Document Details

Document Type : Thesis 
Document Title :
E. COLI EXPRESSION AND PHYSIOCHEMICAL PROPERTIES OF THERMOSTABLE LACTATE DEHYDROGENASE FROM A NEWLY IDENTIFIED STRAIN OF GEOBACILLUS THERMODENITRIFICANS
تعبير بكتيريا القولون والخصائص الفسيولوجية الكيميائية لإنزيم ديهيدروجينيز اللاكتات الثابت حراريًا من سلالة حديثة التعرف لجيوباسيلوس ثيرمودينايتريفيكنس
 
Subject : Faculty of Science 
Document Language : Arabic 
Abstract : Lactate Dehydrogenase (LDH) is an enzyme of glycolytic pathway, ubiquitously found in living organisms. Increased glycolysis and LDH activity are associated with many pathogenic conditions including inflammation and cancer, making the enzyme a suitable drug target. Studies on the conserved structural and functional domains of LDH from various species can reveal novel inhibitory molecules. Our study describes, E. coli production and characterization of a moderately thermostable LDH (LDH-GT) from Geobacillus thermodenitrificans DSM-465. In silico 3D model of recombinant enzyme and molecular docking with a set of potential inhibitors are also described. The recombinant enzyme was overexpressed in E. coli and purified to electrophoretic homogeneity. The molecular weight of enzyme determined by MALDI-TOF was 34798.96 Da. It exhibited maximum activity at 65°C and pH 7.5 with KM value for pyruvate as 45 µM. LDH-GT and human LDH-A have only 35.6% identity in the amino acid sequence. Contrary to that, a comparison by in silico structural alignment reveals that LDH-GT monomer has about 80% identity to that of truncated LDH-A. The amino acids ‘GEHGD’ as well as the active site His179 and His193 are conserved. Docking studies have shown the binding free energy changes of potential inhibitors with LDH-A and LDH-GT ranging -407.11 kJ mole-1 to -127.31 kJ mole-1. Highlighting the conserved structural and functional domains of LDH from two entirely different species, study has graded potential inhibitory molecules on the basis of their binding affinities that can be applied for in vivo anticancer studies. 
Supervisor : Dr. Muhammad Shahid Nadeem 
Thesis Type : Master Thesis 
Publishing Year : 1440 AH
2019 AD 
Co-Supervisor : Dr. Mustafa Adnan Zeyadi 
Added Date : Monday, April 22, 2019 

Researchers

Researcher Name (Arabic)Researcher Name (English)Researcher TypeDr GradeEmail
عبدالعزيز عوض المالكيAlmalki, Abdulaziz AwadResearcherMaster 

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