LECTURE NOTE: BCM 317 (FOOD BIOCHEMISTRY)

STARCH GELATINIZATION, ENZYMATIC AND NON-ENZYMATIC

BROWNING, NUTRIENT-NUTRIENT INTERACTION DURING HEAT
PROCESSING OF FOOD.

Thermal Processing of food

Thermal processing is defined as the combination of temperature and time required to

eliminate a desired number of microorganisms from a food product. The basic purpose for
the thermal processing of foods is to reduce or destroy microbial activity, reduce or destroy
enzyme activity, and to produce physical or chemical changes to make the food meet a
certain quality standard.

STARCH GELATINIZATION

It is a process of breaking down the intermolecular bonds of starch molecules in the presence
of water and heat, allowing the hydrogen bonding sites (the hydroxyl hydrogen and oxygen)
to engage more water. This irreversibly dissolves the starch granule in water causing it to
swell. Water acts as a plasticizer or dispersants i.e additives that increase the deformation of
material undergoing non-reversible changes of shape in response

Three main processes happen to the starch granule: (1) granule swelling (2) crystal or double
helical melting, and amylose leaching.

Granule Swelling: During heating, water is first absorbed in the amorphous space of starch,
which leads to a swelling phenomenon.

Crystal or double helical melting: Water then enters via amorphous regions the tightly bound
areas of double helical structures of amylopectin. At ambient temperatures these crystalline
regions do not allow water to enter. Heat causes such regions to become diffuse, the amylose
chains begin to dissolve to separate into an amorphous form and the number and size of
crystalline regions decreases. Under the microscope in polarized light starch loses its
birefringence and its extinction cross.

Amylose leaching: Penetration of water thus increases the randomness in the starch granule
structure, and causes swelling; eventually amylose molecules leach into the surrounding
water and the granule structure disintegrates.

This gives the system a viscous and transparent texture. The result of the reaction is a gel,
which is used in sauces, puddings, creams, and other food products, providing a pleasing
texture. Starch-based gels are thermo-irreversible, meaning that they do not melt upon
heating Excessive heating, however, may cause evaporation of the water and shrinkage of the
gel.

The most common examples of starch gelatinization are found in sauce and pasta
preparations and baked goods.

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 In sauces, starches are added to liquids, usually while heating. The starch will absorb
liquid and swell, resulting in the liquid becoming thicker. The type of starch determines the
final product. Some starches will remain cloudy when cooked; others will remain clear.

Pasta is made mostly of semolina wheat (durum wheat flour), which contains high
amounts of starch. When pasta is cooked in boiling water, the starch in the pasta swells as it
absorbs water, and as a result the texture of the pasta softens.

Starch molecules make up the majority of most baked goods, so starch is an important part of
the structure. Starches can be fairly straightforward extracts of plants, such as cornstarch,
tapioca, or arrowroot, but there are also modified starches and pre-gelatinized starches
available that have specific uses.

List of different thickening and binding agents and their characteristics.

Cornstarch

Preparation: (1) A slurry (mixture of cornstarch and water) is mixed and added to a
simmering liquid while whisking until it dissolves and the liquid thickens

(2) Mixture of cornstarch with sugar and cold liquid, then the thickened mixture is simmered
until no starch taste remains

Uses and Characteristics: Used to thicken sauces when a clear glossy texture is desired, such
as dessert sauces and in Asian-inspired dishes

Translucent, thickens further as it cools; forms a sliceable gel

Sensitive to extended heat exposure, so products become thin if held at heat for long periods
of time

Waxy maize and waxy rice

Preparation: It is added to hot liquid while whisking until it dissolves and the liquid
thickens

Uses and characteristics: Used in desserts and dessert sauces

Clear, does not thicken further as it cools

Does not gel at cool temperatures, good for cold sauces

Quite stable at extreme temperatures (heat and freezing)

Modified starches

Modified starches, also called starch derivatives are starch prepared by physically,
enzymatically, or chemically treating native starch to change its properties. Modified starches

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are used in practically all starch applications, such as in food products as a thickening agent,
stabilizer or emulsifier; in pharmaceuticals as a disintegrant; or as binder in coated paper.
They are also used in many other applications. Starches may be modified to increase their
stability against excessive heat, acid, shear, time, cooling, or freezing; to change their texture;
to decrease or increase their viscosity; to lengthen or shorten gelatinization time; or to
increase their visco-stability.

