Immobilization techniques, Immobilization techniques in food industry, Immobilized Enzymes, Need for immobilization, Role of immobilized Enzymes in Food Industry, Methods of immobilization, Production of lactose free milk, Production of High Fructose corn syrups, Production of Juice in industry level by Immobilized enzymes of Pectinase, Meat tenderization by immobilized Enzymes, Immobilized Amino acylase, immobilized glucose isomerase, immobilized pectinase, Immobilized alkaline phosphatase.

1. Role of immobilized / bound
Enzymes in Food industry
Jasmine Juliet .R
Teaching Assistant
Biotechnology Dept
AC&RI, Madurai.

3. Immobilization Techniques
 Imprisonment of cell or enzyme in a distinct support / matrix.
 The support / matrix allows exchange of medium.
 The medium contains substrate or effector or inhibitor molecules.
 First immobilization technology: Aminoacylase by Aspergillus
oryzae for the production of L-aminoacids in Japan.
 Two main advantages of enzyme immobilization:
1. Increased functional efficiency
2. Enhanced reproducibility

4. Immobilization Techniques in Industrial
Applications
• Immobilised enzymes have been fixed to a static surface in order to improve
the efficiency of the catalysed reaction.
• Immobilised enzymes are utilised in a wide variety of industrial practices:
• Biofuels – Enzymes are used to produce ethanol-based fuels.
• Medicine – Enzymes are used to identify diseases and pregnancy.
• Biotechnology – Enzymes are involved in gene splicing.
• Food production – Enzymes are used in the production of dairy products.
• Textiles – Enzymes are utilised in polishing cloth.

6. Immobilized Enzyme - Introduction
 Enzymes are macromolecular biocatalysts, widely used in food industry.
 In applications, enzymes are often immobilized on inert and insoluble
carriers, which increase their efficiency due to multiple reusability.
 The properties of immobilized enzymes depend on the immobilization
method and the carrier type.
 The choice of the carrier usually concerns the biocompatibility, chemical
and thermal stability, insolubility under reaction conditions, capability of
easy regeneration and reusability, as well as cost efficiency.

9. Need for immobilization
• Accelerates the chemical reaction.
• Specificity and un-modified Enzyme.
• Cost effective.
• Not difficult to separate.
• Attachment to polymers/matrix, causes re-use.

11. Applications of immobilized
Enzymes
• Immobilized enzyme-Aminoacylase used for the production of
L-aminoacids.
• In food industry, Fructose syrup is produce from glucose by use of
immobilized enzyme glucose isomerase.
• Accurate analysis of sample done with the help of specific
immobilized enzymes.
• Immobilized enzymes in industry for the production of various
industrial products.

12. Applications of immobilized
Enzymes
Immobilized Enzymes Industrial Products
Aminoacylase Aminoacid
α- Amylase, Glucoamylase Glucose from starch
Glucose isomerase Fructose from Glucose
Penicillin Amidase 6-Aminopenicillanic acid from
Penicillin
Β- Galactisidase Hydrolysis of lactose in milk or
whey

14. Role of immobilized Enzymes in
Food industry
 The enzyme catalyzed process technology has enormous potential
in the food sectors.
 Enzymes may be used in different food sectors like:
o Dairy,
o Fruits & vegetable processing,
o Meat tenderization, fish processing,
o Brewery and wine making,
o Starch processing and many other.

15. Applications of immobilized Enzymes in
food industry
• In Food industry,
 Starch Hydrolysis.
 Production of High Fructose corn syrup.
 Use of Proteases.
 Production of Aminoacids.
 Antibiotics production.

16. Role of immobilized Enzymes in Food
industry
• The production of high fructose corn syrups was
greatly facilitated by the use of immobilized
glucose isomerase.
• Similarly, in Japan, the fermentation industry
proved its processing efficiency for amino acids
through the use of immobilized amino acid acylase.

17. Role of immobilized Enzymes in Food
industry
• Pectinase breakdown substances in apple cell walls
and enable greater juice extraction.
• Lactase breakdown lactose in milk into glucose and
galactose .
• This makes milk drinkable for lactose intolerant
people.

18. (I) Methods of production of lactose-free
milk and its advantages
• Lactose is a disaccharide of glucose and galactose which can be
broken down by the enzyme lactase
• Historically, mammals exhibit a marked decrease in lactase
production after weaning, leading to lactose intolerance
• Incidence of lactose intolerance is particularly high in Asian,
African and Aboriginal populations
• Incidence is lower in European populations (due to a mutation that
maintains lactase production into adulthood)

20. Producing Lactose –Free milk
• Lactose-free milk can be produced by treating the milk with the
enzyme lactase.
• The lactase is purified from yeast or bacteria and then bound to an
inert substance (such as alginate beads)
• Milk is then repeatedly passed over this immobilised enzyme,
becoming lactose-free.
• This involves splicing the lactase, so that the lactose is broken
down prior to milking.

