6.

Transformation of nutrients during processing and preservation

6.1. Browning reaction
In foods that are thermally processed, the Maillard reaction, or nonenzymatic browning,
between carbonyl and amino groups frequently occurs. The Maillard reaction is a desired side
effect of numerous commercial and home procedures, and it gives some cooked foods their
enticing flavor and brown color. The Maillard reaction has been known to cause the degradation of
various important amino acids, including lysine.
6.1.1. Enzymatic browning
The spontaneous occurrence of enzyme-induced browning is common in many fruits and
vegetables. When fruits and vegetables are bruised, cut, peeled, infected, or subjected to any
abnormal circumstances, brown melanins are produced by the oxidation of phenolic chemicals,
which causes the fruits and vegetables to darken quickly when exposed to air. Enzymatic browning
is caused by polyphenol oxidase (PPO), which is found in most fruits, vegetables, and some
seafood. Along with PPO, the related oxidative enzyme peroxidase such as catechol oxidase, and
other enzymes that create melanins and benzoquinone from natural phenols. may also cause fruit
and vegetable enzymatic browning. Polyphenol oxidase first converts phenols into quinones,
which are very susceptible to nucleophilic attacks from other proteins due to their strong
electrophilic state. Following a series of events, these quinones are polymerized, which finally
causes the development of brown pigments (melanosis) on the food's surface. The quantity of
active polyphenol oxidases in the food is a good indicator of the rate of enzymatic browning. As a
result, the majority of research into ways to stop enzymatic browning has focused on reducing
polyphenol oxidase activity. However, not every food browning has undesirable outcomes.

Fig 2. Example of a general reaction of polyphenols by polyphenol oxidase (PPO) that catalyzes
enzymatic browning. The production of quinones undergoes more reactions which eventually form
brown pigments on the surface of the food.

Enzymatic browning control has long been difficult for the food business. Food enzymatic
browning can be stopped or slowed down using a variety of techniques, each of which targets a
different stage of the chemical reaction. The various enzymatic browning control methods can be
divided into two main categories: physical and chemical. Usually, several techniques are applied.
Sulfites are potent anti-browning agents, but their usage has been put on hold because of the
potential risks associated with their use. Numerous studies have been done to determine the precise
forms of regulatory mechanisms that occur when the enzymatic process is encountered.
Controlling browning also entails taking steps to restore the color of the food after it has browned.
Physical methods
- Heat treatment

- Cold treatment
The most popular methods for preserving food and preventing deterioration are freezing and
refrigeration. In low temperatures, browning enzyme activity, or the pace of reaction, decreases.
Thus, refrigeration aids in preserving the original appearance, color, and flavor of fresh fruits and
vegetables. Fruits and vegetables are distributed and sold in stores while being refrigerated.
- Oxygen elimination
Eliminating oxygen from the surroundings slows down the browning response since the presence
of oxygen is essential for enzymatic browning. When food is preserved, air is removed or
substituted with other gases (like N2 or CO2), such as in modified atmosphere packaging, wine or
juice bottling, the use of impermeable films or edible coatings, or dipping into salt or sugar
solutions. This keeps the food out of direct contact with oxygen. Plastic or other impermeable
coatings keep food from evaporating moisture and being exposed to oxygen in the air. Packaging
materials infused with anti-oxidants, anti-microbials, and anti-fungal agents including butylated
hydroxytoluene (BHT) and butylated hydroxyanisole (BHA), tocopherols, hinokitiol, lysozyme,
nisin, natamycin, chitosan, and -polylysine are becoming more and more popular.
- Irradiation
Another way to increase food shelf life is to irradiate it using UV-C, gamma, x, and electron
beams. Ionizing radiation slows down the maturity and sprouting of produce that will keep for a
long time and kills the bacteria that cause food to spoil.
Chemical methods
- Acidification
Like other enzymes, browning enzymes are active within a narrow pH range. For instance, PPO is
hindered below pH 3 and exhibits its peak activity at pH 5-7. To maintain the desired pH in food
items, acidifying agents and acidity regulators are frequently utilized as food additives. Anti-
browning compounds include acidulants such citric acid, ascorbic acid, and glutathione. Numerous
of these substances also exhibit additional anti-browning properties, including chelating and
antioxidant activity.
- Antioxidants
In the food sector, antioxidants are frequently utilized as food additives. These substances interact
with oxygen and prevent the browning process from starting. Additionally, they prevent the
production of melanin and interfere with the byproducts of the subsequent reactions. Examples of
antioxidants with anti-browning characteristics include ascorbic acid, N-acetylcysteine, L-cysteine,
4-hexylresorcinol, erythorbic acid, cysteine hydrochloride, and glutathione.
- Chelating agents
Because copper is a necessary cofactor for polyphenol oxidase to operate, copper-chelating
substances prevent this enzyme from doing its job. Numerous substances with chelating action,
including citric acid, sorbic acid, polyphosphates, hinokitiol, kojic acid, EDTA, porphyrins,
polycarboxylic acids, and other proteins, have been investigated and employed in various sectors
of the food business. Some of these substances also have additional anti-browning properties, like
acidifying or antioxidant properties. Coating materials for food packaging use hinokitiol.

