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Industrial Uses of Enzymes: Dr. Reda Mahgoub Mohamed

1. Enzymes play a key role in industrial starch conversion processes like glucose syrup production, high fructose corn syrup production, and ethanol production. Amylases like alpha-amylase and glucoamylase break starch down into sugars like glucose. Glucose isomerase converts glucose to fructose for high fructose corn syrup. 2. Different starch conversion processes produce varying sugar syrups. High maltose syrups contain 45-85% maltose while high conversion syrups contain 30-47% maltose and 35-43% glucose. 3. Ethanol is produced from starch via enzymatic liquefaction and saccharification of starch into

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159 views23 pages

Industrial Uses of Enzymes: Dr. Reda Mahgoub Mohamed

1. Enzymes play a key role in industrial starch conversion processes like glucose syrup production, high fructose corn syrup production, and ethanol production. Amylases like alpha-amylase and glucoamylase break starch down into sugars like glucose. Glucose isomerase converts glucose to fructose for high fructose corn syrup. 2. Different starch conversion processes produce varying sugar syrups. High maltose syrups contain 45-85% maltose while high conversion syrups contain 30-47% maltose and 35-43% glucose. 3. Ethanol is produced from starch via enzymatic liquefaction and saccharification of starch into

Uploaded by

Islam Imad
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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Industrial uses of enzymes

By

Dr. Reda Mahgoub Mohamed


1. Starch conversions
• Production of glucose syrup
• Production of high fructose corn syrup (HFCS)
• Production of high maltose conversion syrups
• Production of ethanol
1. Starch conversions
Starch contains about 15–30% amylose and 70–85% amylopectin.
Enzymes have largely replaced the use of strong acid and high temperature to break down
starchy materials.

Three types of enzymes are involved in starch bioconversion:

1.endo-amylase (α-amylase, EC 3.2.1.1), [Bacillus lichiniformis, Bacillus subtilis, and


Bacillus amyloliquefaciens and fungi such as Aspergillus oryzae]

2.exo-amylases
glucoamylase or glucan 1,4-a-glucosidase, EC 3.2.1.3 [Endomycopsis, Aspergillus,
Penicillium, Rhizopus, and Mucor]
β-amylase, EC 3.2.1.2) [Bacillus megaterium, Bacillus cereus, Bacillus polymyxa,
Thermoanaerobacter thermosulfurogenes, and Pseudomonas sp.]

3. debranching enzymes (pullulanase, EC 3.2.1.41) Aerobacter aerogenes


In amylose these are linked  -(1, 4)-, with the ring oxygen atoms all on the same side.
In amylopectin about one residue in every twenty or so is also linked -(1,6)- forming
branch-points.
 -1,4

hydrolyzes the a-1,4-glycosidic bonds


in starch from the non-reducing ends,
generating maltose. The enzyme is
unable to by pass the a-1,6 linkages and
leaves dextrins, known as b-limit Hydrolyse internal (endo) a- 1,4 but not
dextrins a- 1,6 producing malto-oligosaccharides

Enzymatic hydrolysis of amylose


cleaves glucose units from the non reducing end of starch
and it can hydrolyze both a-1,4 and a-1,6 linkages of
starch, slower

Enzymatic hydrolysis of amylopectin


Dextrins: A group of low-molecular-weight carbohydrates
produced by the hydrolysis of starch.

Dextrins are mixtures of linear a-(1,4)-linked D-glucose


polymers starting with an a-(1,6) bond.
Amylases break starch into sugars
All amylases are glycoside hydrolases and act on α-1,4-glycosidic bonds.

α-amylases
Both the salivary and pancreatic amylases are α-amylases. They are Ca metalloenzymes,
completely unable to function in the absence of calcium. They act at random locations along
the starch chain hence faster than

β-amylases
Working from the non-reducing end, β-amylase catalyzes the hydrolysis of the second α-1,4
glycosidic bond, cleaving off two glucose units (maltose) at a time. During the ripening of
fruit, β-amylase breaks starch into sugar, resulting in the sweet flavor of ripe fruit.

Both are present in seeds; β-amylase is present prior to germination, whereas α-amylase
and proteases appear once germination has begun.
Enzymes for starch conversion
α-amylase randomly hydrolyse α-1,4 linkages in both amylose and amylopectin to yield
mixture of glucose, maltose, maltotriose and series of α-limit dextrins.

