Jordan Journal of Agricultural Sciences, Volume 8, No.

3 2012

Improvement of Yogurt Properties by Microbial Transglutaminase

Waleed Ahmed Mahmood* and Nawal Hurmiz Sebo**

ABSTRACT
Yogurt was manufactured from cow's milk which was treated with different concentrations of free and
immobilized Streptoverticillium mobaraense transglutaminase. The highest pH and lowest titratable acidity
during three weeks of storage were detected in the sample treated with 75 U/l of free transglutaminase before
starter addition. The concomitant MTGase treatment with starter addition resulted in significant increase in
yogurt gel strength and viscosity and decreased water separation. This effect was continued during storage at 4
C. The effect of enzymatic treatment with 45 U/l almost resembled that of adding 3% of skimmed milk powder.
Immobilized MTGase treatment had less effects on yogurt properties. Sensory evaluation of yogurt showed that
treated samples were superior to untreated sample in all sensory aspects through storage periods. To reveal the
occurrence of cross-linking among protein molecules, SDS-PAG electrophoresis was performed for yogurt
proteins. New high molecular weight protein bands had occurred accompanied by decreasing in casein and
whey proteins bands density.
Keywords: Transglutaminase, Yogurt, Streptoverticillium mobaraense.

soft cheeses and yogurt wherein syneresis is reduced.

INTRODUCTION

Among the conventional methods of improving yogurt

Texture and reological properties of some dairy

texture, is the addition of non fat dry milk (NFDM) and

products, such as soft cheeses, ice cream and yogurt, are

stabilizers to the milk (Guzman-Gonzalez et al., 1999;

affected mainly by the balance among the milk

Shah, 2003), but since the addition of stabilizers is not

components. Increasing protein level leads to an increase

permitted in many countries, alternative methods to

in gel strength. Yogurt and cheese usually suffer water

achieve texture improvement have been of interest in

separation during storage due to syneresis. The separated

recent years.

water

Transglutaminase (TGase) is an enzyme that

contains substantial amounts of proteins.

water

catalyzes the cross linking of proteins through the

separation objectionable (Han et al., 2003). Therefore,

formation of covalent bonds using two amino acids,

there remains a need for improved methods to produce

glutamine and lysine, enabling the proteins to behave

Moreover,

consumers

generally

find

such

like larger molecules. The cross-links introduced by this

* Ass. Prof., Dept. of Food Sci., College of Agric. and
Forestry, Mosul Univ., Iraq. Tel: +9647701667076
E-mail: waleedahmed53@yahoo.com
** Dept. of Food Tech., College of Agric. Salahaddin Univ.,
Iraq.Tel: +9647504451952
E-mail: nawal_1952@yahoo.com
Received on 15/6/2011 and Accepted for Publication on
5/3/2012.

enzyme change the protein structure and improve its

functional properties, like texture, without decomposing
the nutritional quality of the lysine residue (Seguro et
al., 1996). Since microbial TGase (MTGase) is available
in bulk from fermentation processes, the enzyme has

-333-

2012 DAR Publishers/University of Jordan. All Rights Reserved.

Improvement of Yogurt

Waleed Ahmed Mahmood and Nawal Hurmiz Sebo

been approved by food industries to improve quality of

absorbance was recorded at 525 nm using CECIL 3021

many foods such as meat, fish, soy products, yogurt,

spectrophotometer. One unit of transglutaminase activity

cheese, and ice cream (Zhu et al., 1995; Motoki and

was defined as the amount of enzyme which causes the

Seguro, 1998; Kuraishi et al., 2001; Jaros et al., 2007).

formation of one micromole of hydroxamic acid per

The

present

paper

describes

the

effect

of

minute at 37C. A calibration curve was prepared using

incorporation of the free and immobilized MTGase on

L-glutamic -monohydroxamate.

the properties of yogurt as a function of the added

Immobilization of transglutaminase: The enzyme

enzyme concentration and storage period .

