ENZYMES IN BAKING

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The estimate made by the Ministry of Food Processing Industries (1998), the total
market of bakery products, bread and biscuit is estimated at 1.5mn ton and 1.1mn ton
respectively. The cake market is estimated at 0.4mn ton. The organized segment of the
biscuit market is estimated to be 0.44mn tons whereas the unorganized sector accounts
for the balance 0.66mn tons. Bread market is estimated to be growing at around 7% pa in
volume terms, whereas the biscuit market in the recent years has witnessed a little higher
growth at around 8-10% pa. Within the biscuit category, cream and specialty biscuits are
growing at faster pace at 20% pa, while the popular segment is growing.

Industrial Enzymes

Industrial enzymes business has been growing in India. It was estimated to have touched Rs 375
crore with growth rate of 17 percent for the year 2005-06. Novozymes and Biocon contribute over
50 percent of the total enzymes revenue. Biocon is leading the Indian enzymes sector with Rs 85
crore (2005-06) followed by Novozymes. The other leading companies in this sector include
Advanced Enzymes Technologies, Rossari Biotech, Maps India, Zytex India, Lumis Biotech and
Textan Chemicals.

There are about 15 companies involved in enzymes business with few into manufacturing. The
rest are into the marketing business. Although the sector has been on growth trajectory, with no
regulatory issues it is still to get the attention of the entrepreneurs. The sector saw no new
companies during the year. It is because of high initial cost involved in setting up the facility and
also lack of skilled people with technical know how. Only handful of companies like Biocon,
Advanced Enzymes, Lumis Biotech, Maps India, Textan Chemicals, Rossari Biotech have
established manufacturing facilities in India. The rest are involved in the formulation and trading
activities. Indian companies are manufacturing microbial enzymes but still look at importing
glandular enzymes. The companies have been importing enzymes from USA, Canada and China.

The companies are now looking for newer areas like food processing, agriculture, animal
nutrition, dairy, aquaculture and marine besides the conventional areas such as pharmaceuticals,
textiles, breweries, chemicals, and tannery etc.

Despite the sector witnessing a growth of 17 percent, Biocon's sales from enzymes have
registered a decline of 5.3 percent during the year due to capacity constraints. The increased
capacity consequent to the setting up of the new facility at Biocon Park is expected to contribute
to the growth of enzymes in the coming years. Biocon develops and markets a mix of specialty
and industrial enzymes for a broad range of industries. Biocon, the first enzyme company globally
to receive the ISO 9001 accreditation, and India's largest producer, supplies enzymes to over 50
countries across a wide range of industry segments including healthcare, beverage, brewing,
baking, fruit juice, starch & alcohol, sugar, animal health, paper & pulp, leather and detergent.

Novozymes, a multinational involved in marketing of enzymes through its distributors in India, has
successfully introduced the enzymatic degumming of rice bran oil. The leading rice bran oil
refining companies in India have been using the enzymatic degumming process that minimizes
losses of oil, produces consistent degumming results, reduces water consumption, and is more
environmentally sound. Novozymes's baking enzymes Fungamyl, Pentopan, Lipopan, Gluzyme
and Novamyl used to improve western-style bread have received good response from the
customers. Similarly Lumis Biotech, a flagship company of Rs 160 crore Kgenix group has
launched its indigenously developed peroxide killer used for removing residual peroxide from the
bleach.

Advanced Enzyme Technologies (formerly Advanced Biochemicals), another leading enzyme

manufacturer, has consolidated its position during the year by increasing its sales revenue
registering a growth rate of 41 percent. It is expected to increase its growth rate during the year
since two of its new manufacturing facilities, one at Indore in Madhya Pradesh and other at Thane
in Maharashtra, would be operational during the current financial year. Advanced has invested Rs
45 crore in these two facilities besides setting up an R&D facility.

For Advanced Enzyme, last year was a year of development. It focused on product and process
developments. It successfully complied for the SKAL International Audit for Organic Certification.
With this, it joined the SKAL International Program for Organic Certification for its agriculture
product AgroSEB, an organic bio enzyme.

Rossari Biotech, an ISO 9001: 2000 company and manufacturer of enzymes for textile
processing, has started new concept of enzymatic bio-scouring. To meet the growing demand, it
imports enzymes from the US and supplies the formulations to the Indian companies. It has
increased its number of customers too during 2005-06.

During 2005, Lumis Biotech received approval from GEAC to import Denimax 399S, Denimax
16000 L, Denimax Acid XCL and Aquazyme Ultra 1200 L from Novozymes, Denmark with couple
of conditions for value addition and export purpose.

Considering the potential and opportunity in enzymes, few companies are expanding their
manufacturing facilities so that they can also export their range of products. The government of
India has also been considerate to the sector, as it has been funding many projects related to
industrial enzymes. This will help the industry to growth and compete with the global players by
launching new products in near future.

BAKERY LAGS BEHIND DAIRY. Bakery products are consumed virtually everywhere across the
globe and make up more than 20% of all packaged food sales. Worldwide, the sector was worth
€245 billion (US$311 billion) in value sales in 2005, exceeding those of the dairy sector at €223
billion (US$283 billion). And yet, where fortified and functional products are concerned, bakery is
lagging behind. Fortified/functional bakery products accounted for less than 2% of total bakery
value sales, compared with dairy’s 7.5%. Dairy has also beaten bakery in the growth stakes —
31% for fortified/functional dairy compared with 20% for fortified/functional bakery during 2002-05.
Unlike the dairy sector where fortified/functional products are found virtually throughout, most of
the bakery sector’s are concentrated in one subsector — breakfast cereals — which accounts for
65% of sales.

