An enzyme is a protein catalyst that makes  chemical changes in biological systems. Various categories are used in baked goods, beverages, dairy, beer, glucose syrups, starch and other food products.1 In bakery systems, enzymes act as: Dough conditioners Fermentation enhancers Anti-staling agents This enables bakers to remove undesirable additives and make clean label baked goods. Origin Enzymes are naturally present in many living organisms such as animals, plants, bacteria and fungi. There, they participate in metabolic processes.  Also, they can be found in food materials such as cereal flours, fruits and vegetables

1. Enzyme
• An enzyme is a protein catalyst that makes chemical changes in biological systems.
Various categories are used in baked goods, beverages, dairy, beer, glucose syrups, starch
and other food products.1
In bakery systems, enzymes act as:
• Dough conditioners
• Fermentation enhancers
• Anti-staling agents
• This enables bakers to remove undesirable additives and make clean label baked goods.
Origin
• Enzymes are naturally present in many living organisms such as animals, plants, bacteria
and fungi.There, they participate in metabolic processes. Also, they can be found in food
materials such as cereal flours, fruits and vegetables

2. Function of Enzyme
• Similar to other enzymes, ones used in bakery products can facilitate chemical
reactions without undergoing any change in their molecular structure.
• They can be utilized continuously as long as they are not denatured. For
example, heat can stop their activity.Also, there needs to be enough substrate
in the flour.
• xamples of substrates are proteins, fats, sugars, starch or non-starch
polysaccharides (cellulose, glucans, arabinoxylans).
• Primary uses in bakery products
Fermentation aides
1. Mix reducers
2.Modifiers of dough handling properties
3. Dough strengtheners
4.Crumb softeners (anti-staling agents)

3. IUPAC/IUB classification Type
Chemical bond cleaved
Reaction catalysis/hydrolysis
Specific function
Hydrolases
Protease Protein – peptide bond
•Reduces mixing time
•Modifies dough handling properties
•Improves pan flow
•Improves dough machinability
•Redistributes water in dough
Lipase Fats – Ester (alcohol-fatty acid) bond
•Produces emulsifiers from fats
•Stabilizes gas cells in batter and dough
•Improves batter aeration
•Softens crumb, helping fight staling
Amylases Starch – glycosidic bond
•Improves fermentation
•Produces maltose and glucose from starch (yeast food)
•Anti-staling agents
Cellulase Cellulose- glycosidic bond •Redistributes water in dough
•Improves gluten matrix cohesiveness and gas retention
•Modifies dough consistency and handling properties
Xylanase / Pentosanase / Hemicellulase Arabinoxylans – glycosidic bond
Maltase, invertase Disaccharides, maltose and sucrose – glycosidic bond
•Improves fermentation
•Produces monosaccharides (yeast food)
•Improves crust color, through browning reactions
Asparaginase Proteins – amide (carbon-nitrogen) bond •Reduces acrylamide in thermally processed foods
Oxidoreductases
Glucose oxidase Glucose – oxidation into gluconic acid and hydrogen peroxide •Strengthens dough
•Aggregates gluten-forming proteins, through the oxidation of
sulfhydryl (–SH) groups to disulphide (S–S) bond
•Improves mixing tolerance
•Enhances dough gas retention capacity
Hexose oxidase
Oxidation of hexose sugars (less specific) by atmospheric
oxygen into gluconic acid and hydrogen peroxide
Lipoxygenase Fatty acids – oxidation to produce peroxides
•Similar functionality to that of glucose/hexose oxidase
•Bleaching effect (destruction of flour carotenoid pigments).
Transferases Transglutaminase
Acyl-transfer reaction between carboxamide group of
peptide-bound glutamine residues and a
variety of primary amines
•Similar functionality to glucose/hexose oxidase although
using different mechanism (introduction of covalent cross-
links between glutamine and lysine).

4. Application of Enzymes
• Bakery enzymes are micro ingredients usually added at levels of 0.005–0.01% (50–100 ppm based on flour
weight). How much depends on the formulation and process needs.They require special conditions for
optimum activity and performance:1,2
• Acidity (pH): Most bakery varieties perform well within the pH range of 4.0–7.5.
• Temperature: Chemical reaction rates double for every 18°F (10°C) increase in temperature. Optimal
temperature for most commercial ones is 95–140°F/35–60°C.
• Contact time between enzyme and substrate: Enzymes need time to act on the substrate.
• Aqueous dispersion medium to support chemical reactions.
• Amount of substrate: they work better when excess of substrate is available.
• Adequate enzyme dosage
FDA Regulation
• Enzymes used in the bakery industry are GRAS (Generally Recognized as Safe) food additives in the US.The
FDA regulates their source or origin (food-compatible) and establishes limits to their use (if applicable) based on
GMP.