was entrapped in calcium alginate gel beads following

the procedure of Fraser and Bickerstaff (1997). The

MATERIALS AND METHODS

enzyme solution was mixed with a same volume of 2%

Bacterial cultures and growth conditions: A

sodium alginate solution. The mixture was transferred to

lyophilized culture of Streptoverticillium mobaraense

a 10-ml syringe and drop wise into a beaker containing

(DSM-40847) was obtained from DSMZ, Germany. The

0.15 M calcium chloride solution with gentle stirring.

bacteria were propagated through submerged cultivation

Gel beads of calcium alginate were formed which left

( 30 C, 100 rpm) in a medium contained 2% starch, 2%

for 60 minutes for hardening, then they were collected,

peptone, 0.2% yeast extract, 0.2% dipotassium hydrogen

washed with distilled water, suspended in 0.1M, pH 6

phosphate, 0.1% magnesium sulfate and 0.05% glycerol

Tris-acetate buffer and stored at 4 C. To determine the

(Ando et al., 1989). Yogurt starter culture consisted of

immobilization efficiency, the beads gel was dissolved

and

by immersion in 0.05 M EDTA, pH 7 for 60 minutes and

Lactobacillus delbueckii subsp. bulgaricus was from

the activity of the obtained free enzyme was determined.

Streptococcus

salivarius

subsp.

thermophilus

Rhodia Food.

Milk treatment and yogurt manufacturing: Whole

Enzyme isolation and precipitation: At the end of

fresh cow's milk (containing 3.7% fat, 3.38% protein and

incubation period (3 days), the Streptoverticillium

12.44% total solids) was heated to 90 C for 5 minutes,

mobaraense culture was filtered off. The enzyme was

cooled down to 43 C and divided into 4 samples. To the

precipitated from the supernatant by adding two volumes

first sample, the free MTGase was added at the same

of cold acetone. The precipitate was dissolved in Tris-

time of the starter addition. The second sample was

acetate buffer (0.1M, pH 6) and lyophilized.

treated with MTGase as above and incubated at 40 C

Determination of transglutaminase activity: The

for 60 minutes then the enzyme was inactivated by

activity was measured by the colorimetric hydroxamate

heating to 90 C for 5 minutes, then the yogurt starter

procedure with N-carbo-benzoxy-L-glycine as the

was added at 43 C. To the third sample, the

substrate according to the method of Folk (1970).

immobilized enzyme beads were added, incubated at 40

Reaction mixture containing 50 l of the enzyme, 350 l

C for one hour, then the beads were removed by

of Tris-acetate buffer (0.1 M, pH 6), 25 l of 2 M

filtration through cheesecloth and the starter was added

hydroxylamine and 75 l of 0.1 M CBZ-L-GLN-GLY,

at 43 C. To the last sample, non-fat dry milk (NFDM)

was incubated at 37 C for 10 minutes and then the

was added without MTGase addition. These samples

reaction was stopped by adding 500 l of 15%

were used to manufacture yogurt according to the

trichloroacetic acid (TCA) containing 5% FeCl3. The

method of Tamime and Robinson (1999). Yogurt

-334-

Jordan Journal of Agricultural Sciences, Volume 8, No.3 2012

SAS (2001).

samples were then cooled and kept at 4 C to the next

day for analysis. Concerning the effect of the MTGase

RESULTS AND DISCUSSION

treatment

the

The effects of milk treatment with transglutaminase

concentration of the added enzyme in all cases was 75

or non-fat dry milk (NFDM) on the properties of the

unit/l. The set yogurt samples were stored at 4 C for 21

produced yogurt are summarized in Table (1).

on

the

stored

yogurt

properties,

Most MTGase treatments, with the different methods

days and the samples were weekly analyzed for acidity,

of application, caused non-significant increase (p0.05)

pH, physical and sensory properties.

Chemical analysis of milk and yogurt: pH of

in yogurt pH values as compared to the untreated yogurt

yogurt was measured by pH-meter (WTW-530).

and with the yogurt containing NFDM. The effect of

Titratable acidity of milk and yogurt was estimated

enzymatic treatment on pH and acidity was found to be

according to IDF/ISO Standard (1991) by calculating

proportional to the concentration of the added enzyme.

SH value which is defined as the volume (ml) of 0.25

The highest pH (4.53) was detected in the sample treated

M NaOH that neutralize the acidity in 100 ml of milk.

with 75 U/l of free transglutaminase before the addition

To detect the formation of cross-links, SDS-PAG

of the starter in which the increase in pH became

electrophoresis was conducted for yogurt proteins by

significant upon storage at 4 C for three weeks and it

using vertical electrophoresis apparatus (Jookoh Co.

was accompanied by significant decrease in titratable

LTD) following the procedure described by Laemmli

acidity. A significant drop in all samples pH was

(1970).

observed upon increasing the storage period (Table 2).

