Production and purification of alpha hemolysin produced from local isolate of Serratia marcescens

Dissertant

al-Ajili, Amir Qani Kazim

Thesis advisor

al-Tai, Maha Fakhri
Hamzah, Subhi Jawad

Comitee Members

al-Nida, Sad M.
Kazim, Khalid Jabir

University

University of Baghdad

Faculty

College of Science

Department

Biotechnology Department

University Country

Iraq

Degree

Master

Degree Date

2014

English Abstract

Twenty isolates that have been identified as Serratia marcescens were collected from infected urinary tract infection (UTI).

These isolates were found to produce hemolysin as indicated by clear zone around colonies grown on blood agar plates.

Isolates were screened for their production and the highest producing isolate was selected for further experiments.

Hemolysin was completely precipitated using 50-75% saturation of ammonium sulphate.

Then followed by ion exchange chromatography with DEAE-cellulose, The hemolysin was not detected in the wash step, while in elution step, only one of the two peaks showed hemolytic activity by measuring the absorbance of hemoglobin at 405 nm and as much as 29 μg/ml of hemolysin was quantified.

Finally hemolysin was completely purified to estimate the molecular weight which was shown to be 45 KD by using gel filtration chromatography using sepharose 4B, and hemolysin concentration was reached to 22 μg/ml.

Subsequently, three doses of hemolysin (1, 5 and 10)μg/ml were given orally for seven days to experimental animals (mice) in order to measure the cytogenetic effects of this toxin using three parameters, (mitotic index (MI), micronulcleus (MN) and chromosomal aberrations (CAs) in comparison with phosphate buffer saline (PBS) as a negative control and mitomycin C (MMC) as a positive control.

The results of the effects of the three doses of hemolysin on mice indicated that all doses caused a significant decrease in MI which were shown to be (1.10%, 0.71 % and 0.32%) compared with negative control (1.742%) and positive control (0.742%).

Moreover, a significant increase in the percentage of MN was observed to be (1.92%, 2.88% and 4.04%) in comparison with negative control (0.16%) and positive control (4.132%).

Also a significant increase was seen in the values of CAs which were (0.744, 1.006 and 1.480 %) compared with negative control (0.244% ) and positive control ( 1.498%).

Main Subjects

Biology

Topics

• Physiology
• Genetics

No. of Pages

104

Table of Contents

• APA
• MLA
• AMA

Language

English

Data Type

Arab Theses

Record ID

BIM-601072