways.
Enzymes are biological catalysts responsible for supporting almost all of the chemical
reactions that maintain general homeostasis; because of their role in maintaining life
processes (Kamerlin and Warshel 2010). They are tools of nature and using natures’ own
technology in modern industries, associated with many advantages, compared to chemicals,
enzymes are specific in their action and work at mild conditions (M., S. et al. 1996). The
enzyme is also called Biocatalyst in order to catalyse any reaction. Biocatalysts are vitally
important in the industrial process to be fully exploited. Biocatalysts can be either intact cells
or isolated enzymes (Pisliakov, Cao et al. 2009). The special properties of biocatalysts
include conversion of substrates into the desired products both in single reactions and in
complex cell systems, synthesis of products under mild reaction conditions, which
approximate ambient conditions with regard to pressure, temperature and pH value.
Therefore, using of enzymes in bioprocess industry could offer a number of advantages such
as thirty percent saving in raw materials during production process, and up to thirty percent
energy savings with mechanical process in production (Bonnet and Millet 1996). Enzymes
are proteins, composed of hundreds of amino acids, which are produced by living organisms.
They are responsible for number of reaction and biological activities in plant, animal, human
beings and microorganisms. They are found in the human digestive system to break down
carbohydrates (sugars), fats or proteins present in food. The smaller pieces can be absorbed
into the blood stream; each 10-25% decrease in chlorine chemicals by substituting Xylanase
before the first bleaching step. Moreover, the use of this method in the European paper
industry could cut annual CO2 emissions by 155.000 to 270.000 tons. Therefore, reduction in
the use of chemicals, e.g. chlorine, EDTA and peroxides, by enzymatic deinking of recycled
paper by lipase, cellulases, xylanases or pectinases. The economical paper production by
efficient use of recycling paper and more protection of the environment by reducing the
volume of waste paper and the demand for wood (Namvar-Mahboub and Pakizeh 2012).
Enzymes are catalysts and most of them are protein and few ribonucleoprotein enzymes have
been discovered and, for some of these, the catalytic activity is in the RNA part rather than
the protein part (C., Q. et al. 2011). Enzymes bind temporarily to one or more of the
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reactants of the reaction they catalyze. In doing so, they lower the amount of activation
energy needed and thus speed up the reaction, which is represented in figure 1.
Figure1. Effect of activation energy on chemical reaction.
History of enzymes industry
The history of modern enzyme technology could be traced to the beginning of 1874 when
Hansen manufactured Chymosin from the stomach of calves for manufacturing of cheese.
This was the first enzyme preparation of relatively high purity used for industrial purposes.
Jokichi Takamine was the first person to manufacture an enzyme from a microbial source
when he manufactured taka diastase from Aspergillus as a digestive enzyme in 1894. The
method of fermentation suggested by Takamin, the "surface culture" is still actively used in
the production of various enzymes. Bacterial amylase derived from Bacillus Subtilis was used
for desizing, the first time by Boidin and Effront as early as 1917. In 1950, fungal amylase
was used in the manufacture of specific types of syrup, those containing a range of sugars,
which could not be produced by conventional acid hydrolysis. The real turning point was
reached early in 1960, when an enzyme glucoamylase was launched for the first time, which
could completely break down starch into glucose. The process was further improved by the
introduction of a new technique used for the enzymatic pre-treatment of starch by using a
heat stable alpha amylase (Johnson 2013).
Enzyme cofactors
Numerous enzymes require the presence of an additional, non-protein and co-factor. Many of
these are metal ions such as Zn2+ that act as co-factor for carbonic anhydrase, Cu2+, Mn2+, K+,
and Na+. Some cofactors are small organic molecules called coenzymes. The B vitamins
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[thiamine (B1), riboflavin (B2) and nicotinamide] are precursors of coenzymes (Sennett,
Kadirvelraj et al. 2012).
In the last few years the time for molecular biological optimization of enzymes has decreased
by a factor of 10-100, while at the same time the stability and productivity of selected
enzymes has unmistakably risen. In the past ten years, the market volume for enzymes rose
by approximately 50 %. A US study made in 2004 anticipates annual growth of the US-
American enzyme market of 6 % and a market volume of US$ 1.9 billion by 2008, the
percentage of total applications of enzymes by market sectors (Flaschel E, Bott M et al.
2006).