5. Xylanase
(Arabinoxylan
s (both water-
extractable
and water-
unextractable)
Definition
Xylanase is one type of hydrolytic enzymes which targets
arabinoxylan fibers naturally present in cereal flours, both
refined and whole grain.
Origin
Xylanase can be obtained from fungal and bacterial
fermentations.
Function
It plays different roles that positively affect the gluten-
starch matrix integrity
Xylanase helps obtain the following characteristics:
• Increased pan flow
• Higher dough extensibility
• Increased dough machinability during makeup
• Superior gas retention for optimum oven spring and
product volume
• Stronger and more continuous gluten network to
minimize gas bubble coalescence
• Finer and more even gas cell structure in the crumb

6. Mechanisms used
by xylanase to
improve overall
product quality
• Hydrocolloid effect
• Oxidative gelation ofArabinaxylanase
interaction This is known as oxidative
gelation and creates larger and more
stable molecules known as
Arabinoxylanase gels which reinforce the
gluten network.
• Protein-and-Arabinaxylanase interaction
Xylanase promotes further interaction
between xylanase and gluten proteins to
reinforce the polymeric gluten network
and enhance oven spring.
• Dough relaxation effect
• Foam stabilization effect

7. AscorbicAcid
Ascorbic acid, also known as vitamin C, is an essential nutrient
found in citrus fruits and many vegetables. It is used as wheat
flour improver in yeast-leavened baked goods to help increase
the volume of bread and provide better tolerance to variable
processing conditions, such as dough temperatures and
proofing times.
ascorbic acid as a flour improver, or dough conditioner,At
doses as low as 20–30 mg per kilogram of flour bread volume
increased by 20%.¹,²
Oxidizing agents In Bread Processing
• Potassium bromate
• calcium peroxide
• ascorbic acid
Function
• One prominent function of ascorbic acid in bread dough is
to stabilize the gluten protein network.This is reflected in
greater loaf volume and a finer, more uniform crumb
structure.
• Ascorbic acid itself is a reducing agent with strong
antioxidant properties in food systems

8. Gluten
strengthenin
g Effect of
Ascorbic
Acid
• Higher gas retention capacity of
dough
• Greater elasticity of dough
• Higher dough tolerance to over-
proofing and over-mixing
• Higher water absorption of flour
to obtain equivalent dough
rheology after mixing
• Diminished dough viscous
behaviour
• Higher dough resistance to
deformation
• Enhanced oven spring during
baking

9. Application and
Factors that
affect the
oxidizing
potential of
ascorbic acid
Application
• Ascorbic acid is a white granular powder that is often used in
bread and buns at levels up to 150 ppm (based on flour
weight).The exact amount dosed of AA depends on the
following factors:
• Protein level of flour
• Breadmaking system used (e.g. sponge and dough, no-time
dough, Chorleywood bread process)
• Bake test results
• Scaling weight/pan volume ratio
• Amount of other oxidizing agents (e.g. ascorbic acid forADA
or bromate replacement)
• Amount of bran, whole grains and fruit inclusions in
formulation (whole wheat and multigrain bread require a
higher amounts of AA)
Factors that affect the oxidizing potential of ascorbic acid
• Amount and enzyme activity of ascorbate oxidase in flour
• Amount of atmospheric oxygen during mixing.

10. Maltase
Enzyme
• Maltase is one type of alpha-glucosidase
enzymes that splits disaccharides like maltose
into their constituent glucose units.
• Maltose itself cannot be used or metabolized
by baker’s yeast cells.
• However, when broken down into its glucose
components, the latter are quickly consumed
by yeast and used for cell growth and
metabolism (e.g., fermentation reactions in
the absence of oxygen).

11. Enzymes that can be produced
by baker’s yeast cells include:
•Invertase
•Maltase
•Zymase

12. What is lipase?
• Lipase is a clean label ingredient, considered as a processing
aid.
• It can partially or totally replace traditional dough
strengthening emulsifiers, such as anionic surfactants DATEM
and SSL.
• Lipase can also replace crumb softening products such as
glyceryl monostearate (GMS)
• It can be found in many plants and animals. In recent decades,
lipases have been produced commercially from highly
specialized microorganisms such as bacteria, yeast and fungi.
Lipase is one type of
hydrolytic enzymes
responsible for
degrading or modifying
nonpolar (e.g.
triglycerides) and polar
lipids (e.g. galactolipids
and phospholipids)
found in both animal
and plant sources.