β-amylase sometimes used in place of α-amylase. They hydrolyze alternate α-1,4 linkages and
yield maltose residues and β -limit dextrins.

Glucoamylase hydrolyses α-1,4 and α-1,6 linkages but is less efficient than α-amylase. Major
role is to break cross links of amylopectin resulting in complete breakdown to glucose.
Generally used to reduce CHO content of beers. Industrially obtained from fungus Aspergillus
niger.

Glucose isomerase is used for conversion of glucose obtained after processing to fructose.

Pullulanase (pullulan α-1,6-glucanohydrolase) cleaves the α-1,6- linked branch points of starch
and produces linear amylosaccharides of varying lengths.
1. Production of D-glucose from starch by acid hydrolysis (chemical) produces
undesirable bitter sugar (gentiobiose), and the inevitable formation of salt (from
subsequent neutralization with alkali) and coloring materials.

With the discovery and development of thermostable a-amylase from Bacillus


licheniformis, an enzymatic process has replaced the acid hydrolysis process.
liquefaction and saccharification
Typically, glucose syrups (DE 97–98) having 96% glucose contain 2–3%
disaccharides (maltose and isomaltose) and 1–2% higher saccharides.

Dextrose equivalent (DE) is a measure of the amount of reducing sugars present in a


sugar product, relative to glucose, expressed as a percentage on a dry substance basis
(DS). an estimate of the percentage reducing sugars present in the total starch
product
GLUCOSE ---- FURCTOSE ISOMERIZATION

2. HFCS
Glucose isomerase (also known as xylose isomerase, EC 5.3.1.5) is an example of the
highly successful application of enzyme biotechnology to an industrial process that has no
commercially viable route through conventional chemistry.

Chemical isomerization of glucose to fructose at high pH and high temperature leads to


undesirable side products, some of which are colored and have off flavors.

Enzyme- catalyzed isomerization (at moderate pH and temperature) does not form
undesired side products.

Streptomyces, Bacillus, Arthobacter, and Actinoplanes


3. Various maltose-containing syrups are used in the brewing, baking, soft drink,
canning, confectionery, and other food industries.

There are three types of maltose-containing syrups:

1.high-maltose syrup (DE 35–50, 45–60% maltose, 10–25% maltotriose, 0.5–3%


glucose).
2.extra high-maltose syrup (DE 45–60, 70–85% maltose, 8–21% maltotriose, 1.5–2%
glucose).
3.high conversion syrup (DE 60–70, 30–47% maltose, 35–43% glucose, 8–15%
maltotriose).
liquefaction and saccharification, as in the production of glucose.

However, in this process, the liquefaction reaction is terminated when the DE


reaches about 5–10 since a low DE value increases the potential for attaining high
maltose content.
maltogenic amylase such as β-amylase, β -amylase with pullulanase or isoamylase,
or a fungal α-amylase at pH 5.0–5.5 and 50–55o C.
4- Production of ethanol
The process of making ethanol from starch involves three basic
steps:

1. preparation of the glucose feedstock.


2. Fermentation of glucose to ethanol.
3. Recovery of ethanol.

Enzymes have major role in preparation of feedstock:


Corn kernels contain 60–70% starch
Wet milling process

corn is steeped in acidic water solutions (corn steep liquor) and the
oil, protein, and fiber fractions are successively removed as products
leaving the starch fraction.

Enzymatic liquefaction and saccharification of the starch fraction


are then carried out for the production of glucose

Glucose is fermented by the traditional yeast Saccharomyces


cerevisiae to ethanol, which can be recovered by distillation.
Dry grinding process
lower capital investment required in comparison to that of wet mills.

In the typical dry grind process, corn is mechanically milled to a coarse flour. Oil, protein,
and fiber fractions are not isolated

liquefaction, enzymatic saccharification using glucoamylase

fermentation using the conventional yeast are carried out simultaneously.

The addition of protein-splitting enzymes (proteases) releases soluble nitrogen compounds


from the fermentation mash and promotes growth of the yeast, decreasing fermentation time.

The residue left after fermenting the sugars is known as distiller’s grains, which is used as
animal feed.
Milled grain
steam

Gelatinized material
cool
a-amylase
Liquefied material Bacillus amyloliquefaciens
Glucoamylase

Saccharified material
Yeast
Glucose isomerase

Fructose Fermentation

Alcohol
Industrial production of alcohol and fructose from starch

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