Preparation: Added to hot liquid while whisking until it dissolves and the liquid thickens

Uses and Characteristics: Modified starches are often used in commercially processed
foods and convenience products

Modified to improve specific characteristics (e.g., stability or texture under extreme

conditions; heat and freezing)

Translucent, thickens further as it cools

Pre-gelatinized Starches

Pre-gelatinized starch is a starch that has been pre-cooked, dried and ground into flake or
powder form. Quickly and easily dissolving in cold liquids, pregelatinized starches allow
products to develop full viscosity without cooking.

Preparation: Added to liquid at any temperature

Uses and Characteristics: Used when thickening liquids that might lose colour or flavour
during cooking

Become viscous without the need for additional cooking, translucent, fairly clear, shiny, does
not gel when cold

Arrow root

Preparation: It is added to hot liquid while whisking until it dissolves and the liquid
thickens

Uses and Characteristics: It is derived from cassava root, used in asian cuisines , very clear
and possess a gooey texture . Translucent , shiny , very light gel when cold

NON-STARCHY AND THERMO-REVERSIBLE GELS

Gelatin

Preparation: It is usually powder or sheets, dissolved in cold water , added simmering

liquid ,activate with heat and sets when cold

Uses and Preparation: is a water-soluble protein extracted from animal tissue and used as a
gelling agent, a thickener, an emulsifier, a whipping agent, a stabilizer, and a substance that
imparts a smooth mouth feel to foods. It is thermo-reversible. Gelatin is available in two
forms; powder and sheet (leaf).

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Gelatin is often used to stabilize whipped cream and mousses; confectionery, such as gummy
bears and marshmallows; desserts including pannacotta; commercial products like
margarines; and dairy products such as yogurt and ice cream. Gelatin is also used in hard and
soft gel capsules for the pharmaceutical industry.

Agar agar

Preparation: It is usually powder, dissolved in cold water , added simmering liquid ,activate
with heat and sets when cold

Uses and characteristics: it is an extract from red algae and is often used to stabilize
emulsions or foams and to thicken or gel liquids. It is thermo-reversible and heat resistant.

It is typically hydrated in boiling liquids and is stable across a wide range of acidity levels. It
begins to gel once it cools to around 40ºC and will not melt until it reaches 85ºC .

FACTORS THAT AFFECT GELATINIZATION OF STARCH ( Assignment)

RETROGRADATION

Gelatinized starch, when cooled for a long period (hours or days), will thicken and rearrange
itself again to a more crystalline structure; this process is called retrogradation. During
cooling, starch molecules gradually aggregate to form a gel. The following molecular
associations can occur: amylose-amylose, amylose-amylopectin, and amylopectin-
amylopectin..

Due to strong associations of hydrogen bonding, longer amylose molecules (and starch which
has a higher amylose content) will form a stiff gel. Amylopectin molecules with longer
branched structure (which makes them more similar to amylose), increases the tendency to
form strong gelswhile high amylopectin starches will have a stable gel. Retrogradation
restricts the availability for amylase hydrolysis to occur which reduces the digestibility of the
starch.