Physical tests of yogurt: Yogurt gel firmness was

The expected explanation for this relationship is that

measured using texture analyzer (Stevens-LFRA with

the amino acids and small peptides, which are needed for

TA26 probe). To measure the quantity of separated

the growth of lactic acid bacteria, are cross linked by the

water, the procedure described by Harwalker and Kalab

transglutaminase treatment and became unavailable to

(1983) was followed by centrifuging 25 gm of yogurt

the bacteria. This might led to the retardation in bacterial

sample at 1400 g and 5 C for 15 minutes. Yogurt

growth and decrease in acid production. The same

viscosity was estimated using Bostwick consistometer

observation was found by Lorenzen et al. (2002) who

by measuring the distance which the stirred yogurt

stated that the pre incubation of milk with MTGase

sample moves on a declined metallic surface in 10

followed by thermal inactivation of the enzyme caused

seconds.

the

elongation

of

fermentation

time

while

the

Sensory evaluation: Sensory evaluation of the

concomitantly addition of the enzyme with the starter

produced yogurt was carried out for appearance, texture,

did not affect the acidity of the yogurt. On the other

flavor and taste after 1, 7, 14 and 21 days of storage

hand, Bonisch et al. (2007) detected that 88% of cross

according to Nelson and Trout (1964).

linking had taken place in the pH range of 5.7-6.6 and

the enzyme was inhibited at pH lower than 4.5.

Statistical analysis: The data were analyzed by

using the Complete Random Design (CRD) according to

-335-

Improvement of Yogurt

Waleed Ahmed Mahmood and Nawal Hurmiz Sebo

Table (1): Effect of treatment with transglutaminase or non-fat dry milk (NFDM) on some properties of yogurt.
Treatment

MTGase

pH

Acidity (SH)

strength

Viscosity

Syneresis

(gm/cm )

(cm/10 sec)

(ml/25gm)

conc.(U/l)
Control

Gel

4.36a

39.1cd

8.66n

10.41a

10.00a

Addition

of

free

15

4.37a

38.75d-f

9.66mn

9.75b

8.33c

MTGase

with

the

30

4.40a

38.51e-g

13.66gh

7.83gh

7.20ef

45

4.42a

38.40e-h

15.33ef

6.25k

6.50gh

60

4.43a

38.26f-h

21.00c

5.23l

6.40hi

75

4.45a

38.10g-i

29.00a

4.50m

6.10i

free

15

4.42a

38.53e-g

9.33mn

9.83b

9.10b

MTGase before the

30

4.47a

38.16g-i

11.33k

8.63e

7.63d

starter

45

4.49a

38.03h-j

14.33fg

8.33ef

7.06f

60

4.51a

37.73ij

16.00e

7.45hi

6.73g

75

4.53a

37.60j

22.33b

7.10ij

6.10i

of

15

4.40a

38.78de

9.33mn

10.01ab

9.80a

immobilized MTGase

30

4.43a

38.61e-g

11.66i-k

9.16c

9.10b

before the starter

45

4.44a

38.49f-h

14.00g

8.75de

8.56c

60

4.45a

38.35e-h

14.66fg

8.10fg

8.30c

75

4.48a

38.21e-h

17.33d

7.18ij

7.60d

4.35a

39.26gh

10.00lm

9.63b

9.76a

4.33a

39.56bc

11.00kl

9.13cd

9.16b

4.31ab

39.75ab

12.66hi

8.5ef

8.36c

4.30ab

39.80bc

12.33ij

7.83gh

7.70d

4.29b

40.00a

12.66hi

7.00j

7.40de

starter

Addition

of

Addition

Addition of NFDM
(%)

1.5
2
2.5
3

Means within columns followed by different letters are significantly different at p 0.05.
Concerning the effects of MTGase treatment on

enzyme-substrate affinity especially with high molecular

physical properties of the product, a significant increase

weight substrates where the mass transfer was reduced

(p0.05) in yogurt gel strength was detected upon the

leading to the obstruction of the substrate to reach the

treatment with all methods except with low enzyme

active site of the enzyme (Schorch et al., 2000). The effect

concentration (15 U/l). This increase was proportional to

of adding 30 U/l of free enzyme almost resembled the

the

enzyme. The

effect of adding 3% of NFDM (Table 1). Lorenzen et al.