13. Lipase Enzyme Ingredient replaced
Lipase
EC
number
Substrate Reaction catalyzed
Ingredient
replaced
Triacylglycerol
lipase
3.1.1.3
Non-polar lipids in vegetable fats
and oils
Triacylglycerol (TAG) + H2O →
Diacylglycerol (DAG) + Free fatty acid (FFA)
Diacylglycerol (DAG) + H2O →
Monoacylglycerol (MAG) + FFA
GMS
Distilled
monoglycerides
Phospholipase
A1
3.1.1.32
Polar lipids found in cereal flours
and soy/sunflower lecithin
Phosphatidyl-choline (PC) + H2O → Lysophosphatidyl-
choline (LPC) + FFA
Phosphatidyl-ethanolamine (PE) + H2O →
Lysophosphatidyl-ethanolamine (LPE) + FFA
N-acyl-phosphatidyl-ethanolamine (NAPE) + H2O → N-
acyl-lysophosphatidyl-ethanolamine (NALPE) + FFA
DATEM, SSL, CSL
Egg reduction /
egg replacement
Phospholipase
A2
3.1.1.4
Polar lipids found in cereal flours
and soy/sunflower lecithin
DATEM, SSL, CSL
Egg reduction /
egg replacement
Galactolipase 3.1.1.26
Polar lipids found in cereal flours
and soy/sunflower lecithin
Digalactosyl-diglyceride (DGDG) + H2O → Digalactosyl-
monoglyceride (DGMG) + FFA
DATEM, SSL, CSL

14. Lipoxygenase
Enzyme
• Lipoxygenase, also known as linoleate: oxidoreductase, is an
enzyme used in commercial food production.
• In bread making, it both bleaches flour and improves dough
handling.
Origin
The enzyme is found in plant and animal tissues, but it is most
abundant in beans, peas, and potatoes. For use in commercial baking,
enzyme-active soybean flour is typically used, although it can also be
found in fava bean and other bean flours. Roughly half of the
commercially-available enzymes on the market are sourced from
genetically-modified organisms.
Function
Enzymes are a key ingredient in most bakery products as their use
improves the final product’s properties.
Lipoxygenase is an oxidative improving agent commonly used in
doughs in lieu of potassiumbromate and azodicarbonamide or other
chemical improvers.
Application
• Typically, enzyme-active soybean flour, which contains
lipoxygenase, is added to a dough. No more than 0.5 percent is
used. Experts suggest judicious use to prevent off flavors from
being created as a result of its use.2
• In some applications, enzymes might be custom blended to take
fullest advantage of their collective, and often chemically
synergistic, properties

15. Protease
Enzyme
Difinitions
Protease is an enzyme that catalyzes the hydrolysis of proteins. It does this
by breaking down peptide bonds between amino acids.This enzyme is used
in bakery products as a dough conditioner. It modifies dough rheology and
handling properties, improving:
• Workability
• Pliability
• Machinability
• The quality of the finished products
Origin
• Protease is a natural biological catalyst. It can be extracted from cereals
and fruits, such as papain or bromelain. Or, it comes from animal
sources like chymosin or rennin from bovines.
Application
• Protease is called a micro ingredient in baking. It is favored in
breadmaking systems that don’t require fermentation steps such
as yeast preferments or sourdough.The addition of specialty enzymes is
how high-quality artisan and variety bread can be made in a short time.
• Usually, protease is added during dough mixing. Depending on the
application and formulation, levels are 0.1 to 0.5% based on flour
weight. It is also used in clean label applications.There can be issues with
purity, such as side reactions and secondary activity. Also, ensuring
complete inactivation during baking is very important for optimal
performance.When supplying protease, it is a good practice to conduct
baking tests and lab analysis prior to line production.

16. Function
• Protease performs different functions in bread and cracker dough
systems.They are classified by their action pattern:
• Endoprotease: can split the protein interior chains at random
linkages to form polypeptides, peptides and peptones.This results
in a reduction of the molecular size of proteins which greatly
impacts dough rheology. Endoprotease provides the following
functions:2,3
• Through the breakdown of proteins, the gluten network is
weakened.This enhances extensibility and decreases resistance to
deformation. Protease-treated doughs are easier to round, sheet
and mould.
• Reduce mixing energy and time in some breadmaking processes
that use no-time or straight dough systems.
• Improve the sheetability of crackers and pizzadoughs.
• Improve dough pan flow dough.
• Increase the bread crumb tenderness.
• Slightly increase dough stickiness.This is due to gluten breakdown
and water redistribution among flour (starch, arabinoxylans and
cellulose).
• Ease dough expansion during proofing and oven spring through
enhanced extensibility.This has a positive impact on the finished
product volume.

