Extraction and purification of Asparaginase enzyme from Pisum sativum plant and studying their cytotoxicity against L20B tumor cell line
Other Title(s)
استخلاص وتنقية إنزيم الاسباراجينيز المعزول من نبات البزاليا Pisum sativum و دراسة سميته ضد الخط السرطاني L20B
Dissertant
Thesis advisor
Jasim, Hamid M.
al-Ani, Nabil Khalaf
Comitee Members
Shamki, Abd al-Wahid J.
al-Tai, Muhammad Ibrahim Nadir Mahmud
University
University of Baghdad
Faculty
College of Science
Department
Biotechnology Department
University Country
Iraq
Degree
Master
Degree Date
2012
English Abstract
Plant samples of Pisum sativum were collected from crop fields in the Collage of Agriculture/ University of Baghdad and were classified as Pisum sativum subspp.
Jof according to their morphological characteristics.
Activity of asparaginase was detected in seeds, stems and leaves extracts.
Results showed that maximum asparaginase activity was detected in seeds extracts which was 30.0 U/mg in comparison with 26.4 and 16.1 U/mg in extracts of leaves and stems respectively.
According to these results plant seeds were used as a source for asparaginase production, characterization, and studying its antitumor activity.
Optimum conditions for the activity of crude asparaginase extracted from plants seeds were studied.
Results showed maximum activity of asparaginase was achieved when the enzyme was incubated with 200mM of asparagines in a ratio of 1:3 (V/V) at 37°C for 30 minutes in presence of 0.05 M of potassium phosphate buffer solution at pH8.
Crude asparaginase extracted from plant seeds was purified in two steps, ion exchange chromatography by DEAE-Cellulose and gel filtration chromatography by Sephadex G-200.
Specific activity of purified asparaginase was 228.8 U/mg.
Asparaginase purified from seeds extracts was then characterized.
Results of characterization showed that the molecular weight of asparaginase was 66,464 Kelo dalton, and the optimum pH for enzyme activity and stability was pH 8.5, while the optimum temperature for enzyme activity and stability was 37°C and 40°C respectively.
On the other hand the enzyme Chapter One Introduction and Literature review 6 activation energy was 6260 calories/mol, and the temperature coefficient (Q10) for asparaginase was 1.32.
Antitumor activity for the purified asparaginase was studied using L20B tumor cell line by incubation with gradual concentration of purified asparaginase for 48 hours.
Results showed that asparaginase extracted and purified from seeds of P.
sativum has inhibitory effect on L20B tumor cell line.
Main Subjects
Topics
No. of Pages
140
Table of Contents
Table of contents.
Abstract.
Abstract in Arabic.
Chapter One : Introduction and literature review.
Chapter Two : Materials and methods.
Chapter Three : Results and discussions.
Conclusions and recommendations.
References.
American Psychological Association (APA)
Khalaf, Zaynah Abd Allah. (2012). Extraction and purification of Asparaginase enzyme from Pisum sativum plant and studying their cytotoxicity against L20B tumor cell line. (Master's theses Theses and Dissertations Master). University of Baghdad, Iraq
https://search.emarefa.net/detail/BIM-601054
Modern Language Association (MLA)
Khalaf, Zaynah Abd Allah. Extraction and purification of Asparaginase enzyme from Pisum sativum plant and studying their cytotoxicity against L20B tumor cell line. (Master's theses Theses and Dissertations Master). University of Baghdad. (2012).
https://search.emarefa.net/detail/BIM-601054
American Medical Association (AMA)
Khalaf, Zaynah Abd Allah. (2012). Extraction and purification of Asparaginase enzyme from Pisum sativum plant and studying their cytotoxicity against L20B tumor cell line. (Master's theses Theses and Dissertations Master). University of Baghdad, Iraq
https://search.emarefa.net/detail/BIM-601054
Language
English
Data Type
Arab Theses
Record ID
BIM-601054