directaddition of 75 U/l of free MTGase along with the

(2002) found that the treatment of milk with MTGase had

starter was significantly superior to all other treatments.

reduced water separation and improved the texture of set

The least effect was detected with immobilized enzyme

style yogurt. During the first week of storage, all free

treatment which could be due to the decrease in the

MTGase-treated samples (75 U/l) showed significant

concentration of

the

added

-336-

Jordan Journal of Agricultural Sciences, Volume 8, No.3 2012

increase (p0.05) in gel strength but the highest strength

results in a finer and stronger gel network in the yogurt

was detected in the sample to which free enzyme was

as compared with acid-induced gels. This indicates that

added concomitantly with the starter in which strength

the concomitantly addition of MTGase with the starter

increase was continued during all storage period.

stimulated the crosslinking of protein molecules which

The addition of immobilized enzyme caused non-

was continued during and after fermentation resulting in

significant increase during storage (Table 2). Schorch et

the formation of new covalent bonds which caused

al. (2000) demonstrated that after a certain time period,

the development of gel stiffness during storage period

the firmness of MTGase-induced gels reached nearly a

(Ozer et al., 2007). The enzymatic crosslinking of

plateau value, suggesting that most of the protein

proteins by TGase is considered as an important way for

connections were already built up during this time

improving the texture of yogurt and other protein-

period. They also

containing foods (Tamime, 2006; Jaros et al., 2007).

stated

that the basic function of

TGase is to cross-link milk proteins covalently, which

Table (2): Effect of transglutaminase treatment (75 U/l) on some properties of stored yogurt stored at 4C for 21 days.
Treatment

Storage

pH

Acidity (SH)

period (day)
Control

Gel

strength

Viscosity

Syneresis

(gm/cm )

(cm/10sec.)

(ml/25gm)

4.39de

39.10k

8.66k

9.75a

9.10a

4.25g

45.30f

10.66jk

8.75b

8.30b

14

4.17h-j

48.03c

10.66jk

8.45b

8.16b

21

3.91l

49.16a

10.66jk

8.35b

8.06b

Addition of free

4.49ab

38.00m

24.33c

4.50f

5.76e-g

MTGase with the

4.35ef

43.30i

38.33b

3.81g

5.23fg

starter

14

4.24g

45.52f

42.00a

3.61g

5.13g

21

4.15ij

47.70d

42.33a

3.46g

5.03g

Addition of free

4.51a

37.60n

14.33hi

6.61c-e

7.03cd

MTGase

4.40cd

42.65j

17.66ef

6.73c-e

6.36de

14

4.25g

45.33f

17.66ef

6.80c-e

6.30de

21

4.18hi

46.80e

17.00f

6.80c-e

6.16e

4.46bc

38.65l

12.62ij

6.88c-e

7.20c

immobilized

4.34ef

43.80g

13.66hi

6.90cd

6.80cd

MTGase

14

4.22gh

46.92e

14.00hi

6.96c

6.70cd

21

4.12i

48.26c

13.66hi

6.63c-e

6.56cd

before

the starter
Addition

of

Means within columns followed by different letters are significantly different at p 0.05.
Cross-linked yogurt with all treatments showed a

MTGase concentrations as compared to untreated

highly viscous structure especially at high added

sample and that contained non-fat dry milk. The highest

-337-

Improvement of Yogurt

Waleed Ahmed Mahmood and Nawal Hurmiz Sebo

had occurred upon

to the decreased permeability which had occurred as a

concomitantly addition of 75 U/l of free MTGase with

result of the formation of permanent cross-links among

the starter. All treated samples showed significant

the milk proteins. The decreased permeability causes

increase (p0.05) in viscosity except the treatment with

more compact and stable microstructure with smaller

low concentration of immobilized enzyme (Table 1).

compartments in yogurt which allowed more free water

The viscosity had continued to increase during the first

to be entrapped in the yogurt gel network (Moon and

week of storage after which lower increase was noticed

Hong, 2003). Motoki and Seguro (1998) mentioned that

during the remaining storage period with some

syneresis could be reduced by MTGase treatment which

variations among the treatments (Table 2). This could be

improves the water holding capacity of yogurt gel

attributed to differences in samples pH which may alter

network. On the other hand, the slow development of

the protein-protein interactions resulting in slow protein

acidity had an important role in decreasing water

rearrengments in the acid casein gels during cold storage

separation (Lorenzen et al., 2002).

viscosity (4.5 cm/10 sec.)

