HO CHI MINH UNIVERSITY OF TECHNOLOGY AND EDUCATION
FACULTY OF INTERNATIONAL EDUCATION

GRADUATION PROJECT
Thesis code: 2023 – 19116032

Students: Ho Ngoc Tram, ID: 19116032
Nguyen Tran Gia Vui, ID: 19116038
Major: Food Technology
Supervisors: Pham Thi Hoan, Ph.D.
Nguyen Huu Khang, B.E.

Ho Chi Minh City, January 2024

THE SOCIALIST REPUBLIC OF VIETNAM
Independence – Freedom– Happiness


GRADUATION THESIS ASSIGNMENT
Student name: Ho Ngoc Tram

Student ID: 19116032

Student name: Nguyen Tran Gia Vui

Student ID: 19116038

Major: Food Technology

Class: 19116 CLA

Supervisor 1: Pham Thi Hoan, Ph.D.

Email: [email protected]

Supervisor 2: Nguyen Huu Khang, B.E.

Email: [email protected]

Date of assignment: 21/08/2023

Date of submission: 22/01/2024

1. Thesis title: Production and evaluation of physicochemical and functional properties of
modified protein from Lima bean (Phaseolus lunatus) using Transglutaminase.
2. Thesis assignment:
 Establishing the process for recovering protein and modifying protein using
Transglutaminase.
 Investigating pH value, enzyme concentration and incubation time to protein
modification capacity.
 Evaluating the physicochemical properties (chemical composition, molecular weight,
particle size of protein) and functional properties (solubility, water and oil holding
capacity, emulsifying ability and stability, foaming capacity and stability) of modified
lima bean protein using Transglutaminase.
The content and requirements of the graduation thesis have been approved by the Chair of
the Food Technology program.
Ho Chi Minh City, 22nd January, 2024

CHAIR OF THE PROGRAM

SUPERVISOR

(Sign with full name)

(Sign with full name)

ACKNOWLEDGEMENT
First of all, we would like to express our sincere gratitude to all the professors, and
lecturers at the Ho Chi Minh City University of Technology and Education and with special
thanks to those in the Department of Food Technology, Faculty of Chemical and Food
Technology. They enthusiastically helped us as we conducted research and studies. Thanks to
the care, assistance, and dedicated guidance from our teachers, they have equipped us with a
solid foundation to confidently step forward on our academic and professional journey in the
future.
We especially, would like to extend our profound gratitude to the PhD. Pham Thi Hoan
is a lecturer in the Faculty of Chemical and Food Technology at Ho Chi Minh University of
Technology and Education and B.E Ngu
...

...
on Procedure
Prepared the Resolving Gel (12.5%) at the Positive Pole
Chemicals

Volume (μL)

Tris-HCl Buffer 1.5M pH 8.8

1500

Distilled Water

1900

72

Acrylamide Solution 30%

2500

SDS Solution 10%

60

APS Solution 10%

27

TEMED

4.5

Prepared the stacking gel (4%) at the negative pole
Chemicals

Volume (μL)

Tris-HCl Buffer pH 6.8

620

Distilled Water

1370

Acrylamide Solution 30%

500

SDS Solution 10%

25

APS Solution 10%

12.5

TEMED

2.5

Staining Solution: 10% (v/v) Acetic acid, 0.25% CBB-R250 or G250, and 50% (v/v)
Methanol.
De-staining Solution: 10% (v/v) Acetic acid, 25% (v/v) Methanol, and 65% water.
Appendix 4: Determination of soluble proteins by the Lowry method
Method used: Lowry et al (1954) method
Prepare the following chemical solutions
 Solution A: 0.4g NaOH (0.1N) and 2g Na2CO3 (2%) mixed in 100ml distilled water.
 Solution B: 0.5g CuSO4.5H2O (0.5%) mixed in 1% sodium and potassium
tartratesolution.
 Solution C: Mixture of two solutions A and B in a ratio of 100:2.
 Folin reagent diluted twice with distilled water before use.
 Bovine serum albumin 1mg/mL.

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Proceed
Experimental sample: Used a micropipette to accurately take 0.5ml of protein solution with
the appropriate concentration into a test tube, added 2 ml of solution C, shaken well, and let
stand for 1 minute. Then 0.25ml of Folin reagent was added to the mixture in the test tube,
shaken vigorously and evenly immediately, and left for 20 minutes until the mixture changed
to blue and reached maximum color intensity. Compared the color of the absorption mixture
on a UV-VIS machine at a wavelength of 750nm. Measured on the machine 3 times and take
the average value.
Control sample: 0.5ml of distilled water was put into a test tube and 2ml of solution C, the
next step was carried out as the experimental sample.
Calibration curve
The soluble protein content in the solution was also determined by the Lowry method,
using bovine serum albumin (BSA) as the standard.
Experimental sample: Used a micropipette to take exactly 0.5ml of BSA solution at the
above dilution concentration into a test tube. After that, 2ml of solution C was added to each
tube and left at room temperature for 10 minutes, then 0.25ml of Folin reagent was added and
put into the test tube. Colorimetric with 750nm wavelength.
Control sample: the test tube included 0.5 ml of distilled water and 2 ml of solution C and
the next steps are carried out for the experimental sample.

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