Table1. Distribution of the worldwide application of technical enzymes by market
sectors (Flaschel E, Bott M et al. 2006)
Market sector % share of total application Detergent 40 Cheese ripening and aroma 12 Flour and baking products 10 Leather processing 10 Glucose isomerization 7 Fruit utilization and wine 7 Starch degradation 5 Brewery (not in Germany) 5 Silage and feed 2 Pulp and paper, textile 2
Structure
According to computer modular simulation studies, the protein engineering of amylase
enzyme found in human saliva and pancreas. This enzyme can digest starch molecules and
break them down to sugar molecules. Enzyme is made of a sequence of amino acids folded
into a unique three-dimensional structure that determines the function of the enzyme. Even
the slightest change in the sequence of the amino acids can alter the shape and function of the
enzyme.Each enzyme consists of several hundreds of amino acids located in such a delicate
three-dimensional structure as in Figure 2 and 3. This structure determines the properties of
the enzyme such as reactivity, stability and specificity. Enzyme are essential for all
metabolic processes, but are distinguishable from other proteins because they are known as
biological catalysts or substances, which speed up reaction without get used up themselves.
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Figure 2. Salivary alpha-amylase Figure 3. Pancreatic alpha-amylase
Production of enzymes
Enzyme molecules are too complex to synthesize by purely chemical means. The problem is
that enzyme produced by microorganisms often expressed in tiny amount mixed up with
many other enzyme and proteins. These microorganisms can also be very difficult to cultivate
under industrial condition, and they may create undesirable by-product. Modern industrial
cultivation of enzymes begins with fermentation of a vial of dried or frozen microorganisms
called a production strain. This production strain is selected to produce large amounts of the
enzymes of interest. The production strain is first cultivated in a small flask containing
nutrients and a gar, the flask placed in an incubator, which provides the optimal temperature
for the previously frozen or dried cells to germinate. Once the flask is ready, the cells are
transferred to a seed fermented, which is a large tank containing previously sterilized raw
materials and water, known as the medium, seed fermentation allows the cell to reproduce
and adapt to the environment and nutrients that they will encounter later on. The cells are
then transferred to a larger tank, the main fermented, where temperature, pH and dissolved
oxygen are carefully controlled to optimize enzyme production, and additional nutrients may
be added to enhance productivity. Therefore, when main fermentation is complete the
mixture of cells, nutrient and enzyme, referred to as the broth, is ready for filtration and
purification.
Amylase Enzyme
Microbial enzymes are widely used in industrial processes due to their low cost, large
productivity, chemical stability, environmental protection, plasticity and vast availability (N.
Sajitha, V. Vasanthabharathi et al. 2011, Yasser Bakri, Hassan Ammouneh et al. 2012).
Bacillus species such as Bacillus subtilis, Bacillus amyloliquefaciens and Bacillus
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licheniformis are used as bacterial workhorses in industrial microbial cultivations for the
production of a variety of enzymes as well as fine biochemicals for decades. A large quantity
(20-25g/l) of extracellular enzymes has been produced and secreted by the various Bacillus
strains which have placed them among the most significant industrial enzyme producers. The
estimated value of world market is presently about US$ 2.7 billion and is estimated to
increase by 4% annually through 2012. Detergents (37%), textiles (12%), starch (11%),
baking (8%) and animal feed (6%) are the main industries, which use about 75% of
industrially produced enzymes (Nik C. Barbet, Ulrich Schneider et al. 1996, C. Meintanis,
K.I. Chalkou et al. 2008). Amylases are among the most important enzymes having great
significance for biotechnology, constituting a class of industrial enzymes having
approximately 25-30% of the world enzyme market (Deb, Talukdar et al. 2013). Initially the
term amylase was used originally to designate an extracellular enzymes capable of
hydrolyzing α-1,4-glucosidic linkages in polysaccharides containing three or more 1,4-α-
linked glucose units. The enzyme acts on starches, glycogen and oligosaccharides in a
random manner, liberating reducing groups. These enzymes are found in prokaryote as well
as in eukaryotic organisms. They are widely distributed in microbial, plant and animal
kingdoms (H., S. et al. 2013). Bacterial growth requires a source of energy in order to
assemble the various constituents comprising the cell. The energy supply is derived from the
controlled breakdown of various organic substrates present in the external environment, such
as polysaccharides, lipids, and proteins. Some members of the genus Bacillus are able to
break down starch and utilize it as an energy source. Organisms unable to produce amylase
cannot utilize starch as an energy source (Rothstein, Devlin et al. 1986, Pretorius I, Laing E et
al. 1988). Amylases are classified based on how they break down starch molecules into: I- α-
amylase (alpha-amylase) which reduces the viscosity of starch by breaking down the bonds at
random, therefore producing varied sized chains of glucose. II- ß-amylase (Beta-amylase)
which breaks the glucose-glucose bonds down by removing two glucose units at a time,
thereby producing maltose. III- Amyloglucosidase (AMG), which breaks successive, bonds
from the non reducing end of the straight chain, producing glucose (Carugo, Lu et al. 2001).