Example of retrogradation process:

In baking, the starch present in bread doughs and batters becomes gelatinized. During this
process the starch goes from an ordered ,crystalline state to a disordered ,amorphous state.
Upon cooling the disordered starch state begins to retrograde or re-order , returning the starch
back to its more rigid crystalline state, resulting in the firming of crumb texture in baked
goods. Starch retrogradation is a time and temperature dependent process. When the starch
stays as a gel , a product is softer and we say is softer and we say it is fresh , When the starch
regains its crystalline form, , the product becomes firmer, and we say it is stale, the technical
term for this is starch retrogradation

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BROWNING

Browning is a common colour change seen in food during preparation, processing, or storage
of food. It occurs in varying degrees in some food material. The colour produced range from
cream or pale yellow to dark brown or black, depending on the food item and the extent of
the reaction.

Browning reactions may be desirable or undesirable. In some food, the brown colour and
flavour developed during browning is highly desirable and associated with a delicious, highly
acceptable, and quality product. Browning reactions contribute to the aroma, flavour and
colour of the product such as the browning crust of bread, and all baked goods, potato chips,
roasted nuts, roasted coffee beans and many other processed foods.

The undesirable effects of browning reactions are seen in dehydrated food such as eggs, dry
fruits, in cut fruits and citrus fruit juice concentrates, in canned milk and in coconut.

The colour varies from light cream to black while coconut develops a saffron colour. The off-
colour and off odour developed in foods depends on the extent to which the browning
reaction has progressed. Off flavours may vary from mild flavour changes to stale and very
bitter.

Controlled browning is necessary even in foods where browning is desired because excessive
browning can produce an undesirable product.

TYPES OF BROWNING

ENZYMATIC AND NON- ENZYMATIC BROWNING

ENZYMATIC BROWING

Enzymatic browning occurs when the enzyme polyphenol oxidase or other enzymes catalyze
the oxidation of phenols in the fruit to form compounds called quinones. The quinones can
then polymerize to form melanins, which cause the brown pigments.

It can be observed mainly in fruits (apples, bananas, grape, coconut ), Vegetables (potatoes,
mushrooms, lettuce) and some seafoods, is usually not desirable and creates heavy economic
losses for growers.

Enzymatic browning is detrimental to quality, particularly in post harvest of fresh

fruits ,juices and some shellfish. It may be 50 percent responsible of all losses during fruit
and vegetable production .on the other hand enzymatic browning is essential for the color and
taste of tea, coffee and chocolate

Importants points to note in enzymatic browning

 Light colour Fruits and vegetables darken when exposed to air as a result of the
presence of oxidative enzymes.
 Enzymatic browning occurs in those fruits and vegetables when the cellular
organization is disrupted by cutting, bruising or other injuries to the tissues.

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  This is due to the action of oxidative enzymes on the presence of phenolic substances
present in the fruit and vegetable tissues.

 Enzymatic browning takes place only in fruits and vegetables which contain phenolic
compounds. These phenolic compounds act as the substrate and in the presence of
oxygen and by the action of enzymes. These phenolic compounds are responsible for
the colour of many plants

NON-ENZYMATIC BROWNING

Non enzymatic browning is a chemical process that produces a brown colour in foods without
the activity of enzymes. Melanin and other chemicals are responsible for the brown colour.
There are two types of non enzymatic browning:

Maillard Reaction

Maillard reaction is a chemical reaction in which carbonyl group of a reducing sugars, react
with naturally occurring nitrogen –containing compounds such as amine group of amino acid
form a brown pigment known as melanoidins. The maillard reaction is a between an amino
acid and a reducing sugar, usually requiring the addition of heat. The reactive carbonyl
group of the sugar interacts with the nucleophilic amino group of the amino acid, and poorly
characterized odour and flavour molecules result. This process accelerates in an alkaline
environment because the amino groups do not neutralize. This reaction is the basis of the
flavouring industry, since the type of amino acid determines the resulting flavour. In the
process, hundreds of different flavour compounds are created. These compounds in turn
break down to form yet more new flavour compounds, and so on. Each type of food has a
very distinctive set of flavour compounds that are formed during the maillard reaction. It is
these same compounds that flavour scientists have used over the years to create artificial
flavours.

