Production, purification and characterization of alginase from Bacillus circulans local isolate
Other Title(s)
إنتاج و تنقية و توصيف أنزيم الألجنيز من بكتريا Bacillus circulans المعزولة محليا
Dissertant
Thesis advisor
University
University of Baghdad
Faculty
Genetic Engineering and Biotechnology for Post Graduate Studies
University Country
Iraq
Degree
Master
Degree Date
2015
English Abstract
Fourteen bacteria isolates “ Bacillus species” were taken from Ramadi and Fallujah hospital in the period between October to December 2013, those isolated were cultured on a blood agar to test their ability to hydrolytic due to formation the inhibition zone .
Six isolate were selected to be cultured in alginate yeast extract broth for testing their efficiency to produce alginase .The efficient isolate was diagnosed depending on phenotypic, microscopic and biochemical tests to be Bacillus circulans R . The optimal conditions for alginase production by Bacillus circulans R as follows; the optimum incubation period was 24 hrs, which gave enzymatic activity (106.1U/ml) .
The optimum inoculum optimum inoculum volume was 6x107 cell/g dry weight, which gave activity (111 Unit/ml).
The optimum carbon source was sodium alginate which gave activity (212.86 U/ml), while peptone was the optimum nitrogen sources with enzymatic activity of (275 U/ml ).
The optimum pH was 7.5 which gave maximum activity (211.63 U/ml) , the shaking incubator was gave a highest production of the enzyme activity (210 U/ml). The enzyme from Bacillus circulans R was purified by several steps, Including ammonium sulfate precipitation with a saturation percentage of 50-75% which was gave specific activity (2933.15U/mg) with a fold number of (3.49) and a yield of ( 58.75%).
Ionic exchange chromatography using DEAE-Sephacel column ( 3.5 x8 cm) gave specific activity (5362.33U/mg), a fold number of (6.37) and a yield of (53.11%), then gel filtration using Sepharose 6 B column (1.5x87 cm) gave purified enzyme with specific activity of (10865U/mg), a fold number of (12.91) and a yield (28.69%).
The results of alginase characterization were: · The optimum pH for alginase activity was 7.0. · The optimum pH for enzyme stability was 7.5 . · The optimum temperature for alginase activity was 35°C. · The enzyme retained its entire activity upon 1 hour incubation at temperature 25°C . · The molecular weight of the alginase as determined by gel filtration chromatography using Sepharose 6 B gel was 40738 Dalton and by SDSelectrophoresis was49000 Daltons. The gene responsible for controlling production of alginase in Bacillus circulans R was located in the chromosomal DNA.
The alginase has ability to degrade alginate coated Pseudomonas aeruginosa when treated with it after incubated for one hour
Main Subjects
No. of Pages
108
Table of Contents
Table of contents.
Abstract.
Abstract in Arabic.
Chapter One : Introduction.
Chapter Two : Literature review.
Chapter Three : Materials and methods.
Chapter Four : Results and discussion.
Conclusions and recommendations.
References.
American Psychological Association (APA)
al-Halbusi, Rafal Ismail Ali. (2015). Production, purification and characterization of alginase from Bacillus circulans local isolate. (Master's theses Theses and Dissertations Master). University of Baghdad, Iraq
https://search.emarefa.net/detail/BIM-736084
Modern Language Association (MLA)
al-Halbusi, Rafal Ismail Ali. Production, purification and characterization of alginase from Bacillus circulans local isolate. (Master's theses Theses and Dissertations Master). University of Baghdad. (2015).
https://search.emarefa.net/detail/BIM-736084
American Medical Association (AMA)
al-Halbusi, Rafal Ismail Ali. (2015). Production, purification and characterization of alginase from Bacillus circulans local isolate. (Master's theses Theses and Dissertations Master). University of Baghdad, Iraq
https://search.emarefa.net/detail/BIM-736084
Language
English
Data Type
Arab Theses
Record ID
BIM-736084
