BIOCHEMICAL OPERATIONS

CONTENTS

1. Fundamentals
2. Objective and application
3. Transformations of raw material
4. Technological factors
5. Equipments

Van Viet Man LE

Ho Chi Minh City University of
Technology 1

BIOCHEMICAL OPERATIONS
1. Fundamentals

 Chemical operation in which the reactions are

catalyzed by enzymes
 Enzyme source:
 Microbial enzymes (Inoculum/ contaminated
microbes): endoenzymes/ exoenzymes
 Enzyme in raw material: plant and animal
 Enzyme preparation

Van Viet Man LE

Ho Chi Minh City University of
Technology 2

1
 BIOCHEMICAL OPERATIONS
1. Fundamentals
 Concept: enzyme and enzyme preparation
 Catalytic activity of enzyme preparation
 Specific activity
 Immobilized enzyme
 Advantages

 Disadvantages

Van Viet Man LE

Ho Chi Minh City University of
Technology 3

BIOCHEMICAL OPERATIONS
2. Objective and application
 Application of enzyme preparation to food industry:
 Increase in catalytic activity of the enzymes in raw
material or of the microbial enzymes for shortening the
operation time or improving the reaction yield

 Replacement of chemical reactions by biochemical

reactions

Van Viet Man LE

Ho Chi Minh City University of
Technology 4

2
 BIOCHEMICAL OPERATIONS
2. Objective and application
 Preparation
Ex: Fruit juice processing
 Exploitation
Ex:
 Fruit juice processing

 Brewing, Wine-making

 Peptide production from whey

 Fish sauce/ soy sauce processing

Van Viet Man LE
Ho Chi Minh City University of
Technology 5

BIOCHEMICAL OPERATIONS
2. Objective and application
 Processing
Ex:
 Maltose, maltodextrin, glucose, high fructose syrup
production

 Invert sugar production

 Preservation
Ex: Fruit juice processing

 Improvement
Ex: Fruit juice processing, brewing, wine-making
Van Viet Man LE
Ho Chi Minh City University of
Technology 6

3
 BIOCHEMICAL OPERATIONS
3. Transformations of raw material
 Amylase and starch Amylose

A B
Van Viet Man LE
Ho Chi Minh City University of
Technology 7

C chain
(Reducing end) Amylopectin

Van Viet Man LE

Non-reducing
Ho Chi Minh City University of end
Technology 8

4
 Van Viet Man LE
Ho Chi Minh City University of
Technology 9

Crystal region in
starch granule

Van Viet Man LE

Ho Chi Minh City University of
Technology 10

5
 BIOCHEMICAL OPERATIONS
3. Transformations of raw material
 Amylase and starch
 Alpha amylase (EC 3.2.1.1): endo-enzyme

 Beta amylase (EC 3.2.1.2): exo-enzyme

 Glucoamylase (EC 3.2.1.3): exo-enzyme

 Pullulanase (EC 3.2.1.41)

 Isoamylase (EC 3.2.1.68)

 Alpha glucosidase (EC 3.2.1.20)

Van Viet Man LE
Ho Chi Minh City University of
Technology 11

BIOCHEMICAL OPERATIONS
3. Transformations of raw material
 Amylase and starch: selection of amylase preparation
Example:
 Glucose production
 Maltose production
 Maltodextrin production

Van Viet Man LE

Ho Chi Minh City University of
Technology 12

6
 BIOCHEMICAL OPERATIONS
3. Transformations of raw material
 Pectinase and pectin
Substrates:
 Protopectin
 Pectic acid
 Pectinic acid
 Pectin
 Pectate
 Pectinate

Van Viet Man LE

Ho Chi Minh City University of
Technology 13

BIOCHEMICAL OPERATIONS
3. Transformations of raw material
 Pectinase and pectin
Enzymes catalyzing pectin transformations
 Polymethylgalacturonase
 Endo-enzyme
 Exo-enzyme
 Polymethylgalacturonalyase (trans-eliminase)
 Endo-enzyme (EC.4.2.2.10)
 Exo-enzyme

Van Viet Man LE

Ho Chi Minh City University of
Technology 14

7
 BIOCHEMICAL OPERATIONS
3. Transformations of raw material
 Pectinase and pectin
Enzymes catalyzing pectic acid transformations
 Polygalacturonase
 Endo-enzyme (EC 3.2.1.15)
 Exo-enzyme 1 (EC 3.2.1.67)
 Exo-enzyme 2 (EC 3.2.1.82)
 Polygalacturonalyase (trans-eliminase)
 Endo-enzyme (EC.4.2.2.2)
 Exo-enzyme (4.2.2.9)
Van Viet Man LE
Ho Chi Minh City University of
Technology 15