Melanoidins are brown, high molecular weight heterogeneous polymers that are formed
when sugars and amino acids combine through the Maillard reaction at high temperatures and
low water activity. Melanoidins are commonly present in foods that have undergone some
form of non-enzymatic browning, such as barley malts , bread crust, bakery products and
coffee. They are also present in the wastewater of sugar refineries, necessitating treatment in
order to avoid contamination around the outflow of these refine

This reaction occurs when virtually all foods are heated and also occurs during storage, most
of the effect of maillard reaction including the caramel aromas and golden brown colour are
desirable, Nevertheless , some of the effect of maillard reaction including foods darkness and
off-flavour development which are not desirable.

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CONTROL FACTORS THAT AFFECT MAILLARD REACTION ( Read up)

Ph, Temperature and Water activity

Caramelization

is a non-enzymatic reaction that occurs when carbohydrates or sugars in food are heated. It is
the process of removal of water from a sugar followed by isomerization and polymerisation
steps. When sugar is heated at high temperature (160 deg C) in the absence of water and
amino acids, it turns brown and this sugar-sugar interaction is known as caramelization.
Sugar is broken down into a number of compounds due to intense heat.

During a caramelization reaction , the sugars initially undergo dehydration and

polymerisation into complex molecules of varying molecular weights. Lightly coloured,
pleasant –tasting caramel flavours are produced during initial stages but as the reaction
continues higher molecular weight color bodies are produced and the flavour characteristics
become more bitter. Development of caramel flavour and colour in caramel custard is a
desirable change, whereas uncontrolled heating of sugar can lead to objectionable burnt
colour and flavour.

CONTROL OF BROWNING

1. Blanching

Blanching (scalding vegetables in boiling water or steam for a short time) is a must for almost
all vegetables to be frozen. It stops enzyme actions which can cause loss of flavor, color and
texture.

Blanching cleanses the surface of dirt and organisms, brightens the color and helps retard loss
of vitamins. It also wilts or softens vegetables and makes them easier to pack.

Blanching time is crucial and varies with the vegetable and size. Under blanching stimulates
the activity of enzymes and is worse than no blanching. Over blanching causes loss of flavor,
color, vitamins and minerals.

2. Temperature

Low temperatures can also prevent enzymatic browning by reducing rate of reaction.

3. Treatment with antioxidant agents

Antioxidants can prevent the initiation of browning by reacting with oxygen. They also react
with the intermediate products, thus breaking the chain reaction and preventing the formation
of melanin. Their effectiveness depends on environmental factors such as pH, water activity
(aw), temperature, light and composition of the atmosphere. The main antioxidants
hexylresorcinol , erythorbic acid, N-acetyl cysteine , cysteine hydrochloride , ascorbic acid
and glutathione

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[Link] with chelating agents

Polyphenol oxidase(PPO) requires copper ions to be active . Thus, the presence of a

substance capable of binding divalent cations present in the medium reduces the enzymatic
activity of PPO. The principal chelating agents are kojic acid, citric acid (Usually citric acid
is used for its chelating role, but also for acidifying the medium.)
Proteins can exert an inhibitory effect on PPO activity by Chelating the essential copper at
the active site of PPO through competitive inhibition, inhibiting its activity

5 Ion-Exchange Filtration

During wine synthesis, the use of Ion-exchange filtration is used to remove the brown color
sediments in the solution.

6 Freezing

Freezing is a technique often used to stop browning reactions in fruit. Freezing causes a
decrease in available water for enzymatic reactions. It is confirmed by Lavelli (2010) who
found that in the apple a water activity below of 0.3, the PPO is no longer active

7 Treatment with agents of firmness

Calcium salts are the best known; they are used in the strengthening of cell walls. The cell
walls are more stable to different treatments. This prevents the destruction of cell
compartments and also the contact of PPO with polyphenols in the [Link] main agents
of firmness are calcium lactate , calcium propionate , calcium chloride , calcium ascorbate
and sodium chloride

8 Treatment with acidifying agents

PPO is sensitive to pH variations. The fruit is a naturally acidic environment, additional

acidification may reduce the PPO activity or inactivate it below pH 3. The main acidifying
agents are citric acid like in lemon , erythorbic acid , ascorbic acid and glutathione.