17. Hexose
Oxidase
Enzyme
• Hexose oxidase, or simply HOX, is a unique
bakery enzyme which can strengthen the gluten
network in yeast-leavened doughs and enhance
finished product volume. It can also contribute to
a ‘clean label’ claim.
• As an enzyme, HOX only functions as a means to
an end. Unlike glucose oxidase, HOX is fully
capable of hydrolyzing both mono- and di-
saccharides into sugar acids while at the same
time producing hydrogen peroxide as a reaction
byproduct (the true oxidizing agent).1
• It also functions as a mild dough drier to reduce
surface stickiness in high-speed flourless makeup
lines.
Origin
• Hexose oxidase (E.C. 1.1.3.5) enzyme can be
obtained, though unfeasibly, from carrageenan
seaweed, Chondrus crispus

18. Function
Hexose oxidase is an enzyme which, in the
presence of oxygen, is capable of oxidizing mono-
and di-saccharides, maltose and lactose to their
corresponding lactones or aldonic acids.
Glucose removal in egg processing for shelf-life
extension
Acrylamide reduction from Maillard reactions via
monosaccharides removal
Gluten network strengthening to increase volume
and improve proofed dough shock resistance

19. Laccase
Enzyme
• What is Laccase?
• Laccase is an enzyme used in baking to improve various
qualities of doughs and baked goods.The ingredient can be
used in bread production, for example, to enhance:
• Dough handling
• Loaf volume
• Crumb structure
• Origin
• Laccase is an ingredient sourced from fungi, which secrete
laccases outside their cells during fermentation.Only a few
types of fungi are used to produce laccase on an industrial
scale. 1 One used in baking is derived from the white-rot
fungus Trametes hirsute.2

20. Function
• Among the substances on which laccase acts are many compounds found in
foods, including carbohydrates and unsaturated fatty acids. laccase is specifically
used in baking to create crosslinks, permanent covalent bonds in foods that
contribute to food texture.
• Laccase is an oxidative enzyme, which forms links with proteins and select
carbohydrates, but can also act on lipids. It forms these crosslinks primarily with
arabinoxylan. In the process, it changes the rheology of gluten, dough, and bread.
These changes include improvements in dough handling, loaf volume, and crumb
structure in bread
Application
• In some experiments exploring and validating its addition to bread, 5 g of laccase
per 100 kg of flour was used.4
• Among the improvements reported to dough with the addition of laccase are
enhanced dough machinability due to increased dough strength and stability as
well as a reduction in stickiness.
• Breads featured better crumb structures and were softer.
• In doughs using an oat flour, the addition of a commercial laccase “significantly
improved” the properties of the dough and final bread product, which included
enhanced bread texture and volume.The crumb hardness and chewiness was
reduced

21. Amylase {Also
known as
amylolytic or
starch-
degrading
enzymes}
• Amylase is a hydrolytic enzyme that breaks
down starch into dextrins and sugars. It’s
made up of a family of starch-degrading
enzymes that include:1
• Alpha-amylase
• Beta-amylase
• Amyloglucosidase or glucoamylase
• Pullulanase
• Maltogenic amylase
• Amylases can work at the same time in
perfect synergy.They are key ingredients
that extend the shelf-life of bread, working
as fermentation improvers

22. Function
• Amylases perform the following functions in bakery products:
• Provide fermentable and reducing sugars.
• Accelerate yeast fermentation and boost gassing for optimum dough expansion during
proofing and baking
• Intensify flavors and crust color by enhancing Maillard browning and caramelization reactions.
• Reduce dough/batter viscosity during starch gelatinization in the oven.
• Extend oven rise/spring and improve product volume.
• Act as crumb softeners by inhibiting staling.
• Modify dough handling properties by reducing stickiness

23. Application
• Production of starch syrups
• Sprouting of cereals
• Anti-staling agents
• Alpha-amylase has the largest effect on dough properties and bread quality
• Provides food for yeast for optimum product volume and color
• Decreases proofing time in low‐sugar and frozen dough
• Production of high DE starch syrups
• Provides substrate for glucose oxidase to properly function as dough
strengthener