Dairy manufacturers were remarkably quick at capitalizing on their products’ naturally healthy
image as an excellent source of calcium and vitamins. In many countries, including the US, dairy
producers started to routinely fortify milk back in the 1930s, in line with a governmental public
health policy geared at eradicating malnutrition. The dairy industry has built on this, by further
fortification, innovation and promotion.

In contrast, the bakery industry was slower to catch on. However, the bakery sector is in no way
inherently disadvantaged here — many bakery products are also considered a naturally healthy
staple food, and people have been benefiting from their fortification for decades. In the UK, the
fortification of white flour with thiamine, iron, niacin and calcium is mandatory. In the US
enrichment of white flour and bread has been mandatory in many states in the 1950s and
nationwide since 1993. Folic acid was added to the mandatory mix in 1998 to help prevent neural
tube defects in newborns.

However, it took the arrival of the Atkins Diet to truly jolt the bakery world out of its slumber where
fortification is concerned. Suddenly, bakery foods were "evil" and full of fattening carbohydrates.
Frantic innovation was the result of the fad, and sales of reduced-carb bakery items grew five-fold
between 2002 and 2005. The fad quickly died out, but left the industry with the realization that
bakery could no longer rely on either its staple or indulgence status. The global health-and-
wellness trend was gathering momentum, consumers needed to be courted, and market share
fought for. The only way forward for bakery was innovation.

PUTTING BACK THE ROUGHAGE. Fiber is a natural component of grain and conveys many
health benefits, from regulating bowel movements and increased levels of satiety to reducing
cholesterol levels and possibly even preventing bowel cancer. Much like the dairy industry turning
away from sterilizing its yoghurts and going "bio live," the baking industry is putting the roughage
back in. Indeed, fiber is one of the main global growth drivers of health-and-wellness baked
foods. In the US, high-fiber breads grew by 15% in 2005, and Euromonitor International predicts a
further increase by 40% between 2005 and 2010.

Soluble fiber has been elevated to advanced functional heights, especially since one of its
components, beta-glucan, was approved for heart health claims by the US Food and Drug
Administration in 1998, reviving the 1980s oat craze. The next big thing on this front will be
barley, which also contains beta-glucan. In May 2006, the FDA extended the heart health claim to
barley. Cargill, Minneapolis, MN, USA, recently introduced Barliv barley beta-glucan, an
ingredient marketed as cholesterol-reducing.

PLANT STEROLS AND OMEGA-3 OILS. The most remarkable innovation of recent years was
the launch of cholesterol-reducing functional spreads with added phytosterols, introduced by
Raisio and Unilever. Although these products are not doing well in the US, they are a hit in
Europe, and the dairy industry, was the first sector to benefit — successfully integrating
phytosterols and stanols into drinking yoghurts and other products.

Bakery is now following suit. Earlier this year, Finnish-owned Fazer Bakeries was given the go-
ahead by the European Commission to market its sterol-enriched rye bread. In the US, Cargill
announced in June 2006 that Bimbo’s Oroweat brand was the first to contain Cargill’s CoroWise
plant sterols. Such products will do well on the European market and, with sufficient promotion,
should also appeal to the ageing US population.

An even bigger phenomenon is the omega-3 craze, which also originated in the oils and fats
sector. Although omega-3 fatty acids are not naturally associated with dairy products, the dairy
industry translated this trend into product launches, and omega-3-enriched milks are now on the
market.

Bakery can count itself lucky where omega-3 is concerned: Flax seed is absolutely packed with it
and relatively easy to integrate into breads. European bakers have a definite advantage here —
German, Austrian, Swiss and Scandinavian consumers, among others, traditionally appreciate
heavy breads spiked with an assortment of seeds. FDA duly approved a health claim pertaining
to omega-3, and bakery manufacturers and retailers, including Wegmans Food Market, Bimbo
Bakeries USA, The Baker and Arnold Foods Co., have launched omega-3 fortified breads in the
US.

Advances in omega-3 production from fish oil have greatly helped this trend, and things will be
heating up even more: Martek Bioscience Corp., Columbia, MD, USA, announced in June 2006
that it struck a 15-year deal with General Mills, Minneapolis, MN, USA, with the aim of integrating
Martek’s omega-3 oils into as many bakery products as possible. Another June 2006
announcement of the joint marketing and development partnership between Puleva Biotech,
Granada, Spain, and The Wright Group, Crowley, LA, USA, shows the industry’s commitment to
creating new products and markets for omega-3s.

FERTILE FUTURE IN THE EAST. The US is undoubtedly the most advanced market in terms of
fortified/functional breads and biscuits, accounting for 34% and 61% of global sales, respectively.
However, the market is still in its infancy and the potential and innovative drive of other countries
is not to be underestimated. Spanish manufacturers, for example, are very active on the
fortified/functional bakery front. Adult-oriented biscuits are starting to flourish there, starring a line-
up of sophisticated ingredients such as digestive enzymes, antioxidant-rich green tea and, of
course, plant sterols and fiber.