Other methods
- Natural agents
It is recognized that a variety of natural items and their extracts, including onion, pineapple,
lemon, and white wine, might prevent or delay the browning of specific goods. By preventing PPO
activity, onion and its extract demonstrate strong anti-browning characteristics. Apples and
bananas have been demonstrated to benefit from pineapple juice's anti-browning properties.
Lemon juice is added to doughs to give baked goods a brighter appearance. The anti-browning
characteristics of citric and ascorbic acids in lemon juice may help to explain this effect.
- Genetic modification
Arctic apples have been genetically modified to silence the expression of PPO, thereby delaying
the browning effect, and improving apple eating quality.1

6.1.2. Non – enzymatic browning

Caramelization
The process of caramelization, Non enzymatic browning is a chemical process that produces a
brown color in foods.
The non enzymatic browning is also caused by caramelization of sugars, interaction of quinines
with amines and amino acids.2
which involves browning sugar, is widely employed in cooking because of the sweet, nutty
flavor and brown color it produces. Three classes of polymers—caramelans (C24H36O18),
caramelens (C36H50O25), and caramelins (C125H188O80)—are responsible for creating the
brown hues. Diacetyl and other volatile compounds are generated throughout the process, giving
off the distinctive caramel flavor.
The non-enzymatic browning process of caramelization is similar to the Maillard reaction.
Caramelization is a pyrolytic reaction, not an amino acid-based one like the Maillard reaction. The
monosaccharides fructose and glucose are formed during caramelization of the disaccharide
sucrose.

Fig 3. Example caramelization of table sugar (sucrose) caramelizing to a brown nutty flavor
substance (furan and maltol)

The Maillard reaction

Louis Camille Maillard is credited with discovering the Maillard reaction, which is a
chemical reaction between an amino acid and a reducing sugar that frequently necessitates the
addition of heat and yields a range of aromas and scents. It develops in food and, depending on
factors such as temperature, time, humidity, pH, etc., it results in aromas and browning, which are
followed by adjustments to the organoleptic and nutritional properties of the food. The two
monosaccharides that participate most frequently are glucose and fructose, while animal products
typically contain ribose.
The Maillard reaction is a common reaction that occurs in foods that have been heated or that have
been kept at room temperature for an extended period of time. In goods like coffee, bread, and

1
"PPO silencing". Okanagan Specialty Fruits, Inc. 2019. Retrieved 14 November 2019.
"United States: GM non-browning Arctic apple expands into foodservice". Fresh Fruit Portal. 13 August 2019.
Retrieved 14 November 2019.
2
meat where a distinctive scent and a brown color are required, the development of the Maillard
reaction is encouraged. Contrarily, the food sector has made numerous attempts to minimize the
extension of the Maillard reaction in foods like milk and fruit juices, where brown colors signify a
subpar product. Additionally, the Maillard reaction is to blame for the formation of hazardous
substances and the reduction in nutritional content of many foods. Additionally, the Maillard
reaction and other widely occurring processes like lipid oxidation, caramelization, and ascorbic
acid degradation have distinct metabolic pathways. Therefore, from a technological standpoint,
fine-tuning the Maillard reaction alone is challenging. Currently, gaining novel foods with
healthier and more flavorful qualities depends on our understanding of the shared chemical
pathways of these processes.
Biochemistry of the early maillard reaction
The initial Maillaed reaction is the condensation of the free aldehyde group of carbohydrate
with either the e – amino group of lysine or hydroxylysine residues or the α – amino group of N –
terminal amino acid of protein. Only open forms of sugars react with proteins, the carbonyl group
of an acylic monosaccharide attaching to a protein amino group via nucleophilic attack to form a
labile aldimine ( Schiff base ). This product may hydrolyse back to glucose and protein or undergo
an Amadori rearrangement to form a 1-amino-1-deoxyfructose ( fructosamine), derivative by a
stable, though slightly reversible, ketoamine linkage (Fig. 1). This product can cyclise to a ring
structure ( N-substituted-1-amino-dexokytetopyranose). The rate of the Amadori rearrangement is
approximately one-sixtieth that of the dissociation to glucose and protein and also varies between
proteins.3