BIOCHEMICAL OPERATIONS
3. Transformations of raw material
 Pectinase and pectin
Enzymes catalyzing oligo-D-galacturonate transformations:
 Oligogalacturonase
 Oligogalacturonalyase (trans-eliminase), (EC 4.2.2.6)
Pectin esterase (EC 3.1.1.11)

Van Viet Man LE

Ho Chi Minh City University of
Technology 16

8
 BIOCHEMICAL OPERATIONS
3. Transformations of raw material

 Pectinase and pectin: selection of pectinase preparation

Example:
Wine making

Van Viet Man LE

Ho Chi Minh City University of
Technology 17

BIOCHEMICAL OPERATIONS
3. Transformations of raw material
 Protease and protein
 Peptidase (EC 3.4.11-15)
 Aminopeptidase

 Carboxylpeptidase

 Dipeptidase

Van Viet Man LE

Ho Chi Minh City University of
Technology 18

9
 BIOCHEMICAL OPERATIONS
3. Transformations of raw material
 Protease and protein
 Proteinase (EC 3.4.21-24)
 Sero-protease: active site contains of his, ser,
asn (trypsin, chymotrypsin, elastase…)
 Sulfo-protease: active site contains –SH group
(papain, chymopapain, bromelain, ficin, some
microbial proteases)
 Acid protease : active site contains amino acid
with 2 carboxyl groups –COOH (pepsin, some
fungal proteases)
 Metallo-protease: active site contains metal
Van Viet Man LE
Ho Chi Minh City University of
Technology 19

BIOCHEMICAL OPERATIONS
3. Transformations of raw material
 Protease and protein: selection of protease

Example:
 Meat treatment
 Fish sauce processing

Van Viet Man LE

Ho Chi Minh City University of
Technology 20

10
 BIOCHEMICAL OPERATIONS
3. Transformations of raw material
 Gluco-isomerase

 Invertase

 Lactase

 Cellulase

 Hemicellulase

 Other enzyme preparation

Van Viet Man LE

Ho Chi Minh City University of
Technology 21

BIOCHEMICAL OPERATIONS
4. Technological factors
 Temperature

Van Viet Man LE

Ho Chi Minh City University of
Technology 22

11
 Van Viet Man LE
Ho Chi Minh City University of
Technology 23

BIOCHEMICAL OPERATIONS
4. Technological
factors
 pH

Van Viet Man LE

Ho Chi Minh City University of
Technology 24

12
 Van Viet Man LE
Ho Chi Minh City University of
Technology 25

BIOCHEMICAL OPERATIONS
4. Technological
factors

 Substrate

Van Viet Man LE

Ho Chi Minh City University of
Technology 26

13
 BIOCHEMICAL OPERATIONS
4. Technological factors
 Substrate concentration

V = Vmax*[S]/(Km+[S])

S: Substrate concentration
V: Reaction rate
Vmax: Maximum reaction rate
Km: Constant of Michaelis-Menten

Van Viet Man LE

Ho Chi Minh City University of
Technology 27

BIOCHEMICAL OPERATIONS
4. Technological factors
Michaelis-Menten equation

When V=Vmax/2 → Km=S

Van Viet Man LE
Ho Chi Minh City University of
Technology 28

14
 BIOCHEMICAL OPERATIONS
4. Technological factors

Disadvantages of Michaelis-Menten equation

Van Viet Man LE

Ho Chi Minh City University of
Technology 29

Determination of Km and Vmax: Lineweaver-Burk method

(1/V) = (1/Vmax) + (Km/Vmax)*(1/S)

Van Viet Man LE

Ho Chi Minh City University of
Technology 30

15
 BIOCHEMICAL OPERATIONS
4. Technological factors
 Activator

 Inhibitor:
 Competitive
 Non-competitive

Van Viet Man LE

Ho Chi Minh City University of
Technology 31

BIOCHEMICAL OPERATIONS
5. Equipments
 Bioreactor: vessel with mixer and sensors for adjustment
of temperature and pH (homogeneous systems)

 Evaluation of biochemical operation:

 Reaction yield
 Kinetics of S and P

Van Viet Man LE

Ho Chi Minh City University of
Technology 32

16