In Eastern Europe, equally interesting developments are starting to take root. The Hungarian
fortified/functional bread market is worth €11.8 million (US$15 million), topped by Gyori Süto’s
Seleno, a selenium-enriched bread. Eastern Europe is a developing market, and
fortified/functional bakery is a tiny segment, but its potential is significant. The global health-and-
wellness trend has given rise to better-for-you

Bakery and Confectionery

Introduction
The estimate made by the Ministry of Food Processing Industries (1998), the total market of bakery product,
bread and biscuit is estimated at 1.5mn ton and 1.1mn ton respectively. The cake market is estimated at 0.4mn
ton. The organized segment of the biscuit market is estimated to be 0.44mn tons whereas the unorganized sector
accounts for the balance 0.66mn tons. Bread market is estimated to be growing at around 7% pa in volume
terms, whereas the biscuit market in the recent years has witnessed a little higher growth at around 8-10% pa.
Within the biscuit category, cream and specialty biscuits are growing at faster pace at 20% pa, while the popular
segment is growing.

Production area
Besides the industrial areas in leading metropolis the bakery product & confectionery are carried on small-
scale basis also at household level. Whereas, the confectionery industry has developed remarkably with the
international brand mingling with the domestic market toffees, chocolates etc. produced at large scale in
important industrial regions of the country.

Growth promotional activities

In 1977- 78 Government reserved the confectionery bread and biscuit manufacturing for small scale and
restricted entry of large producers. During the last 2 decades, small and unorganized players shared the growth
in the industry. Currently, there are an estimated 2 million bakeries across the country engaged in production of
bread, biscuits and other products The Indian confectionery market is segmented into sugar-boiled confectionery,
chocolates, mints and chewing gums. Sugar-boiled confectionery, consisting of hard boiled candy, toffees and
other sugar-based candies, is the largest of the segments and valued at around Rs 2,000 crore.

The confectionery industry has a current capacity of 85, 000 tonnes, the market is growing at the rate of 10-
15% per annum. The estimated annual production of bakery products in India is in excess of 3 million tonnes, of
which bread accounts for nearly 50% and biscuits 37% in volume terms in the organized sector. The bakery
sector in India is one of largest segments of the food processing industries; annual turnover in value terms is
approximately $ 900 million.

Types
Chocolates, hard-boiled sweets, chewing gum and other products are a major growth area. The bakery industry
includes basic baked products like breads, biscuits, cakes, pastries, rusks, buns, rolls etc.

Though the usage of enzymes in an industrial scale in the food industry is a recent phenomenon,
the usage of enzymes in food preparation per se is hundreds of years old. For generations, enzymes
have been used for the improvement in texture and appearance, enhancement of nutritional value
and generation of appealing flavors and aromas. Currently the applications for enzymes can be
found in bakery, cheese making, starch processing and production of fruit juices and other drinks.

Among the enzymes used in food applications, those used in bakery industry constitute nearly one
third of the market. The market growth rate is the highest for the bakery enzymes among all the
food enzymes. The ever expanding applications of the enzymes in the bakery segment are the
reason behind the fast growth. The bakery products have undergone radical improvements in
quality over the past ten years in terms of flavors, texture and shelf-life. The usage of enzymes is
the biggest contributor for these improvements.

The baking industry makes use of four types of enzymes. Amylases are used to convert starch to
sugar and to produce dextrins. For strengthening and bleaching of the dough, Oxidases are used.
Hemicellulases and Proteases are the enzymes which act on the wheat protein, gluten. While
hemicellulases improve gluten strength, proteases reduce gluten elasticity. All these enzymes
together play an important role in maintaining bread volume, crumb softness, crust crispiness and
browning and maintaining Freshness.

Among all the food applications, the usage of enzymes in the baking industry is the most
complicated. And some consider Enzyme blending in baked goods to be an art form. The blending
has to be precise in order for the enzymes to effectively and consistently deliver the desired
benefits. This requires some planning and a lot of understanding on the part of the designer of the
blend. The blender should be well versed with the effects of factors like temperature, acidity of the
dough and time given for the chemical reaction.

Genetic engineering: opportunities missed

In the early 1980s, the enzyme industry started to exploit the developments taking place in the
fields of biotechnology and genetic engineering. This meant a rapid and radical development in the
enzyme production technologies. The potential to improve the production efficiency and the product
quality of enzymes through genetically modified organisms is immense. But the adoption of GMOs
by the baking industry can hardly be described as smooth. General perception among the public for
GMOs is negative.

This attitude of consumers towards GM products has been a major factor for companies to

go slow on full conversion to more economically productive biotechnology-based manufacturing

processes. In this aspect the consumer perception in the US is highly influenced the consumers in
Europe, who are deeply suspicious of GMO’s/GM foods. The enzyme manufacturers in the US have
also contributed with their inability to convince the public of the safety of genetic modification. The
GM controversy is holding up enzymes with very promising potential in baking applications.

Chemical processes set to go the Bromate way

Enzymes are fast replacing many chemical processes in the baking industry. The rapid decline in the
usage of bromates is a case in point. Potassium Bromate was the most widely used volume
enhancer and dough strengthener. Since bromate’s carcinogenic effects were made public in 1982,
its usage world over has come down drastically. Countries like the UK and Canada have completely
banned its usage. Though there is no legal ban on the use of bromate in the US, due to the
increasing health awareness, both the consumers and the baking industry have been averse to its
usage. The enzyme industry has provided an effective replacement for bromate. The enzyme alpha-
amylase in combination with Ascorbic acid is the most used substance to improve the volume and
the quality of the crumb. There are many custom blends available in the market with optimized
mixture of ascorbic acid and Alpha-amylase.

Enhanced shelf-life breathes life into the industry.