The Amylase enzyme has a shape that allows it to wrap around starch (substrate) and cut it up
into individual glucose units (Pandey A, Nigam P et al. 2000). α-amylase is an endo acting
enzyme, catalyzing the random hydrolysis of internal α1 to 4 glycosidic linkages present in
the starch substrate. These enzymes are incapable of hydrolysing α1 to 6 glycosidic linkages
present at branch points of amylopectin chains. One exception to this is the α-amylase
produced by thermoactinomyces vulgaris, which can hydrolyse both α1 to 6 and α1 to 4
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glycosidic linkages (Vihinen and Mantsiila 1989). α-amylase has been derived from several
fungi, yeasts, bacteria and actinomycetes, however, enzymes from fungal and bacterial
sources have dominated applications in industrial sectors. Such bulk enzymes are produced
primarily by bacteria by members of the general Bacillus.Fungal α- amylases most commonly
used industrially are produced by species of Aspergillus, most notably A. oryzae and only
one species of Penicillum is Penicillum fellutanum (Flaschel E, Bott M et al. 2006). A few
industrially important enzymes may be obtained from yeast, mainly from species of
Saccharomyces. Amylase growth and production by Bacillus sp. WN11 was studied under
different cultivation conditions. Maximum growth was observed at 65 °C while the highest
amylase production was recorded at 55 °C. Accounted amylase production and growth was
observed at an initial medium pH of 5 and 6. Growth was not detected, and observed at pH 4
and 9. The organism grew better and produced higher levels of amylase activity using
proteose peptone and tryptone as nitrogen sources. Growth was not observed, when NO3+,
NH4+ and urea were used as nitrogen sources. Among defined carbohydrates, tested starch
and maltose supported good growth and amylase production, with the highest productivity
recorded in the presence of starch (24 U/ml) (Mamo and Gessesse 1999).
Amylase substrate
The starch as substrate represents the most abundant storage of polysaccharides in plants, and
next to cellulose, is the most abundant polysaccharide found on earth. The starch polymer
consists exclusively of glucose units. Two forms exist, namely α-amylose and amylopectin.
α-amylose is along, linear polymer in which successive D-glucose molecules are linked by an
α 1 to 4 glycosidic bond. Individual α-amylose chains may vary in length and hence in
molecular masses in the region of 500 kDa. Abundant supplies of starch may obtain from
seeds and tubers, such as corn, wheat, rice, tapioca and potato. The widespread availability of
starch from such inexpensive sources, coupled with large-scale production of amylolytic
enzymes, facilitates production of syrups, containing glucose, fructose or maltose, which is
considerable importance in the food and confectionery industry (Elmarzugi, Enshasy et al.
2010). The initial step in starch hydrolysis entails disruption of the starch granule.
Solubilization of the granules, the process of “gelatinization” facilitates subsequent catalytic
degradation. Gelatinization is normally, achieved by heating the starch slurry to temperatures
in excess of 100°C for several minutes. α-amylase may be added immediately prior to the
heating step, in order to render more efficient the process of granules disruption. Once the
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granules have been disrupted, additional α-amylase is added in order to liquefy the starch
This process reduces the viscosity of the starch solution, and this can be shows in figure 4.
Figure 4. Controlled hydrolysis of starch, yielding up to a 96% glucose syrup.
Starch may be enzymatically hydrolysed using varying combination of amylolytic enzymes
in order to generate specific end products. The major end products produced commercially
are maltodextrins or corn syrup solids, maltose syrup, glucose syrup or high fructose corn
syrup, and cyclodextrins. Maltodextrins are produced by partial enzymatic hydrolysis of
starch, using α-amylase. The product is often defined as a non sweet nutritive saccharide mix,
largely containing glucose units, which is linked primarily by α1 to 4 glycosidic linkages.
Although the exact product composition can vary, it generally consists of glucose (approx.
