Document Details

Document Type : Thesis 
Document Title :
RECOMBINANT PRODUCTION AND KINETIC PROPERTIES OF L-ASPARAGINASE FROM GEOBACILLUS DSM-465
الانتاج المطعم والخصائص الحركية لأنزيم ل-اسباراجينيز من جيوباسيلس DSM-465
 
Subject : Faculty of Science 
Document Language : Arabic 
Abstract : have been widely used in various industries including the pharmaceutical industry. L-asparaginase is an enzyme responsible to convert L-Asparagine to L-Aspartic acid and ammonia in a non-reversible reaction. The enzyme is universally found in animals, microbes, and plants. The enzyme from bacteria has been widely applied for the treatment of juvenile lymphomas and leukaemia. However, the enzyme often exhibits some activity against L-Glutaminase which can cause serious side-effects. The variation in enzyme’s affinity with glutamine triggers a research for very asparagine specific and highly stable enzyme. In this investigation the gene consisting of 972 bp coding for 323 amino acids was PCR amplified from the genomic DNA of Geobacillus thermodenitrificans DSM 465. The gene was T/A cloned in plasmid pTZ57RT, the gene fragment was further sub-cloned in pET22b expression vector. Recombinant of L-asparaginase was produced in E. coli strain BL21 (DE3) RIL codon plus as recombinant protein under 0.3 mM IPTG at 30◦C. The enzyme was purified by ion-exchange chromatography based on DEAE-Sephadex colum. The purified enzyme exhibited a molecular weight of about 36 kDa on SDS-PAGE. Specific activity of purified recombinant asparaginase was 1650 U per mg of protein with about 5% L-Glutaminase activity. Optimal enzyme activity was found at 75⁰C and pH 9. It exhibited a KM value of 5.9mM for L-asparagine. The enzyme was found to exist as a homotetramer by in silico studies, with estimated MW of 140 kDa ish. The enzyme produced and characterized in the present study offers a potential candidate for the treatment of leukaemia. 
Supervisor : Dr. Muhammad Shahid Nadeem 
Thesis Type : Master Thesis 
Publishing Year : 1441 AH
2019 AD 
Co-Supervisor : Dr. Mustafa Adnan Zeyadi 
Added Date : Wednesday, December 4, 2019 

Researchers

Researcher Name (Arabic)Researcher Name (English)Researcher TypeDr GradeEmail
حامد محمد السلميAlsulami, Hamed MohammedResearcherMaster 

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