The single biggest contribution of enzymes to the baking industry is the enhancement of shelf-life.
Most of the bakery products undergo a progressive deterioration of quality when stored at ambient
temperatures beyond a point of time. This deterioration is termed as staling. Enzymes slow down
the staling process by improving the moisture retention in baked products. The addition of dough
strengthening enzymes like Alpha-amylase will create optimally developed and functional protein
structure. This results in a softer crumb and considerable increase in the shelf life of the final
product. Industry experts feel that the improvement in shelf life is one of the biggest growth drivers
for the bakery enzyme industry.

Markets : Bigger players hold the sway.

The major companies in the bakery enzyme market are of three types.

Manufactures such as DSM produce enzyme for both captive use and free market sales. Some
manufacturers focus on selling to end-users and resellers. Then there is a category described as co-
producers or blenders, who supply specialized blends to the end-user industry based on specific
requirements.

The Current market leaders in the US markets for Bakery enzymes are Novozyme, Danisco and DSM
in that order.

This is a business based on product innovation and the ability to innovate is proportional to the
investment a company can make in R&D. Since R&D is expensive to cultivate and also maintain,
only large companies with significant resources and strength in the different functional areas will be
able to remain competitive.

Enzymes:
Valuable Assets
September 1998 -- New Technologies

By: Suanne J. Klahorst

Contributing Editor

Enzymes - the biological catalysts regulating the chemical reactions necessary to sustain life - are
regarded either as problems or solutions, depending on their impact on food processing and product quality.

For the fresh-cut fruit and vegetable processors, endogenous enzymes from plant tissues
are responsible for browning, adverse flavor changes and texture loss - changes that need
to be avoided by blanching, chilling, freezing, acidification or sterilization. In a bakery
operation, enzymes are viewed as valuable assets that make the job of turning out
consistent bakery products a little easier. Historically, malt extracts - which are rich in
native barley enzymes - were added to doughs to get the benefit of those enzyme
activities. Today, it is common to supplement native flour enzymes with exogenous
enzymes produced by microorganisms, particularly amylases, proteases and xylanases.
Some of the benefits of enzymes in bakery products are consistently improved
machinability, higher loaf volume, smoother crumb characteristics and longer shelf life.

Almost all food products are made from living systems, so all foods are subject to at
least one, but typically dozens, of enzymatic reactions. That's why enzymes represent not
only the most challenging problems, but also provide the most elegant solutions in food
product and process design.

Promising proteases
Enzymes are commonly named for the substrates they catalyze, with a few exceptions. Their
purpose is to enable a reaction that alters the chemical structure of that substrate, indicated by
the added suffix "-ase." Enzymes that hydrolyze protein are termed proteinase, or more
commonly, protease. While proteases are specific for protein, they are quite versatile in their
capacity to hydrolyze a variety of protein bonds from different sources - both animal and
vegetable. For each type of enzyme and each type of protein, the end-products of the reaction
are different. In the case of the specialized mammalian protease called chymosin, a limited
hydrolysis of milk protein is achievable for the specific purpose of forming a protein curd for
cheesemaking. Other types of proteases are able to convert solid proteins from meat, fish or
legumes into liquid slurries or protein hydrolysates, as in the production of savory flavors. Some
proteases are effective at hydrolyzing proteins all the way to amino acids.

Although a wide variety of bacterial and fungal proteases are now available, papain still
enjoys a certain popularity as a natural plant enzyme that can achieve results similar to
those of the high-tech microbial proteases that threatened to make it obsolete. Papain has
enjoyed a market in several industries, including brewing, where it has been used for
chill-proofing beer via its ability to degrade the protein component of haze.

Papain has its limitations, however. Most papain is imported from regions of the world
where a special variety of papaya fruit is grown for its protease enzyme content. Papain
importers usually have very little control over the enzyme extraction and purification
process. Quality can vary as a result.

Valley Research, Inc., an enzyme company based in South Bend, IN, finally eliminated
two limitations of papain. "Papain has an odor reminiscent of old gym socks," explains
President Art Sears. "After two years of working with papain growers in India, we
developed an improved enzyme-recovery process that not only results in an enzyme
product with less odor, but eliminates the use of metabisulfite as a preservative." Valley
Research also is marketing another plant protease for food processing. Called "actinidin,"
it is extracted from kiwi fruit.

As the selection of proteases from various plant, animal and microbial sources increases,
the opportunities for combining enzymes to create new food ingredients multiply.
Proteases, in particular, are becoming valuable tools in the conversion of protein
byproducts into high-value functional ingredients.

Jamel S. Hamada, Ph.D., research chemist, Southern Regional Research Center,

USDA's Agricultural Research Service, New Orleans, specializes in enzymatic
modification of proteins for expanded food uses. He currently collaborates with
Louisiana State University in the identification and development of new uses for rice
bran. "Rice bran is sold for as little as 2 cents a pound, but the bran contains from 12% to
20% protein," Hamada says. "If these proteins could be recovered and solubilized, the
protein value could be considerably higher." By combining two food-grade proteases
from Novo Nordisk, Basvaerd, Denmark, Hamada demonstrated that he could recover
proteins from rice bran and improve them to the extent that they exhibited emulsifying
functionality similar to casein. He presented the results of his ongoing research at the IFT
Annual Meeting in June.