Fig. 1. The early Maillard reaction

Biochemistry of the late Maillard reaction

The formation of the Amadori product discussed above is considered the first step in the
Maillard or browning reaction, which was first observed in 1912 when it was noted that amino
acids heated in the presence of reducing sugars develop a yellow-brown colour. The Amadori
product can undergo a further series of dehydration and rearrangement reactions. It may
spontaneously fragment, releasing reactive ketoaldehyde dicarbonyl compounds
(deoxyglucosones) of which 3-deoxyglucosone is the most important. Deoxyglucosones further
react with amino groups of proteins to form intra-protein cross-links known as advanced glycation
end (AGE) products. AGE products can probably also form by condensation of two Amadori
products, or by reaction of deoxyglucosones with the Amadori product. AGE products are
3
irreversible and accumulate over the lifetime of the protein. Most AGE products are fluorescent
and to date the majority of clinical data have been generated by the measurement of protein –
linked fluorescence.

6.2. Biochemical process for food flavouring

The demand for natural tastes and aromas in food additives has steadily increased over the
past few decades. In-depth study into the use of biocatalysts for the production of flavor and
fragrance compounds has been done in response to the increased demand for natural flavors. By
employing microbial biocatalysts made from free fatty acids or other substrates, the
biotransformation process produces natural tastes and fragrances in one direction. Consumer
preference for natural flavor and fragrance manufacturing techniques is undeniable, which directs
the market's attention primarily toward the food industry. Natural tastes are substantially more
expensive than their chemically manufactured analogues, therefore this demand cannot be met by
isolating and extracting them from natural resources. One of the common methods is the
employment of biocatalysis with the proper precursors to increase the availability of natural
flavors and smells. The technology of biocatalysis for the synthesis or resolution of optically active
intermediates was established and extensively used by the early 2000s. Since then, biocatalysis has
developed into a microbial enzyme technology that is frequently used in enterprises for the
manufacture of valuable compounds for commerce. Great catalytic activity can be achieved by
using whole cells, cell lysates, or pure enzymes to convert a substrate into a substance known as a
bio-catalyst. However, its dependability and expense are the key barriers to its widespread use.
Process flavor is a food flavour created by heating different precursors together while
regulating the reaction's temperature, pH, and moisture content. The Maillard reaction is the
fundamental theory underlying flavor processing. This reaction is crucial for taste creation in food
processing, in addition to browning, nutritional value decrease, toxicity, and antioxidant qualities.

6.1.2. Non – enzymatic browning

Reference:
Browning: Enzymatic Browning
Y. Jiang, ... S. Zheng, in Encyclopedia of Food and Health, 2016 -
https://www.sciencedirect.com/topics/food-science/enzymatic-
browning#:~:text=Polyphenol%20oxidase%20(PPO)%20present
%20in,browning%20of%20fruits%20and%20vegetables.

Handbook of Food Science and Technology 1: Food Alteration and

Food Quality
Chapter 5

Non-Enzymatic Browning
Thomas Croguennec
Book Editor(s):Romain Jeantet, Thomas Croguennec, Pierre Schuck, Gérard Brulé
First published: 08 January 2016
https://doi.org/10.1002/9781119268659.ch5
Citations: 5

https://www.slideshare.net/omar-alajil/non-enzymic-browning

Maillard Reaction
J.A. Rufián-Henares, S. Pastoriza, in Encyclopedia of Food and Health, 2016

https://www.sciencedirect.com/science/article/abs/pii/S0308814623018356

https://www.sciencedirect.com/science/article/abs/pii/S0308814623018356

BẢN TÓM TẮT:

6. Transformation of nutrients during processing and preservation
6.1. Browning reaction
In foods that are thermally processed, the Maillard reaction, or nonenzymatic browning,
between carbonyl and amino groups frequently occurs. The Maillard reaction is a desired side
effect of numerous commercial and home procedures, and it gives some cooked foods their
enticing flavor and brown color. The Maillard reaction has been known to cause the degradation of
various important amino acids, including lysine.
6.1.1. Enzymatic browning
− Enzymic browning is an oxidation reaction that takes place in some foods, mostly fruit
and vegetables, causing the food to turn brown.
− Even though enzymatic browning is not harmful, nor does it affect flavor, it’s just not a very
appetizing appearance.
− The enzyme catalyzing enzymatic browning in fruits is polyphenol oxidase or phenolase. In
chemistry, there’s a class of phenolic compounds, that are present in fruits and vegetables,
known as polyphenols, which form the substrate for this browning enzyme. Polyphenol
oxidase is found in plastids and chloroplasts.
− Polyphenols can be classified into different categories like anthocyanins, flavonoids, and
non-flavonoids. Anthocyanins are pigments present in fruits, flavonoids are occur in
catechins, tannins, and beverages, and non-flavonoids are a component of gallic acid in tea
leaves.
− These polyphenols are unstable and undergo oxidation when exposed to air through a series
of biochemical reactions involving conversion of one substrate to a product. The most
common chemical reaction taking place is the catalysis of first step of biochemical reactions
involving formation of quinones from phenolic compounds, and subsequent polymerization
of quinones to insoluble polymers known as melanin. This compound called melanin is
responsible for the browning of fruits and vegetables.
 Schematic Diagram of Enzymatic Browning