9 Conservation in modified atmosphere

Oxygen is essential for the oxidation reaction and PPO activity, a solution to control
enzymatic browning reactions would be to change the oxygen content of the storage
atmosphere .Modifying the composition of atmosphere, showed that the enzymatic systems
are delayed without altering product quality

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NUTRIENT-NUTRIENT INTERACTION

Nutrient-nutrient interaction means the impact of the nutrient on other nutrient’s

bioavailability in mode of preparation of diet, in the intestinal lumen and in the
postabsorptive stage. Nutrient bioavailability includes two important components which are
absorption and utilization. Absorption is the process by which a nutrient moves from the
intestinal lumen into the body while utilization of the absorbed nutrients includes transport to
various parts of the body, assimilation by cells, and conversion to biologically active forms.

Nutrient-nutrient interactions may affect bioavailability either positively or negatively: may

either enhance or inhibit nutrient absorption or utilization, High or low levels of one or more
nutrients may affect bioavailability of other nutrient. Interaction can affect all the major
categories of nutrients; protein, carbohydrates, fats, vitamins and minerals. Interactions of
nutrients with non-nutrient components of foods can also alter availability.

Nutrient-nutrient interaction can be unidirectional or bidirectional interactions; unidirectional

interaction where one nutrient affect the biological disposition of another and bidirectional
interactions which usually occur among nutrients with similar physicochemical properties
and a common mechanism of absorption or metabolism.

Nutrient interactions can occur at several different levels:

In the diet: The mode of preparation of diets may be as important as diet composition
in determining nutrient interactions. For example, cooking in an alkaline medium may
decrease the interaction between ascorbic acid and iron by destroying the vitamin.

In the intestinal lumen: Interactions at this level determine the true availability of a
nutrient for translocation through the enterocyte( intestinal absorptive cells). Most
luminal interactions consist of direct nutrient-nutrient ones, but certain nutrients can
indirectly affect the absorption of others by modifying gastrointestinal physiologic
activities. For example, certain dietary fibres can stimulate gastrointestinal hormone
secretion or inhibit micellar formation, thus indirectly affecting nutrient absorption .

In the postabsorptive phase: Many interactions take place after the process of
absorption has been completed. These interactions may be in the form of
physiological synergism, such as that of vitamin A and zinc on the visual process, or
between vitamin A and iron mobilization. Conversely, negative interactions may
affect circulating or storage levels of nutrient.

Effect of Nutrient-Nutrient Interaction using Millard reaction as an example

The major chemical reactions that take place during food processing occur between the main
food components; the carbohydrates, proteins, fats and vitamins. These components can react
with each other and with various food additives such as nitrites, sulphite, aldehydes and
alkali: to give food products of lower nutritional value; to produce desirable and
undesirable browning and flavours; and very occasionally to produce toxic materials.

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 Proteins are the most reactive of the food components and they can combine with sugars,
fats, polyphenols, vitamin B6 and with various chemical additives such as nitrites, aldehydes
and alkali. These reactions can lead to a reduction in nutritional value, browning and flavour
formation and very occasionally to toxicity. Example: During early maillard reactions
reducing sugars combine primarily with the essential amino acid lysine to form a biologically
unavailable deoxyketosyl compound; this reaction is very important in milk products, which
can lose a large proportion of their lysine content during processing and storage without any
visible changes in colour. The later maillard reactions lead to the brown pigments; they
further reduce the nutritional value but also produce many desirable flavours and odours,
which have been reported to be at least partly responsible for the flavours and odours of
freshly baked bread, coffee, cocoa, beer, cooked meats and cooked vegetables.