The

(Ozer et al., 1998). Kuraishi et al. (2001) observed

addition

of

the

free

MTGase

in

all

increased yogurt viscosity upon increasing the added

concentrations along with the starter was found to be the

enzyme concentration. They mentioned to the benefit of

most effective way for improving all physical properties.

this treatment in decreasing non fat solids content and

Treatment with the immobilized enzyme caused less

reducing the production cost.

effect than the free form. This may be due to the

Syneresis, or whey separation, is an important factor

physical obstruction caused by the alginate gel lattice

that affects consumer acceptance of yogurt. In unstored

against the movement of substrates and products

yogurt, MTGase treatment with all methods and

throughout the gel resulting in retarding and lowering

concentrations caused significant decrease (p0.05) in

the enzyme reaction rate. On the other hand, the

water separation except the treatment with 15 U/l of

immobilized enzyme treatment is unable to be applied

immobilized enzyme. The least quantity of separated

concomitantly with the starter and it is only applicable

water (6.1ml/25 gm) was detected upon treating with 75

by adding it prior to the addition of the starter.

U/l of free MTGase along with the starter as compared

The effect of MTGase treatments and storage periods

to the untreated sample (10 ml/25 gm). The addition of

on sensory evaluation of yogurt is indicated in Table (3).

NFDM had also reduced water separation. The effect of

All MTGase-treated samples during all storage periods

adding 3% of NFDM (7.4 ml/25 gm) resembled the

were superior to the untreated yogurt in all sensory

effect of the treatment with 45 U/l of free MTGase

properties. The treated samples were characterized by

(Table 1). The treatment with 75 U/l of free MTGase

dry and soft surfaces which could be attributed to the

along with the starter resulted in minimal separated

formation of glutamyl-lysine bonds and their effect in

water (5.03 ml/25 gm) at the end of storage period. On

improving the water-holding capacity of the yogurt gel

the contrary, the untreated sample had lost 9.1 ml/25 gm.

(Motoki and Seguro, 1998). A similar observation was

at the end of the storage period (Table 2). This was due

reported by Kuraishi et al. (2001).

-338-

Jordan Journal of Agricultural Sciences, Volume 8, No.3 2012

Table (3): Effect of transglutaminase treatment (75 U/l) on some sensory properties of stored yogurt stored at 4C
for 21 days.
Treatment

Storage period

Appearance

Texture

Flavor

Sour

(day)

(10)

(30)

(45)

(15)

(100)

8.00cd

20.00f-h

38.75ab

13.00ab

79.75d-g

8.25b-d

23.75c-g

41.75ab

14.00a

87.75a-d

14

7.75cd

19.50i

38.50ab

10.50e

76.25e-g

21

7.00d

19.25i

36.00b

10.50e

72.75g

Addition of free MTGase with

9.50ab

26.75a-c

35.75b

10.75de

82.75b-f

the starter

10.00a

28.00ab

40.00ab

11.75b-e

89.75a-c

14

9.75ab

28.75a

41.50ab

12.75a-c

92.75a

21

9.25a-c

26.75a-d

38.50ab

13.25ab

87.75a-d

9.50ab

25.50a-e

36.00b

10.75de

81.75c-f

10.00a

28.25a

40.25ab

12.00b-e

90.50a-c

14

9.75ab

27.75a-c

41.75ab

12.75a-c

92.00ab

21

9.25a-c

26.25a-d

38.25ab

13.25ab

87.00a-d

9.25a-c

23.25d-h

37.25ab

11.25c-e

81.00c-g

10.00a

26.00a-d

40.00ab

12.25b-d

88.25a-d

14

9.25a-c

25.75a-e

40.75ab

12.50a-c

88.25a-d

21

9.25a-c

25.00a-e

37.25ab

12.50a-c

84.00a-e

Control

Addition

of

free

MTGase

before the starter

Addition

of

immobilized

MTGase

taste

Total

Means within columns followed by different letters are significantly different at p 0.05.
analysis of this property and was in agreement with

Seven days were found to be a preferable period of

all treatments to get the best

Kuraishi et al. (2001) who stated that the sensory

appearance of the product. There was non-significant

evaluated texture of MTGase-treated yogurt was highly

decrease in this property in all treatments upon

superior to the untreated yogurt.

yogurt

storage

for

increasing the storage time. The least degree was

During the first week of storage, the flavor of the

recorded for the untreated sample after storage for 21

untreated sample was found to be superior to the

days.