Two are better

Enzyme companies specialize in leveraging the synergistic effects of more than one enzyme to
improve the functionality in food systems. One of the most complex and precise enzyme-blending
challenges is the baking industry. With specific enzymes for degrading each of the components of
flour, including starch, protein, and xylans (pentosans), the possibilities for combining all the
activities into one product are nearly infinite. In baking, enzyme blending is as much an art as a
science. Since flour composition is not consistent, enzyme blends must be fool-proof enough to
consistently deliver the desired benefits, without overtreating the dough to the point that product
quality or machinability is lost. The enzymes used in baking enzyme blends also must be
temperature-labile enough to inactivate at baking temperatures, to avoid residual changes in the
final product on the shelf.

Some enzyme combinations are targeted for a particular flour (wheat, rye, oat) or a
particular finished product, such as bread or crackers. Rye and oat flours contain sticky
xylan and glucan polymers that make processing troublesome without the use of
xylanases or glucanases. Amylases - named for amylopectin - convert gelatinized starch
to fermentable sugar, thereby improving the leavening and anti-staling properties of bread
and rolls. Fungal or bacterial protease addition results in a slacker dough and a reduction
in conditioning time for cracker production.

In spite of the art required, enzyme addition to baked goods has been simplified by the
availability of pretested blends that are developed in the research bakeries of enzyme
companies or enzyme-ingredient suppliers. The formula for these blends is always
proprietary to protect the art, but for those product developers who don't have the
resources or the expertise to run the extensive product trials, this is an acceptable
solution. Novo Nordisk has these baking capabilities in their Franklinton, NC, facility,
where they use test enzymes in baking to create a wide variety of baking products.

Greg LeFebvre, industry manager, Novo Nordisk, Franklinton, recently disclosed the
fall 1998 introduction of their newest enzyme blend in the Fungamyl® product line,
Super BR, an enzyme system designed to replace chemical dough conditioners, such as
potassium bromate.

"During the product test market, customers reported that Fungamyl Super BR
demonstrated improved bendability and wrapability in wheat tortillas, and improved loaf
volume and crust in bread," LeFebvre says. "Additionally, greater porosity in English
muffins has been observed in plant trials. These are all benefits that can be realized from
a gluten strengthener in baking."

Bromate was a cheap and effective dough strengthener until its good reputation was lost
after its degradation products were found to be carcinogenic to rats. Europe and Canada
responded by banning bromates, and although legal within specified limits in the United
States, California law requires a warning label if bromate is used as an additive.

Since enzymes that are inactivated in the baking process don't appear on the ingredient
label, bakeries that use bromates or chemical bromate replacers, such as
azodicarbonamide and iodates, can clean up their label by replacing some of the chemical
additives with enzyme blends. "During the product test market, customers reported that
Fungamyl Super BR created more porosity in English muffins, improved crust
appearance in hearth breads, and improved bendability and wrapability in wheat
tortillas," LeFebvre reports. Fungamyl is produced from the fungal organisms Aspergillus
niger and Aspergillus oryzae, which have been used for several years to produce enzymes
for the baking industry.

Enzyme expertise
There are many success stories for new enzyme applications and product uses. The most
encouraging fact is that many of the ideas for enzyme blends and new ways to use them originate
with the enzyme user rather than the supplier. Food scientists and technicians who gather an
arsenal of enzyme samples can create unique products and proprietary processes that enzyme
vendors cannot, because enzyme vendors lack access to seasonal raw materials and familiarity
with the customer's process.

New enzyme applications require multiple trials and determination as much as technical
enzyme expertise. Mike Turner, fruit and wine industry manager, Valley Research,
provides an illustration of how enzymes can be blended for maximum benefit. In the
juice-processing industry of the Pacific Northwest, Turner became familiar with enzymes
that are used routinely for depectinizing and clarifying fruit-juice concentrates. While
employed at a firm specializing in Concord grape concentrates, Turner became frustrated
with the costly and tedious process that started at the Concord grape harvest. "Single-
strength Concord grape juice is traditionally held in cold rooms for several months to
allow the tartrate crystals to form and settle to the bottom before concentration," Turner
explains. "So, it can be months before there is any product to sell."

Turner gathered technical information from his enzyme suppliers and Oregon State
University, Corvallis, to understand how to develop a shorter, more economical process
for concentrating and stabilizing Concord grape juice. The new process he developed
hinged on a custom blend of four complementary enzymes he added during the crushing
process. The enzyme blend, a proprietary mixture of pectinase, cellulase and
hemicellulase enzymes, depectinized the juice earlier in the process and reduced the
viscosity enough to improve the removal of insoluble solids during the centrifugation,
called "decanting." The viscosity reduction and a lower percentage of insoluble solids
was enough to make concentration of the juice possible shortly after crushing.

"The tartrates formed more readily in the concentrated juice, which decreased the hold
time required," Turner explains. "After tartrate removal, the juice was concentrated
further to 70°Brix for shipping. When the concentrate from the new process was
compared with the traditional process, it had more color and higher juice yields per ton of
grapes processed.

"Reducing the hold time also eliminated the chances of contamination by yeasts and
bacteria that convert juice to wine or vinegar during storage," he says. "We had grape
concentrate ready to sell in a month rather than several months." Turner's enzyme talent
eventually landed him his current position at Valley Research. He currently shares his
processing experience with other Concord grape juice processors. His enzyme blend,
which is still proprietary, is now commercially available from Valley Research as
Crystalzyme® Concord.
 Future enzymes
Genetic engineering has created many new opportunities for producing enzymes in food crops.
Companies specializing in transgenic plant technologies are ready to exploit the vast new
opportunities for enzyme production in plants. One such company was founded in 1993 by Ray
Rodriguez, professor, department of molecular and cellular biology, the University of California-
Davis. Rodriguez, chairman of the Board of Applied Phytologics Inc. (API), Sacramento, CA,
specializes in expressing enzymes in transgenic cereal grains. "API's current focus is on rice," he
explains, "which has the smallest genome of the major cereal crops and, consequently, the best-
known genome of the cereal grains." The gene transformation in rice was completed 10 years
ago, in 1988.