21. Breakdown of Lactose by the Enzyme Lactase

22. Generation of Lactose-Free Milk Using
Immobilised Enzymes

23. Milk, lactase enzyme and glucose test strips

24. Advantages of Lactose-Free Dairy
Products
• The generation of lactose-free milk can be used in a variety of ways:
 As a source of dairy for lactose-intolerant individuals
 As a means of increasing sweetness in the absence of artificial
sweeteners (monosaccharides are sweeter tasting)
 As a way of reducing the crystallisation of ice-creams (monosaccharides
are more soluble, less likely to crystalise)
 As a means of reducing production time for cheeses and yogurts
(bacteria ferment monosaccharides more readily)

25. (II) High-Fructose Corn Syrup
• The most important use of immobilized enzyme in food
industry is the conversion of glucose syrups to high-
fructose corn syrup (HFCS) by the enzyme glucose
isomerase.
• The enzyme is immobilized and proceeds to the
conversion of glucose into fructose makings it sweeter.
• In many countries, where sugar prices are high, HFCS is
used as a sweetening agent

26. Immobilized Glucose Isomerase
enzyme
• Glucose isomerase (G.I., E.C. 5.3.1.5), also known as xylose-isomerase,
D-xylose isomerase, & D-xylose keto-isomerase, is an isomerase that
isomerizes aldose, such as D-xylose, D-glucose, D-ribose to the
corresponding ketone.
• The chemical name of this enzyme class is D-xylose aldose-ketose-
isomerase enzyme.
• Glucose isomerase has a wide range of sources, including
microorganisms, such as bacteria and fungi, as well as plants and animals.
• Isomerase converts D-glucose to D-fructose.

27. High Fructose Corn Syrups Applications
• Isomerase is used in the production of high fructose
syrups from glucose syrups (which are them salves
usually derived from maize or corn starch).
• The glucose isomerase is used on an extensive scale
throughout the world in the production of high fructose
syrups for the confectionery and soft drinks industries.

29. (III) Industrial Production of Fruit
Juices
• Cell wall of the fruits contains pectins and
carbohydrates which hold plant together.
• Immobilized pectinase, hydrolyse pectin and thus
increases the amount of fruit juices with no
residues of pectin.

30. Immobilization of Pectinases by Sequential
Layering on Chitosan Beads
• Chitosan beads were prepared as a support material for immobilization of
pectinases using a sequential layering approach.
• Three layers of pectinases were covalently immobilized on the support.
• The results suggested that increasing the density of pectinases on chitosan
support might have decreased the catalytic ability of enzymes due to either
restriction to the protein backbone or due to substrate accessibility
limitations.
• Therefore, optimal protein loading should be pursued for utilization of
immobilized pectinases with highest specific activity.

32. (IV) Meat tenterization
o Proteases have been used in food processing.
o Protease enzymes play a prominent role in meat tenderization, especially of
beef.
o An alkaline elastase and thermophilic alkaline protease have proved to be
successful and promising meat tenderizing enzymes, since they can hydrolyze
connective tissue proteins as well as muscle fiber proteins.
o A patented method used a specific combination of neutral and alkaline
proteases for hydrolyzing raw meat, possibly because the preferential
specificity was favorable when metalloprotease and serine protease were used
simultaneously.

33. Meat tenterization by immobilized
Alkaline Protease
• Alginate was found to be the best matrix for cell entrapment and
alkaline protease production, showing the highest specific productivity
and enzyme production followed by cells immobilized in agar and gelatin.
• Furthermore, the production of alkaline protease by Bacillus sp.
immobilized in alginate gel was enhanced by influence of various
parameters on alginate beads preparation including alginate concentration,
bead size, and biomass loading.
• Maximum enzyme production and specific productivity were achieved
using alginate.

35. Role of immobilized / bound Enzymes in
Food industry- Conclusions
• In recent years, significant progress in design of enzyme
immobilization, are developed.
• Designing ideal support material by modifying specific
structural features required for a target enzyme is now possible
by new simulations.
• It is our view that the future holds significant promise with
increased usage of immobilized enzymes in food, and other
industrial fields.

36. Thank you