1%), maltose (3-5%), maltotriose (5-10%), maltotetraose (6%), with the reminder (75% or
more) being saccharides of higher molecular mass (Rodríguez, Alameda et al. 2006). When
the desired degree of starch hydrolysis attained by α-amylase, the pH value of the slurry
dropped from 6.5 to 3.0 and the slurry, is then heated to boiling for about five minutes in
order to totally inactivate the α-amylase. Filtration and passage through a carbon column
removes any particulates and colour, and the resultant purified maltodextrin mix is
evaporated and subsequently spray dried to yield a powdered product. The reaction rates of
amylase and starch increased as the temperature increased. The absorbance levels of the
starch decreased as the temperature increased. However, after 65 degree trial, the reaction
rates starts to decrease and the absorbance levels increased (Nahas E and Waldemarin MM
2012).
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Starch digestion in humans
The action of α-amylase on starch in the human body is digesting starch and producing
shorter chain glucose polymers, which are very similar to the action of α-amylase outside
human body. In humans, the salivary and pancreatic glands produce α-amylase. However, the
action of salivary amylase is limited to the hydrolysis of starch to lower degree of
polymerization polymers. The principal hydrolysis of starch by pancreatic α-amylase occurs
within the lumen of the small intestine; this action produces shorter chain polymers of lower
average polymerization degree than the intact starch molecule. The lower degree of
polymerization polymers are subsequently act on by other enzymes, including glucoamylase,
which cleaves the polymers into individual glucose monomers residues that are absorbed by
the body (Ai, Nelson et al. 2013, Elmarzugi, Enshasy et al. 2013).
The impact of genetic engineering
Genetic engineering continues to have a major impact upon the industrial enzyme sector.
Throughout the 1980s the major enzyme companies initiated research programmed aimed at
producing selected industrial enzymes by recombinant technology. Recombinant strategies
using both homologous and heterologous expression systems. Production of industrial
enzymes by recombinant means displays a number of potential advantages, as compared to
traditional (non-recombinant) approaches. These advantages may include the higher
expression levels attainable; product generally displays a higher degree of relative purity.
Moreover, it is considered economically attractive. Heterologous expression facilities
commercialization of enzymes produced naturally by pathogenic/ non-GRAS-listed species,
also, allows alteration of enzyme’s characteristics via protein engineering. The advent of
recombinant DNA technology has facilitated the cloning and expression of genes coding for
various α-amylases in a variety of recombinant organisms. Human, wheat and bacterial α-
amylases, for example, been expressed in saccharomyces cerevisiae. More recently, the gene
coding for B. licheniformis α-amylase has been expressed in transgenic tobacco plants, this
was among the first examples of the production of bulk industrial enzymes in a recombinant
plant species. Microbial amylases could be potentially useful in the pharmaceutical and fine
chemical industries if enzymes with suitable properties could be prepared. The molecular
mass of the recombinant protein was found to be 64 kDa, compared to 55 kDa in the native
Bacillus species (Deb, Talukdar et al. 2013).
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Applications
α-Amylase in textile de-sizing
Initial moves to apply biotechnology have been predominantly in textile finishing.
Biotechnological processes have already proved more advantageous than chemical processes
in the processing of natural fibres, such as cotton and wool. Modern methods of textile
weaving, places considerable mechanical stress on the fabric threads to prevent breakage, the
fabric strands are nearly always coated with a substance known as a ‘size’. The serves to
strengthen the fabric prior to weaving. Essential attributes of sizing materials include good
adhesion to the textile threads, ease of removal after weaving, it is necessary to remove the
size after weaving as it would subsequently prevent proper dyeing or bleaching of the
finished product and inexpensiveness. Subsequent removal of the starch (i.e. ‘desizing’) may
be achieved by steam heating in presence of NaOH, or by oxidants. However, such
treatments can damage the textile and will generate a process effluent which must treat before
disposal. Desizing using α-amylase has thus become popular. Generally, thermo-stable
bacterial α-amylases (e.g. B. licheniformis α-amylase) are mainly used. Depending upon the
enzyme concentration and environmental parameters chosen, the desizing process may last
from several minutes to several hours (Walsh G., 2005; Flashel E. et al., 2006). The
European textile industry is exposed to fierce international competition and price wars. In the
last few years this has caused parts of the European textile production to migrate to Asia. As
the European textile industry cannot compete with Asian complying with respect to wages
and the cost of complying with environmental restrictions it is dependent on initiating new
products and new production processes, for example technical textiles (textiles with
customized and functional properties). High growth rates are particularly anticipated for
technical and functional textiles, e.g. textiles for persons with allergies, antimicrobial textiles,
textiles for hospital hygiene and those incorporated into cosmetics and pharmaceuticals. Like
textiles, pulp and paper are also often sized using starch. Sizing of paper protects it from
mechanical damage during manufacture, and enhances the stiffness and feel of the finished
product, in this case desizing is not subsequently carried out. Natural unprocessed starch
slurries are too viscous to be used in paper sizing. α-amylase is used to partially degrade the
starch in order to yield a product of appropriate viscosity for the task at hand. The world
market for enzymes for use in the pulp and paper industry has a volume of around € 10m and
an estimated annual growth 25 % (Longyun Hao, Rui Wang et al. 2013). As bio-
technological processes have proved more advantageous than chemical processes, 80 % of
the European textile industry now uses the biocatalyst amylase to remove starch from natural
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fibres (e.g. cotton) during the desizing process. In various areas of textile industry the
increasing application of biotechnological processes as in sourcing cotton has enabled the
branch in the short to medium term to produce more competitively and with less pollution
and to develop more innovative products (Flaschel E, Bott M et al. 2006).