Transgenic barley, another research focus for API, was not developed until 1994, but it
offers advantages because of the potential that malting operations offer for adding value
by enhanced enzyme activity. Barley is routinely malted for brewing operations, a
process that allows the endogenous enzymes to convert the starch to sugar for
fermentation to ethanol. One proposed target is the addition of a gene coding for a heat-
stable beta-glucanase in barley that could break down most of the beta-glucan during the
malting process. Beta-glucans often interfere with the separation of wort and mash during
the brewing process (called lautering). Eliminating beta-glucans upfront, in the malting
process, would reduce the need to add enzymes later during brewing. Incidentally, API
formed a business alliance with Coors Brewing Company, Golden, CO, to examine how
enzymes destabilize plant allergens, but they've yet to collaborate on brewing
applications.

Rodriguez points out that grains offer many advantages for the delivery of enzymes to
food products because they offer low-cost production systems. Harvesting, transportation
and storage are more economical with grains than with fruit and vegetable crops. For
many of the cereal grains, the secrets of transformation need to be elucidated to achieve
"genotype-independent transformation." Once the targeted gene has been successfully
inserted into a research grain, it needs to be transferred to a commercial variety for
production. When this transformation becomes routine, the development time to get the
products to the market will be reduced. Since all the cereal grains share similarities in
their genomes, API is positioned to apply their transgenic rice know-how to other
commodity grains, such as wheat and corn, in the future. They are forming alliances with
companies interested in improving grains for delivering nutritional benefits in patented
food and nutraceutical applications.

What is organic?
Although food enzymes are usually considered processing aids and, as such, are the least
visible addition to food products, there are still unresolved issues for food companies that want to
use genetically engineered enzymes and maintain organic standards. USDA's recent attempts to
propose national standards for organic products resulted in the confirmation that food produced
by modern biotechnology does not "fit current organic practices nor meet current consumer
expectations about organic," according to Agriculture Secretary Dan Glickman. That would
eliminate the genetically modified corn and soybeans from the organic menu, but where do
recombinant enzymes fit in this scenario?
 Several organic-food processors have requested verification from their suppliers that the
enzymes used in their process are not "recombinant." This statement can refer to the
microorganism that hasn't been modified with recombinant DNA techniques and the
enzymes produced for the new DNA sequence.

A few enzyme suppliers have indicated in their literature that certain products are "not
genetically modified." However, most enzyme manufacturers do not provide this
information in their product literature. If pressed, they usually disclose the required
information to customers in the interest of maintaining the customer relationship. The
role of enzymes in organic products has been quietly debated, since the microorganisms
that produce enzymes are removed before the enzymes are concentrated or granulated
into final enzyme preparations, and since the enzymes are inactivated in the final food
product. In addition, the quantities of enzymes used as processing aids are often very low,
usually in the ppm range.

No matter which sector of the food industry is selected, there is always a need to be
more aware of food enzymes and how the problems they cause in one sector might result
in a solution for another sector.

There is a significance trend towards the change in the eating habits in India and
growing demand for bakery products. As the demand for the bakery products are
increasing day by day, so also the need for the improvement in the quality of products. To
meet the product demand, as well as for health conscious people,

Enzymes play an important role in baking industry. For decades, enzymes such as malt
and fungal alpha-amylases have been used in bread-making. Rapid advances in
biotechnology have made a number of exciting new enzymes available for the baking
industry. The importance of enzymes is likely to increase as consumers demand more
natural products free of chemical additives. For example, enzymes can be used to replace
potassium bromate, a chemical additive that has been banned in a number of countries.
The dough for white bread, rolls, buns and similar products consists of flour, water, yeast,
salt and possibly other ingredients such as sugar and fat. Flour consists of gluten, starch,
non-starch polysaccharides, lipids and trace amounts of minerals. As soon as the dough is
made, the yeast starts to work on the fermentable sugars, transforming them into alcohol
and carbon dioxide, which makes the dough rise. The main component of wheat flour is
starch. Amylases can degrade starch and produce small dextrins for the yeast to act upon.
There is also a special type of amylase that modifies starch during baking to give a
significant anti-staling effect. Gluten is a combination of proteins that forms a large
network during dough formation. This network holds the gas in during dough proofing
and baking. The strength of this gluten network is therefore extremely important for the
quality of all bread raised using yeast.
ENZYMATIC BREAD IMPROVERS SEBAKE SW SERIES A COMPLETE
REPLACER FOR POTASSIUM BROMATE (CARCINOGEN)

For a long time, amylase was thought to be the only enzymes that could be used in the
milling industry. The view has changed drastically since the introduction of bread
improver has come in the market. The bread improvers are nothing but the specially
designed premix for preparing bread. Previously cancer causing chemicals such as
potassium bromate was used as a strengthener for bread. As the technology is growing
day by day, Advanced Enzyme Technologies Limited has come up with the complete
bromate replacing bread improver known as SEBake SW series of bread improvers since
Poatssium Bromate is considered to be one of the most important carcinogenic as well as
it is banned in many of the European countries.SEBake SW when used in recipe gives the
bread a golden crust colour, finer crumb, finer texture, softness, whiter crumb, higher
volume and increased shelf life. SEBake SW series of bread improver specially designed
pentosanases helps which not only improves the gas retention, but also hinder the
formation of gluten network that retains the carbondioxide gas in the dough. The
hemicellulases which facilitates the water uptake reduces stiffness and results in softer
ENZYMATIC BISCUIT IMPROVERS AS COMPLETE REPLACER FOR
SODIUM META BI SULPHITE REPLACER(CARCINOGEN)

