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1 AMYLASE ENZYME

1.1 Breadmaking

 Enzyme amylase is used in breadmaking to


increase volume, texture and flavor, and to
retard bread staling.

 Fungal amylase, which is usually α-amylase, is


added, because amylase degrades starch into
simple sugars (glucose, dextrin, maltose)

 The increased sugar content has several


benefits.
1.1 Breadmaking

 It provides enough carbohydrates for yeast


fermentation. More yeast fermentation increases
bread volume and produces better flavor.

 Less starch remaining means the chance for


starch recrystallization is decreased. Starch
recrystallization is known to be the main reason
for bread staling.

 So adding amylase prevents recrystallization and bread staling.


1.1 Breadmaking

 Bread crust color is created by Maillard reaction between simple reducing sugar
and protein. More simple sugar will result in faster color development.

Maltose Glucose

Enzyme
Glucose

Active site
1.2 Beer brewing

 The malting process develops enzymes that


reduce starches and proteins during malting
and mashing, which helps create better clarity,
head retention and body.

 Activity of two primary enzymes in beer:


• alpha-amylase
• and beta-amylase.
1.2 Beer brewing

 The alpha amylase used in the mash comes


from the malt, where it is entirely produced in
the aleurone layer during malting.

 Levels of alpha-amylase are typically high in


pale malt but are virtually zero in roasted malt
due to heat degradation.
1.2 Beer brewing

 Alpha-amylase breaks down large, complex,


insoluble starch molecules into smaller, soluble
molecules for the beta-amylase.

 It is stable in hot, watery mashes and will


convert starch to soluble sugars in a
temperature range from 145°F to 158°F.
1.2 Beer brewing

 These B-amylase helps create lighter bodies and


more alcohol and are most active from 131°F-149°F.
As the temperature approaches 149°F, these
enzymes are operating extremely fast, but are also
being denatured quickly.

 In short, if the mash is held at a temperature


within the beta-amylase range, then a greater
proportion of soluble sugars will be maltose and
thereby be more fermentable.
1.3 Alcohol producing

 People used to use enzyme amylase in malt,


but now they change to fungal amylase.

 In alcohol producing, amylase is used in


conversion process:
• Convert starch to dextrin by α-amylase
• Convert dextrin to fermentable sugar by
beta-amylase or amyloglucosidase (AMG)
1.3 Alcohol producing

 The basic role of alpha-amylase in alcohol


production is rapid translation at the cooking
stage and at the early stages of conversion,
dextrinization and sugar accumulation.
1.3 Alcohol producing

 The roles of amyloglucosidase:


• Hydrolysis of 1,4-glucoside in polysaccharide.
• They continuously cleave non-reducing glucose
radicals in the polysaccharide circuit.

 There is also the ability to cleave 1,6-glucoside


radicals.
 The final product produced is glucose.
1.4 Juice producing

 Amylase is used to decompose starch in fruit


during the internal filtration phase in juice
production.

 Fruits may have a high content of starch, when


processed from some apple varieties or unripe fruit.

 Food and specially fruit juices are complex systems that have Ca2+ ions and
probably Ca2+ sequestering agents as organic acids. These compounds can
affect the action of amylases used for processing and its thermal inactivation.
1.5 Fructose and Glucose syrup producing

 Starch is used in the production of fructose and glucose syrups.

 This process involves three steps:


 Gelatinization, Liquefaction, and Saccharification.

Gelatinization Involves the dissolving of starch granules in


water to form a viscous starch suspension.
The amylase and amylopectin are dispersed
into the water on dissolution.
1.5 Fructose and Glucose syrup producing

Liquefaction Liquefaction of starch is it’s partial hydrolysis into short


chain dextrins by α-Amylase resulting in reduction of
the viscosity of the starch suspension

Saccharification Saccharification is the production of glucose and


fructose syrup by further hydrolysis. This is carried out
by glucoamylase which acts as an exo-amylase by
cleaving the α-1, 4-glycosidic linkages from the non
reducing terminal.
1.5 Fructose and Glucose syrup producing

 The action of pullulanase along with glucoamylase yields high glucose syrup.

 This high glucose syrup can then be converted into high fructose syrup by

isomerization catalysed by glucose isomerase.

 The fructose syrup obtained is used as a sweetener, especially in the beverage

industry.
2 LIPASE ENZYME
2.1 Breadmaking

 Lipases and phospholipases are recognized as


an additional tool for improving bread-making
properties.

 Specific lipases are claimed to improve


dough-handling properties, increase strength
and stability.

 Besides this, such lipases also improve crumb


structure and crumb whiteness.
A. Applications in cake and muffin production

 Addition of commercial lipase to reduce the


surface tension and surface viscosity at the
air/water interface of batter.

 Result after baking was an increase in


cake specific volume and maintaining a fine
crumb structure.
B. Applications in pasta and noodle production

 Lipase addition extends shelf life and


keeps the characteristic yellow color over
time.