α-Amylase in detergency (bio-washing)
Enzyme is used in cleaning product as cleaning and fabric care agent. Most of the used of
enzyme types breakdown large water insoluble soils and stain which attached to fabrics into
smaller, therefore removed by mechanical action of the washing machine or by interaction
with other detergent agent. Enzyme does not loose its functionality after having worked on
one stain and continues to work on the next one. The most important reason to apply enzyme
in detergency is the use of very small quantity of these bio-catalysts that can replace very
large quantity of chemical detergent (Mitidieri, Souza Martinelli et al. 2006). Amylases are
used to remove residues of starch-based foods like potatoes, spaghetti, custards, gravies and
chocolate. This type of enzyme can be used in laundry detergency as well as in dishwashing
detergency. α-amylases have been used in detergents since the beginning of the 1970s and the
demand is still increasing. The use of a such enzyme could reduce energy consumption, since
washing cycles can be carried out at 30-40°C instead of 80-90°C and less cycles are
necessary, and consequently reduced water consumption (up to 50%). Moreover, a
distinguish saving by enzymatic removal of impurities from cotton fibres, and by using
catalases for hydrogen peroxide removal after bleaching, so a decrease in the amount of
chemicals used and a decrease in the pollution due to reduced wastewater (Roy, Rai et al.
2012). The detergent α-amylases in use today derived mainly from the Bacillus family, have
highly homologous primary amino acid sequences and a tertiary structure comprised of three
domains A, B and C. The active site is typically located in a cleft between domains A and B
and is usually comprised of acidic amino acids such as aspartic and glutamic acids. Like
many proteases, most α-amylases contain essential calcium ions, which are important in
maintaining the tertiary structure of the enzyme molecule. In 2004, a new amylase,
Stainzyme (Trade Mark), was introduced which shows a significantly stronger performance
effect in most detergent segments compared to other traditional commercial amylases. This
enzyme has a broader pH and temperature range as compared to earlier amylases, meaning
that in a typical detergent, for example, nearly equal performance is obtained at both 30◦C as
at 40◦C. Furthermore, the washing time can also be reduced but still nearly, the same wash
performance will be reached (Roy and Mukherjee 2013).
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α-Amylase in food industry
α-amylases have been approved and used for many years in food manufacture. Approval of
the use of this enzyme has advantages for food manufacturers by providing a different source
of the α-amylase enzyme. The first, which has greater thermal stability and produces a
different sugar profile. There are no significant disadvantages to food manufacturers,
consumers or government agencies. In 2003 the German food industry employed around
525,000 workers in approximately 5,880 businesses and had a total turnover of around € 128
billion. However, similar to other branches of German industry, it faces increasing
international competition. Innovations could be the key to new product areas. Therefore,
experts take an optimistic view of growth in the area of functional food (Sá FC, Vasconcellos
RS et al. 2013). The world market volume for functional food could rise to about US$ 10 to
22 billion. Future prognoses anticipate annual growth rates in the world market in excess of
20 %, one reason for the increasing demand could be that many food products today, such as
chocolate pudding, ketchup, spaghetti sauce, chilli sauce, fruit puree and baby food, contain
starch in different forms (modified starches) in order to get, among other things, appropriate
viscosity (Flaschel E, Bott M et al. 2006). In food production, enzymes have a number of
advantages:
They are welcomed as alternatives to traditional chemical-based technology, and can
replace synthetic chemicals in many processes. This can allow real advances in the
environmental performance of production processes, through lower energy consumption and
biodegradability
They are more specific in their action than synthetic chemicals. Processes, which use
enzymes, therefore have fewer side reactions and waste by-products, giving higher quality
products and reducing the likelihood of pollution. They allow some processes to be carried
out which would otherwise be impossible. An example is the production of clear apple juice
concentrate, which relies on the use of the enzyme, pectinase (Marrelli M, Loizzo MR et al.