MTGase-treated samples. This could be due to the effect

Concerning the sensory evaluation of yogurt texture,

of the enzymatic treatment on the obstruction of the

it was found that the MTGase-treated samples were

growth of lactic acid bacteria and retarding the

superior to the untreated sample throughout all storage

development of acidity and the production of flavor

periods. The best texture was obtained by the

compounds such as acetaldehyde, acetoin and diacetyl

concomitantly addition of free MTGase with the starter

through the first days of storage (Ozer et al., 2007).

but the differences among the treatments were non-

After two weeks of storage, the treated samples retained

significant. This result was in accordance to instrumental

the preference in the flavor property. Treated yogurt

-339-

Improvement of Yogurt

Waleed Ahmed Mahmood and Nawal Hurmiz Sebo

samples were also characterized by a fatty flavor. This

intensity of high molecular weight polymers bands

was in agreement with Metwally (2007) who observed

depending on the degree of cross-linking that occurred

that the treatment with MTGase produces fatty flavor

according to the treatment type. The highest cross-

which can replace some of added fat to the low-fat dairy

linking was observed upon the addition of 75 U/l of free

products.

enzyme along with the starter followed by the addition

The effect of MTGase treatment on yogurt protein

of the same concentration prior to the starter addition. A

molecules of one day-old yogurt was revealed by SDS-

relatively lower cross-linking had occurred upon using

PAG electrophoresis (Fig. 1). The treated yogurt was

the immobilized MTGase which could be due to the

characterized by a decrease in the intensity of casein and

effect of the alginate gel lattice on lowering the

whey protein bands accompanied by increasing the

enzymatic reaction rate.

Fig. 1: SDS-PAGE profile of MTGase-treated yogurt proteins. 1-Control,

2- Addition of 30 U/l of free

MTGase with the starter, 3- Addition of 75 U/l of free MTGase with the starter, 4- Addition of 75 U/l of free
MTGase before the starter, 5- Addition of 75 U/l of immobilized MTGase before the starter.

REFERENCES
Ando, H.; M. Adachi; K. Umeda; A. Matsuura; M. Nonaka; R.

cross-linking

Uchio; H. Tanaka and M. Motoki, 1989, Purification and

during

microbial

fermentation.

Food

Hydrocolloids, V. 21, pp. 585-595.

characterisation of a novel transglutaminase derived from

Folk, J. K., 1970, Tranglutaminase, in: Methods in

microorganisms. Agric. Biol. Chem., V. 53, No. 10, pp.

Enzymology, edited by Tabor, H. and C.W. Tabor, V. 17.

2613-2617.

Academic Press, New York, pp. 889-894.

Bonisch, M. P.; M. Huss; S. Lauber and U. Kulozik, 2007,

Fraser, J. E. and G. F. Bickerstaff, 1997, Entrapment in

Yoghurt gel formation by means of enzymatic protein

calcium

-340-

alginate.

In:

Methods

in

Biotechnology:

Jordan Journal of Agricultural Sciences, Volume 8, No.3 2012

Immobilization of enzymes and Cells. Edited by

Sci. Animal Resources, V. 23, No. 4, pp. 350-355.

Bickerstaff, G. F., Humana Press, Totowa, New Jersey, pp.

Motoki, M. and K. Seguro, 1998, Transglutaminase and its use

61-66.

for food processing. Trends Food Sci. Technol., V. 9, pp.

Guzman-Gonzalez, M.; F. Morais; M. Ramos and L. Amigo,

204-210.