 Lipase makes boiled noodles firmer,


smoother and less sticky.
B. Applications in pasta and noodle production

 Polyphenol oxidase (PPO) cause problem


in darkening of raw noodles or noodle
dough sheets, during production or shelf
life.

 Treatment with fungal lipase helps to


brighten noodle sheets or raw noodles
during shelf life.
B. Application in pasta and noodle production

 Another common problem is speckiness:


that are small dark spots.

 By adding fungal lipase to the recipe,


the increase of the number of dark spots
over storage time can be highly reduced.
2.3 Dairy products making

Application in cheese production

 The traditional sources of lipases for cheese


flavour enhancement are animal tissues,
especially pancreatic glands (bovine and
porcine) and pre-gastric tissues of young
ruminants (kid, lamb and calf).
2.3 Dairy products making

Application in cheese production

 Enzyme modified cheese (EMC) is produced


when cheese is incubated in the presence of
enzymes at elevated temperature in order to
produce a concentrated flavour by lipase
catalysis such as dips, sauces, soups and
snacks.
2.3 Dairy products making

Cheese type Lipase source


Romano Kid/lamb pre-gastric
Domiati Mucor miehei
Camembert Penicillium camemberti
Mozzarella Calf/kid pre-gastric
Fontina Mucor miehei
Roquefort (Blue cheese) Penicillium roqueforti
Cheddar Aspergillus oryzae/ A. niger

Examples of lipase in cheese making and accelerated


cheese ripening
2.3 Dairy products making

 Fatty acid chemical derivatives released from milk fat by lipases.

 Lipolyzed Milk Fat (LMF): derived from short to medium chain fatty acids has
a creamy, buttery and cheesy aroma.

 Lipolyzed butter fat is a yellowish creamy


like liquid that has the odor of milk fat that
has undergone enzymolysis.
2.3 Dairy products making

 LMF can be applied in many popular products:

butter flavours for


artificial creams and
margarine
cheese flavour
chocolate coatings
syrups
additives.
2.4 Cocoa butter producting

 Cocoa butter (CB) contains palmitic and stearic acids and has a melting point of
approximately 37°C, leading to its melting in mouth which results in a cooling
sensation.
 Unilever filed a patent describing a mixed hydrolysis
and synthesis reaction to produce a cocoa butter
substitute using an immobilized lipase, Rhizomucor
miehei lipase, which carries out a transesterification
reaction replacing plamitic acid with stearic acid to
give the desired stearic–oleic–stearic triglyceride.
2.5 Fats and Oils Industry

 Lipases modify the properties of lipids by altering the location of fatty acid chains
in the glyceride and replacing one or more of these with new ones.

 Lipases catalyze the hydrolysis,


esterification and interesterification of oils
and fats.

 Lipase catalyzed interesterificaiton of fats and


oils to produce modified acylglycerols cannot
be obtained by conventional chemical
interesterification.
3 PROTEASE ENZYME
A. Application in Meat processing technology

 Papain, bromelain, pepsin, or bacteria proteases from Aspergillus, Bacillus, ... are
used as meat tenderizer to soften meat for cooking and increase the flavor for meat.

 Advantages:
• Vitamins are preserved
• Do not create byproducts
• Do not darken by hydrolysis
• Proteases increase protein digestibility, which
is beneficial for people with poor digestive
disease or hypersensitivity to protein.
B. Application in Soy sauce process

 In soy sauce process, they use Aspergillus oryzae protease to hydrolyze protein and
amylase to hydrolyze starch into sugar and amino acids.

Aspergillus
oryzae
C. Application in Dairy industry

 Naturally occurring proteases contribute


significantly to the flavor characteristics of cheese.

 They are used for the acceleration of cheese


ripening, to modify the functional properties and
reduce the allergenic properties of milk products

 Proteases are also used to hydrolyze the specific


peptide bond to generate paracasein and
macropeptides.
C. Application in Dairy industry

 Proteases are used to coagulate milk for the


production of cheese.

 Chymosin is added to milk to hydrolyze casein (a


milk protein), resulting in coagulation and “curding”.

 Chymosin is preferred because of its high


specificity for casein  excellent performance in
cheese making.
D. Application in Baking industry

 The protease from Aspergillus usamii has been


successfully employed for the improvement of
functional properties of wheat gluten.

 Proteases are widely used in baking industry


for the production of bread, baked foods,
crackers and waffles.
D. Application in Baking industry

 Advantages:

 Reduce mixing time  Regulate gluten strength in bread, to


control bread texture and to improve
 Assure dough uniformity
flavor
 Decrease dough consistency
 Frequently added to dough preparations
 Replace bisulfite while proteolysis  have great impact on dough rheology
breaks down peptide bonds. and the quality of bread due to effects
 weakening of the gluten network on the gluten network or on gliadin.
E. Application in Wine and beer industry

 Acidic fungal proteases are used in improving


fermentation of beer as they are efficient even at
low pH by balancing the amino acid profile of
beer.

 Protease increase soluble protein and free


amino – nitrogen (FAN)
 Malt improvement
 Improve yeast growth.