2013).
Therapeutical and Diagnostic application of α-amylase
α-amylase is widely used in pharmaceutical industry in various digestive aid preparations.
Due to presence of bacterial α-amylase, starch in the consumed food is better digested and
increase overall digestibility of food. Such digestive aid preparation is used for treatment of
patient whose digestive power is reduced due to certain illness. Many such commercial
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formulations of digestive aids are seen in drug stores either as syrup or as tablet (Faulks and
Bailey 1990). The first clinical gene therapy is underway to correct an enzyme deficiency
called Adinoseine deaminase (ADA) in children. There is a correlation between salivary α-
amylase activity and pain scale in patients with chronic pain. The visual analogue scale
(VAS) is commonly used to assess pain intensity. However, the VAS is of limited value if
patients fail to reliably report. Objective assessments are therefore clearly preferable.
Previous reports suggest that elevated salivary α-amylase may reflect increased physical
stress. There is a close association between salivary α-amylase and plasma norepinephrine
under stressful physical conditions. In this study, we have determined the usefulness of a
portable salivary α-amylase analyzer as an objective biomarker of stress (Subramani and
Aalbersberg 2012).
Enzymes are found in all tissues and fluids of the body. Intracellular enzymes catalyze the
reactions of metabolic pathways. Plasma membrane enzymes regulate catalysis within cells in
response to extra cellular signals, and enzymes of the circulatory system are responsible for
regulating the clotting of blood. Almost every significant life process is dependent on enzyme
activity. α-amylase production in aqueous two phase systems by another species of bacteria
Bacillus in polyethylene glycol (PEG 10 000)/dextran (505 000) aqueous two-phase systems
at various concentrations. An increase in the PEG concentration from 7 to 25 % (w/w) in the
aqueous two-phase system resulted in an increase in the phase volume ratio with a
concomitant decrease in the partition coefficient (K) and recovery of α-amylase in the top
phase. However, varying dextran concentrations from 2.5 to 10% (w/w) decreased both the
phase volume ratio and the partition coefficient of α-amylase at dextran concentrations lower
than 2.5% (w/w) the phases could not separate (AKCAN 2011). Bacillus species have
traditionally been utilised in the production of industrially important enzymes for a number of
reasons: with the exception of the Bacillus cereus group, they are all generally recognized as
safe (GRAS) listed and they are widely distributed in nature, they are easily cultured in
relatively inexpensive media, and members of the genera Bacillus are capable of producing
large quantities of many desirable enzymatic activities, most of which are secreted
extacellularly into the fermentation medium. Extra cellular production of enzymes obviously
simplifies subsequent product recovery and purification. Enzyme therapy is an effective safe
alternative way in cancer treatment by reversing the tumour from malignant to benign. This
could be applied, through its function. Firstly, by restore the body internal environment; this
includes blood pH to be slightly alkaline, eliminate body waste, also restore intestinal
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bacteria balance, strengthen the immune system, improve digestion and facilitate execration.
Secondly, enzyme has decomposition effect through breaking down the fibrin coating of the
cancer cell to unmask it in order for the immune system to identify as (non-self) and attack
and destroy it. Thirdly, throughout blood purification by eliminate wastes decompose toxin;
maintain right blood pH and fluidity; facilitated blood circulation when blood circulate well
so it can transport oxygen and nutrient to cell and cancer cell can not live in oxygen rich
environment. The enzyme therapy has a valuable advantage on conventional treatment; that
enzyme therapy neither treats the underlying cause and the whole body nor the symptom,
therefore it is considered a natural, non toxic, no side effects and easy to use.
CONCLUSION
Enzymes are tools of nature using natures’ own technology in modern industry associated
with many advantages, compared to chemicals. Amylases have a great commercial value in
biotechnological applications ranging from textile, bio washing, food, therapeutic and
pharmaceutical industries. These uses have placed greater stress on increasing amylase
production and search for more efficient processes.
ACKNOWLEDGEMENT
The authors would like to thank the following students Elrayes H. A., Mohamad M. S., Naji
S. A. for their assistance.
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