High fructose syrup :

High fructose corn syrup is a sweetener that is commonly used in the food and
beverage industry. It is produced by processing corn starch to convert some of its
glucose into fructose. The resulting syrup is sweeter than regular corn syrup, which is
primarily glucose. HFCS is used as a sweetener in a variety of processed foods and
beverages, including soft drinks, baked goods and more.
High fructose corn syrup (HFCS) is produced through a multi-step process that
involves the enzymatic conversion of corn starch into sugars. Here's a simplified
overview of the production process:
Corn Starch Extraction:
 Corn kernels are first cleaned and steeped in a water solution to separate the
starch from the other components.
 The extracted corn starch is then separated, washed, and dried
Liquefaction: The separated corn starch is then liquefied by adding enzymes. This
breaks down the starch into shorter chains of glucose.
Saccharification: Additional enzymes are added to further break down the starch
chains into individual glucose molecules.
Isomerization: In this step, the glucose-rich syrup undergoes a process called
isomerization. This involves the conversion of some of the glucose into fructose using
an enzyme called glucose isomerase. This step increases the fructose content in the
syrup.
Separation and Filtration: The syrup is then filtered to remove any impurities and
separate the desired components.
Purification: The syrup undergoes a purification process to remove any remaining
impurities and colour .
Blending and Refining:
The final step involves blending the glucose and fructose streams to achieve the
desired ratio. The most common forms of HFCS are HFCS-42 (42% fructose) and
HFCS-55 (55% fructose).
The syrup may undergo additional refining steps to remove impurities and achieve the
desired quality standards.
Concentration:
The syrup is then concentrated to the desired level by removing excess water,
resulting in a thick, sweet syrup.
Fructose is a monosaccharide widely used for food, pharmaceutical, and medical
applications.
High fructose syrup (HFS) can be synthesized from starch or inulin using
enzymatic/chemical methods.
(Chemical hydrolysis of starch/inulin can also be used for the production of HFS.
However, chemical approach is generally not used due to some drawbacks like
formation of Di fructose anhydrides and other coloured products which lower fructose
yield and demand an improved downstream processing and energy requirement.)
A well-known conventional approach is the hydrolysis of starch using amylolytic
enzymes and subsequent isomerization of dextrose to fructose by glucose isomerase.
The product yield by this method is only 42% and the product mixture also contains
50% dextrose and 8% other saccharides. HFS is prepared from starch by a multi
enzymatic method using α-amylase, glucoamylase, and glucose isomerase.
HFS can also be produced from inulin by a single-step method using inulinases. In the
single-step enzymatic method, inulinase acts sequentially on β-(2, 1) linkages of inulin
to release the fructose units. By this method approximately 95% fructose yield can be
obtained.
( inulin : Inulin is a type of dietary fiber that belongs to a class of compounds known
as fructans. It is found in a variety of plants, including chicory roots, Jerusalem
artichokes, dandelion roots, and agave. Inulin is a polysaccharide, meaning it consists
of multiple sugar molecules linked together .inulin is composed of fructose molecules
linked together by β(2-1) glycosidic bonds.)

Conventional Multi enzymatic Method for the Preparation of HFS from Starch
Conventionally, high fructose syrup is produced by multi enzymatic hydrolysis of corn
starch, involving the action of three enzymes, namely α-amylase, glucoamylase, and
glucose isomerase. The conventional production of HFS basically involves the
following four steps:
(a) liquefaction,
(b) saccharification of polymer starch into its monomeric units (dextrose),
(c) isomerization of dextrose to fructose,
(d) fractionation to enrich the final concentration of fructose by ion-exchange
method. Besides these steps, crystallization of fructose is also carried out to obtain dry
and crystalline fructose.
A . Starch liquefaction :
Liquefaction and partial hydrolysis of starch is the first step in the production of HFS
by the conventional method. In this step, starch obtained after wet milling of corn is
hydrolysed by thermostable α- or β-amylase (EC 3.2.1.1) into oligomers of
intermediate molecular weight.
Generally, α- or β-amylases are derived from thermostable bacterial strains like
Bacillus stearothermophilus, B. licheniformis, and B. subtilis.
B. stearothermophilus and B. licheniformis amylases are very thermostable and
usually used at temperature above 100 °C, while B. subtilis enzyme is a little less
thermostable and not used above 90 °C..
Alpha-amylase randomly disrupts the backbone of starch containing the α-1, 4
glycosidic linkages producing soluble, low- and medium-molecular weight
saccharides.
Enzymatic liquefaction requires specific regulating parameters such as starch solids,
pH, calcium level, reaction time, and temperature. Solid level is preferably maintained
at 30–35% to achieve complete starch gelatinization.
Starch slurry pH is normally controlled at 5.8–6.5. Higher pH increases byproduct
formation, while lower pH causes inactivation of enzyme which correspondingly
retards the product yield and increases refining cost. Calcium is used as a cofactor to
enhance amylase thermostability. It is added in chloride and oxide form at a
concentration of 100–200 ppm for B. stearothermophilus and B. licheniformis
amylases, but at 300 ppm for B. subtilis amylase. Reaction time and temperature are
generally balanced to provide optimal conditions for enzyme activity and starch
gelatinization.
Different processes like Enzyme-Heat-Enzyme (EHE), High Temperature (HT), and
Low Temperature (LT) are used to carry out enzymatic liquefaction.

B. Saccharification:
Saccharification is carried out to convert partially hydrolysed starch into dextrose
using glucoamylase (EC 3.2.1.3). Glucoamylase hydrolyses the di- and oligomeric
products formed by the action of amylase by cleaving the α-1, 4 and α-1, 6 glycosidic
linkages sequentially from the non-reducing end.
Glucoamylase for commercial purposes is traditionally derived from filamentous
fungi like Aspergillus, Rhizopus, etc. The type and number of linkages, size of the
molecule, and the arrangement of α-1, 4 and α-1, 6 linkages affect the rate of
hydrolysis by glucoamylase. Rate of hydrolysis increases with increase in the number
of linkages.
Approximately 95% of dextrose can be obtained after saccharification along with
other sugars like maltose (1%), isomaltose (1.5%), and maltulose (0.2%). Both
maltose and isomaltose are formed by reverse reaction catalysed by glucoamylase. For
commercial operation, minimizing this reversion is important in maximizing dextrose
yield. This can be attained by adding a high dose of glucoamylase and reducing the
reaction time so that the final hydrolysate can be processed rapidly to prevent
excessive reversion.
Dextrose yield can also be increased on combining glucoamylase with either
debranching enzyme from B. acidopullulyticus or the enzyme exhibiting both
amylase and transferase properties from B. megaterium . Saccharified hydrolysate is
then clarified by vacuum filtration and refined by combining carbon- and ion-
exchange technologies. The refined hydrolysate is processed to high dextrose corn
syrup, crystalline or liquid dextrose, or as feed for high fructose syrup production.
C. Isomerization :