Document Details

Document Type : Thesis 
Document Title :
Immobilization of Horseradish Peroxidase on Natural Polymer
تحميل إنزيم بيروكسيديز الفجل على مبلمر طبيعى
 
Subject : Biochemistry department 
Document Language : Arabic 
Abstract : The immobilized enzyme has offered several advantages, such as enhanced stability, easier product recovery and purification, protection of enzymes against denaturants, proteolysis and reduced susceptibility to contamination. Therefore, the goal of this work is to immobilize horseradish peroxidase (HRP) to be compatible with its further application. The methods included partial purification of HRP using ammonium sulphate precipitation, immobilization HRP on modified wool by cyanuric chloride and characterization of soluble of immobilized enzyme. The fourier transform infrared FT-IR spectroscopy and scanning electron microscopy (SEM) were used to characterize HRP immobilization. The immobilized HRP retained 62% of initial activity after six reuses. Immobilized HRP showed broad optimum pH at 7.0 and 8.0, which was higher than that of the soluble HRP (pH 6.0). The soluble HRP had an optimum temperature of 30°C, which was shifted to 40°C for immobilized enzyme. The soluble and immobilized HRP were stable up to 30 and 40°C after incubation for 15 min. The immobilized HRP was able to react with all tested substrates and the velocity of the product for immobilized enzyme was higher than that soluble enzyme The apparent kinetic constant values (Kms) of immobilized HRP were 10 mM for guiacol and 2.5 mM for H2O2, which were higher than that of soluble HRP. The immobilized HRP was remarkably more stable against proteolysis mediated by trypsin. The immobilized HRP exhibited more resistance to heavy metal induced inhibition. The immobilized HRP was more stable to the denaturation induced by urea, Triton X-100 and isopropanol. The soluble HRP lost 46% of its activity after 16 weeks of storage, while the immobilized HRP retained 86% of its original activity at the same time. In conclusion, the immobilized HRP could be exploited for several industrial and environmental purposes. 
Supervisor : DR. Saleh Ahmed Mohamed 
Thesis Type : Master Thesis 
Publishing Year : 1433 AH
2012 AD 
Co-Supervisor : DR. Reda Mohamed El-Shishtawy 
Added Date : Tuesday, July 10, 2012 

Researchers

Researcher Name (Arabic)Researcher Name (English)Researcher TypeDr GradeEmail
أنس عابد درويشDarwish, Anas AbedResearcherMaster 

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