Dietary interactions affecting nutrient bioavailability

Interacting Reported effects

nutrients
VITAMIN A
protein Protein deficiency decreases intestinal
absorption of vitamin A.
Diets with low (< 10%) or high
(20% -40% ) protein content inhibit
carotene dioxigenase activity.
Protein deficiency decreases the capacity to release
retinol from river stores.
Lysine-deficient proteins decrease
plasma retinol levels.
Protein quality affects the rhythm of
depletion of liver vitamin A reserves.
Corn- and black bean-based diets
cause lower rate of depletion than
casein, even when they provide more
protein.
fat An increase in dietary fat content
increases carotene absorption.
Polyunsaturated fats inhibit carotene
absorption and metabolism.
vitamin E Supplementation with vitamin E at moderate doses
protects against the teratogenic and toxic actions
of
vitamin A.
Supplementation with vitamin E increases liver
storage of vitamin A.
Vitamin E supplementation improves
plasma vitamin A levels in children
with vitamin A deficiency.
Vitamin E deficiency accelerates the

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 depletion of liver vitamin A stores.

VITAMIN B6
protein The level of protein intake is inversel correlated
with plasma B6 and pyridoxal phosphate levels,
and with the
urinary excretion of 4-pindoxic acid.

Diets providing the same amino acidpattern as

corn cause a decrease of B6concentration in plasma
and of PLP
in liver.
dietary fibre Ingestion of 15 g fibre per day during 18
days causes a fall in plasma levels of
B6 and PLP(pyridoxal phosphate) as well as a rise
in faecal
excretion of the vitamin.

VITAMIN E
vitamin C Vitamin C acts synergistically in the
intracellular antioxidant system to
regenerate reduced tocopherol.

IRON
protein Addition of protein to the diet (fish,poultry, meat)
increases the absorption of non-haem iron.
amino acids Different amino acid mixtures promoteiron
absorption. Cysteine is one ofthe most efficient.
organic acids Diets with relatively low pH or with elevated lactic
acid content enhance iron absorption.

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phosphate Calcium phosphate decreases ironabsorption, but
inorganic Phosphate has no effect.

zinc Use of Zn supplements inhibits iron absorption.

vitamin C Favours the absorption of non-haem iron by

binding and solubilizing it atthe physiological
intestinal pH.
Facilitates iron mobilization by inhibiting ferritin
breakdown at the lysosome.
Vitamin C deficiency causes iron accumulation as
haemosidenn.

vitamin A Vitamin A deficiency inhibits iron utilization and

accelerates the development of anaemia.
Iron deficiency is epidemiologicallyassociated
with vitamin A deficiency.
Vitamin A fortification improves the
haematological indices of populations.
polyphenols They bind and insolubilize iron. Vegetables
with high polyphenol content
may have low iron bioavailability.

ZINC
protein Favours Zn absorption by decreasing the inhibitory
action of phytates.
Malnourished children treated with soy-based
protein diets exhibit lower plasma Zn levels and
slower rate of
weight gain.

amino acids Several amino acids increase Zn absorption,

possibly by facilitating the mineral's release from
the
Ca-phytate-Zn complex.
Histidine is one amino acid that inhibits Zn
absorption, by forming insolublecomplexes. This
action may be antagonized by adding other amino
acids or protein to the diet.
iron Non-haem iron administration decreases inorganic
zinc absorption.
Haem iron has no inhibitory effect on Zn

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 absorption.
Supplementation of healthy infants with 30 mg
iron per day during 3months had no effect on
serum zinc
levels.

CALCIUM
protein Dietary protein stimulates urinary Ca excretion.

A moderate increase in dietary protein intake, from

65 to 94 g per day during28 days does not affect
calcium balance in
healthy subjects. [
fat Decreases Ca absorption by forming insoluble
soaps.

sodium Increases in NaCI intake increase urinary Ca

excretion and lower serum Ca in subjects with
hypercalciuria.
Low-sodium diets reduce urinary Ca excretion in
hypercalciuria individuals.

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