Enzymatic browning can be prevented by application of several methods. These methods are
mostly targeted to inactivate the enzyme and remove oxygen from the product.
Physical methods
- Heat treatment
o Heat treatment is the technique that is most frequently employed to stabilize foods.
o The most popular technique for inactivating vegetable enzymes is blanching.
o Because it results in losses of vitamins, flavor, color, texture, carbs, and other water-
soluble components, it is rarely used for soft fruits.
o It induces denaturation and subsequently inactivation of the enzymes but also causes
the destruction of thermosensitive nutrients. By exposing the food item to high
temperatures for long enough to denature the pro
- Cold treatment
o The most popular methods for preserving food and preventing deterioration are
freezing and refrigeration.
o In low temperatures, browning enzyme activity, or the pace of reaction, decreases.
- Oxygen elimination
o Eliminating oxygen from the surroundings slows down the browning response since
the presence of oxygen is essential for enzymatic browning. When food is preserved,
air is removed or substituted with other gases (like N2 or CO2), such as in modified
atmosphere packaging, wine or juice bottling, the use of impermeable films or edible
coatings, or dipping into salt or sugar solutions.
o Plastic or other impermeable coatings keep food from evaporating moisture and
being exposed to oxygen in the air.
o Packaging materials infused with anti-oxidants, anti-microbials, and anti-fungal
agents including butylated hydroxytoluene (BHT) and butylated hydroxyanisole
(BHA), tocopherols, hinokitiol, lysozyme, nisin, natamycin, chitosan, and -
polylysine are becoming more and more popular.
- Irradiation
Another way to increase food shelf life is to irradiate it using UV-C, gamma, x, and electron
beams. Ionizing radiation slows down the maturity and sprouting of produce that will keep for a
long time and kills the bacteria that cause food to spoil.
Chemical methods
- Acidification
o Like other enzymes, browning enzymes are active within a narrow pH range.
o To maintain the desired pH in food items, acidifying agents and acidity regulators are
frequently utilized as food additives.
o Anti-browning compounds include acidulants such citric acid, ascorbic acid, and
glutathione. Numerous of these substances also exhibit additional anti-browning
properties, including chelating and antioxidant activity.
- Antioxidants
In the food sector, antioxidants are frequently utilized as food additives. These substances interact
with oxygen and prevent the browning process from starting. Additionally, they prevent the
production of melanin and interfere with the byproducts of the subsequent reactions.
- Chelating agents
o Because copper is a necessary cofactor for polyphenol oxidase to operate, copper-
chelating substances prevent this enzyme from doing its job.
o Numerous substances with chelating action, including citric acid, sorbic acid,
polyphosphates, hinokitiol, kojic acid, EDTA, porphyrins, polycarboxylic acids, and
other proteins, have been investigated and employed in various sectors of the food
business.
o Coating materials for food packaging use hinokitiol.

Other methods
- Natural agents
o It is recognized that a variety of natural items and their extracts, including onion,
pineapple, lemon, and white wine, might prevent or delay the browning of specific
goods.
o By preventing PPO activity, onion and its extract demonstrate strong anti-browning
characteristics.
- Genetic modification
Arctic apples have been genetically modified to silence the expression of PPO, thereby delaying
the browning effect, and improving apple eating quality.4

6.1.2. Non – enzymatic browning

Non enzymatic browning is a chemical process that produces a brown color in foods. The non
enzymatic browning is also caused by caramelization of sugars, interaction of quinones with
amines and amino acids.
Caramelization
Caramelization is a complex group of reactions that take place when sugars, either dry or in
concentrated aqueous solutions, are subjected to high temperatures to form unique brown colors
and flavors in baked products and other food systems.
Similar to Maillard reactions, caramelization is a distinct non-enzymatic browning process.
However, unlike Maillard reactions which require the availability of sugars and free amino groups,
caramelization takes place using only sugars as substrates. Both reactions can take place
simultaneously and are dependent on temperature, water activity and pH
4
"PPO silencing". Okanagan Specialty Fruits, Inc. 2019. Retrieved 14 November 2019.
"United States: GM non-browning Arctic apple expands into foodservice". Fresh Fruit Portal. 13 August 2019.
Retrieved 14 November 2019.
Fig 2. Example caramelization of table sugar (sucrose) caramelizing to a brown nutty flavor
substance (furan and maltol)