Enzymes are being used in biscuits, crackers and cookies. Mainly hydrolytic enzymes
such as protease, specially designed pentosanases type of enzymes are used in biscuit.
In Glucose type of biscuits, fermented variety of biscuits usually sodium Meta bi sulphite
is used as strong reducing agent for splitting the inert and intra disulphide bonds of
gluten, causes the dough to fall. Since it destroys the vitamin B1 and found to be long
term effective carcinogen, We Advanced Enzyme Technologies Limited has developed
biscuit improver which gives complete replacement for sodium Meta bi sulphite in
glucose type of biscuit. It not only causes the health benefit, but also it reduces the baking
time thus saves the energy.Since the protease takes away the internal tension, it makes the
biscuit to be flat at the bottom and helps in uniform colour development .It also helps in
giving better shine and finished texture, proper browning to the baked biscuits In case of
fermented type of biscuit; biscuit improver not only improves the characteristics of
biscuit but also reduces the fermentation time.

CAKE IMPROVER

SEBake cake improver is ready to use improver that can be whisked together with other
ingredients, thereby reducing the mixing time and also it increases the batter resistance to
mechanical handling before baking.It is designed for producing superior quality delicious
cakes with increased volume, spongy and soft cakes with whiter crumb.
Bread

The quality of a loaf of bread depends upon the quality of flour, the preparation and
processing of dough from the flour and the baking process. The way in which each of
these influence loaf quality will be considered.

1.Composition of flour

Wheat flour comprises approximately 68% starch, 16% water, 12% protein, 3%
pentosans and 0.5% fat.

a) Starch
The structure of starch was described in chapter 1. During milling some of the starch
granules are damaged. It is these that are more readily attacked by the hydrolytic
amylases during dough preparation. Approximately 1-2% of the starch must be
hydrolysed to provide sugar for yeast fermentation, so that the bread can “rise”.

b) Protein
These comprise mainly gluten components (about 80% of the total protein) and water-
soluble proteins (approximately 20%). The latter group of proteins include a number of
enzymes.
When mixed with water the substance referred to as gluten comprises two protein
fractions: glutenins which are high molecular weight proteins, 150-1000 Kdaltons and
gliadins which are smaller molecular weight proteins, 25-100 Kdaltons. Both protein
fractions are rich in the amino acids glutamine and proline but relatively poor in the more
basic amino acids. The better wheat cultivars for bread flour are those which have high
amounts of the high molecular weight glutenins. Both intra and inter polypeptide chain
disulphide bridges in glutenin appear to be important for the viscoelastic properties of
bread dough. The loaf volume is inversely related to the proportion of acid soluble
glutenin and directly related to the insoluble glutenin content of the flour.

The gluten proteins which are soluble in alcohol have been termed prolamins and these
are further subdivided into sulphur- rich, sulphur-poor and high molecular weight
prolamins. The structure of some of these proteins has been established (Godon,
1994).

The gluten network in the dough provides a framework to retain the air introduced during
kneading of the dough and the gas produced by the yeast fermentation, allowing the
dough to swell. Denaturation of the protein during baking contributes to the structure of
the final baked product.

c) Pentosans
These are polymers of five carbon sugars, mainly xylose and arabinose. These pentosans
produce viscous solutions in water. A major pentosan found in flour, it comprises a
straight chain of anhydro D-xylopyranosyl residues linked by a α 1-4 glycosidic bond,
with side chains of anhydro L-arabofuranosyl residues linked either to the 2 or 3 position
of a xylose residue. Possible covalent links between protein and pentosans may
contribute to the properties of bread.

d) Fats (lipids)
These contribute to the structure of the bread: defatted flour produces bread with a poor
volume.
Purified wheat starch does not contain free fatty acids but does contain lysophospholipid
(Morrison, 1995), whereas maize contains both the latter lipid and free fatty acid. The
hydrophobic part of the lipid molecules appear to lie inside helical stretches of amylose.
The amount of lipid-complexed amylose can affect the gelatinisation temperature of the
starch.

2. Preparation and processing of bread dough.

Flour, water, yeast and certain additives are kneaded to a dough. The dough contains 60-
70% water which is initially bound to protein. Within the dough there is a network of
gluten filled with starch granules, pentosans and lipids. Enzyme molecules are
interspersed amongst this network. Air bubbles are introduced during mixing and
kneading and CO2 bubbles form through the fermentation of sugar by yeast. Yeast
metabolism also produces precursors for flavour development. The air and CO2 expand
the dough and the dough will visibly grow. The rising or proving of the dough is
enhanced by a warm temperature, allowing optimum activity of endogenous enzymes
within the flour and of the enzymes of yeast.

A wide variety of baking processes exist, most of which have been automated. Three of
the dough processes used in bread manufacture are outlined in table 1 below.

Table 1. Dough: methods of processing bread dough.