1999, Influence of skimmed milk concentrate replacement

Nelson, J.A. and G.M. Trout, 1964, Judging dairy Products.

by dry dairy products in a low fat set-type yogurt model

The Olsen Publishing Co. Milwakee, Wis. 53212. U.S.A.

system. 1: Use of whey protein concentrates, milk protein

Ozer, B. H.; A. Kirmaci; S. Oztekin; A. Hayaloglu and M.

concentrates and skimmed milk powder. J. Sci. Food

Atamer, 2007, Incorporation of microbial transglutaminase

Agric., V. 79, pp. 1117-1122.

into non-fat yogurt production. Int. Dairy J., V. 17, No. 3,

Harwalkar, V.R. and M. Kalab, 1983, Susceptibility of Yogurt

pp. 199-207.

to syneresis: Comparison of centrifugation and drainage

Ozer, B. H.; R. K. Robinson; A. S. Garndison and A. Bell,

methods. Milchwissnschaft, V. 38, pp. 517-522.

1998, Gelation properties of milk concentrated by different

Han, X.; J. K. Pfeifer; R.H. Lincourt and J.M. Schuerman,

techniques. Int. Dairy J., V. 8, No. 9, pp. 793-799.

2003, Process for Making a Cheese Product Using

SAS, 2001, SAS User Guide for Personal Computer Release 6-

Transglutaminase. U.S. Patent No. 0054069A1.

18.

IDF/ISO Standard, 1991, Yogurt determination of titratable

Schorch, C.; H. Carrie and I. T. Norton, 2000, Cross-linking

acidity, (C.F. Lorenzen, et al., 2002).

casein micelles by a microbial transglutaminase : Influence

Jaros, D.; C. Heidig and H. Rohm, 2007, Enzymatic

of cross-links in acid-induced gelation. Int. Dairy J., V.

modification through microbial transglutaminase enhances

10, No. 8, pp. 529-539.

the viscosity of stirred yogurt. J. Texture Studies, V. 38,

Seguro, K.; Y. Kumazawa; C. Kuraishi; H. Sakamoto and M.

pp. 179-198.
Kuraishi,

C.;

Motoki, 1996, The - (-glutamyl) lysine moiety in cross

K.

Yamazaki

and

Y.

Susa,

2001,

linked casein is an available source of lysine for rats. J.

Transglutaminase: its utilization in food industry. Food

Nutr., V. 126, No.10, pp. 2557-2562.

Rev. Int. V. 17, No. 2, pp. 221-246.

Shah, N. P., 2003, Yogurt: The product and its manufacture,

Laemmli, U. K., 1970, Cleavage of structural protein during

in: Encyclopedia of Food Science and Nutrition, edited by

the assembly of head of bacteriology T4. Nature, V. 227,

Caballero, B.; L. C. Trugo & P. M. Finglas, V. 10, 2nd ed.,

pp. 680-685.

Academic Press. Amsterdam, UK, pp. 62526259.

Lorenzen, P.C.; H. Neve; A. Mautner and E. Schlimme, 2002,

Tamime, A., 2006, Fermented Milks 1st ed., Blackwell

Effect of enzymatic cross-linking of milk proteins on

Science Ltd., a Black- well publishing Co., Ayr., UK, pp.

54-71.

functional properties of set-style yogurt. Int. J. Dairy

Tamime, A. and R. K. Robinson, 1999, Yogurt Science and

Technol., V. 55, No. 3, pp. 152157.

Technology 2nd ed., Wood Head Publishing. London, UK.

Metwally, A. M., 2007, Effect of enzymatic crosslinking of

milk proteins on properties of ice cream with different

Zhu, Y.; A. Rinzema; J. Tramper and J. Bol, 1995, Microbial

composition. Int. J. Food Sci. Technol., V. 42, No. 8, pp.

transglutaminase a review of its production and

939-947.

application

in

food

processing.

Appl.

Biotechnol., V. 44, No. 3-4, pp. 277-282.

Moon, J. H. and Y. H. Hong, 2003, Electron microstructure

property of transglutaminase added milk. Korean J. Food

-341-

Microbiol.

Improvement of Yogurt

Waleed Ahmed Mahmood and Nawal Hurmiz Sebo

* **

) (yogurt
.Streptoverticillium mobaraense
.
4 75/.

. 45 / 3
. " .
"
.
) (SDS-PAGE


.
: .

____________________________________________
(1 .

(2 .
2011/6/15 .2012/3/5

-342-