Name of process Main features of process

Straight dough process Dough is fermented 1-3 hours before baking

Sponge and dough process
A “sponge” is formed from 60% of the flour
mixed with water, yeast and enzymes.
Fermentation for 3-5 hours at 30ºC. Remaining
40% flour, salt sugar and shortening added and
mixed. Fermented 20 min, divided and proved
for a further 55 min at 42ºC before baking

“No-time” dough. Dough is immediately divided after mixing and

proved for 55min at 42ºC before baking.
Requires fungal amylase, proteinase, and the
oxidising agent potassium bromate plus
ascorbic acid.

3.Baking

During baking, in the very early stages enzyme activity increases, permitting further
rising of the bread. At 50-60ºC yeast is killed and above this temperature the flour and
added enzymes are inactivated. The proteins are denatured and forms a rigid structure,
water is released from the proteins and absorbed by the starch granules. Gelatinization of
the starch occurs. Browning of the crust is due to the caramelization of sugars and
interaction between sugars and amino acids and peptides. These small molecular weight
substances are produced by the action of hydrolytic enzymes during dough proving and
early baking. In addition to the appearance of the loaf these molecules also influence the
flavour of the bread.

4. Enzymes important for bread manufacture.

The activity of amylase has been referred to above. Thus amylase is required to
hydrolyze starch to provide glucose for yeast fermentation: it is the latter which produces
the CO2 necessary for expansion of the bread dough i.e. for rising to occur. Thus the
endogenous glycolytic enzymes of yeast are required for oxidation of glucose formed
from starch. The action of amylase to alter the structure of starch also improves the gas
retention by the dough. The amount of endogenous amylase activity in flour, particularly
α -amylase activity, is variable and depends upon the source and harvest conditions of
the grain used to prepare the flour, so that addition of extra α -amylase is frequently
made, either during the milling of the flour or in the initial mixing of components to
prepare the dough. Fungal amylase addition leads to a dough which has better handling
properties and which has a fine crumb structure. The fungal enzyme is inactivated after
10 min at 75ºC. Bacterial amylases may be cheaper than those from fungi but care has to
be taken if a bacterial amylase is added which is relatively heat stable as excess activity
of such an enzyme produces a sticky crumb in the loaf. The activity of amylase is also
required to provide the reducing sugar, i.e. glucose, to provide the Colour of the final loaf
of bread.

The action of proteases can improve elasticity and handling properties of doughs and
give bread with a good volume. Most grains used for flour production have low
endogenous protease activity as the protease action normally increases once germination
of the grain occurs. Addition of a fungal protease, prior to dough mixing, may be
beneficial for bread manufacture as this can soften gluten specifically and has a limited
activity. Over-activity of added bacterial proteases or the plant protease, papain can lead
to decomposition of bread structure through excessive protein hydrolysis. These
proteases may be added to high protein flour used for biscuit manufacture where a dough
which is easy to roll out and does not rise much is required. The choice of protease is
limited by the action of endogenous protease inhibitors in flour and the pH of the dough.
The latter is about pH 5.5-6.0 for wheat dough but may be a lower pH (4.2) for rye
dough. The latter may be prepared as sour dough. Acidification occurs through the
activity of lactic acid bacteria. The rye dough is leavened by it’s endogenous
microorganisms or by addition of a starter culture.

Rye flour produces a weak gluten structure which does not hold the gas produced during
fermentation very well so that the bread is more dependent upon the carbohydrate
component. As rye flour contains a high activity of endogenous amylase the excessive
activity of this enzyme degrading the starch can reduce water binding and swelling of
starch producing a loaf with a poor volume. The activity of the amylase may also
contribute to the dark colour of the crumb and surface of the loaf and the sweet taste.

In the rye dough the pentosans play a vital role in dough formation. Rye flour prepared
from rye obtained in a dry harvest has few enzymes which gives a tough dough and low
volume bread: addition of pentosanase can produce a less tough dough, improve volume,
give a softer crumb and better storage properties.

Addition of soya flour to wheat flour is carried out to increase the activity of
lipoxygenase. This appears to have two beneficial commercial effects. Firstly the enzyme
bleaches the natural flour pigments, the carotenoids so increasing the whiteness of the
resulting bread. Secondly the peroxidation of certain lipids alters the lipid protein
interaction within the bread dough to give an improved crumb structure, after baking.

Careful balancing of proportions of endogenous and added amylase, protease,

pentosanase and lipoxygenase activity can improve volume, crumb structure and shelf
life. Thus enzymes are important in the formation, the appearance, the flavour and quality
of every loaf of bread manufactured.

The first enzyme to be isolated was diastase(an amylase) from malt in 1833 and the
enzyme activity in soy flour was patented as a bleaching agent in 1934. Now there are
companies who produce enzyme preparations specifically for baking. Other enzymes
which may be used more in the future, include glucose oxidase and sulphydryl oxidase,
patented as a dough strengthening agent in 1989, or a combination of hemicellulase and
glucose oxidase patented as a bromate replacer, also in 1989. There is, however, different
legislation covering the use of enzymes in baking, in different countries. There are also
controversial issues with respect to labelling of products. Should added enzymes be
regarded as processing aids, and if their activity is destroyed during baking be omitted
from labelling or are they additives which need to be recorded on the labels? These issues
are being discussed but have not yet been resolved.

For a detailed discussion of enzymes in wheat and other cereals that have technological
importance the reader is referred to the Bibliography.

Questions 2.I

1) What class of enzymes do the following enzymes belong to?

a) Amylase
b) Protease
c) Lipoxygenase.

2) Which enzymes have to be added during the preparation of bread to release CO2 to
help the bread rise?

3) Why is the temperature sensitivity of bacterial amylases, which may be added

during bread preparation, of importance?