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    Separation and Purification of Industrial Proteins by Chromatography and Aqueous two Phase System

    , M.Sc. Thesis Sharif University of Technology Yavari, Milad (Author) ; Vossoughi, Manouchehr (Supervisor) ; Seifkordi, Ali Akbar (Supervisor)
    Abstract
    Separation of biomolecules such as enzymes is the most important part of biotechnology process which 10-90 % of costs is for separation and purification. There are several methods for purification of bio materials and one of them is chromatography which use in enzyme purification but this method have several problem such as cost ,time consuming and scale up Because of this a suitable alternative demands for industrial separation. An alternative is aqueous two-phase systems (ATPS). In this project was studied about partial purification of alkaline protease produced by Bacillus licheniformis ATCC 21424 in aqueous two-phase system and comparison with chromatography method. an aqueous two-phase... 

    Using Vitamin B1 as a Catalyst in Performing Multi-Component Reactions and Synthesis of Affinity Column ChromatograpHy for Purification of Recombinant Proteins

    , M.Sc. Thesis Sharif University of Technology Lohi Khosroshahi, Ameneh (Author) ; Kalhor, Hamid Reza (Supervisor)
    Abstract
    Due to the importance of using pure proteins in the industry and basic research in order to study them in the structural contexts and interactions of proteins in recent years, purification methods have been extensively developed. The purification steps usually depend on the size of the protein, its physical and chemical properties, the binding affinity and the biological activity of the protein in question, In the purification process, the protein portion is separated from the non-protein portion. The biggest challenge is when we want to separate different proteins. Different chromatographic methods are used, depending on the properties of the impure proteins. One of these chromatographic... 

    Cloning and Production of the Flavin Reductase Enzyme in Order to Perform Organic Reaction and Synthesis of Cation Exchange Column for Protein Purification

    , M.Sc. Thesis Sharif University of Technology Hayati, Fatemeh (Author) ; Kalhor, Hamid Reza (Supervisor)
    Abstract
    The reductase enzymes perform an important function in the cell. These enzymes are usually coupled with other enzymes and participate in reactions such as halogenation, reduction of alkene, and carboxylic acids. These reactions are carried out by reproducing cofactors such as FADH2 and NAD+; however, there is no evidence that this enzyme alone can perform an organic reaction. In this work, we aim to examine whether recombinant flavin reductase can carry out organic reactions. In order to amplify the flavin reductase enzyme gene from the E. coli genome by PCR (Polymerase Chain Reaction) method, two synthetic primers were designed; The forward primer of the gene contained a cleavage site for... 

    Producing functional recombinant human keratinocyte growth factor in Pichia pastoris and investigating its protective role against irradiation

    , Article Enzyme and Microbial Technology ; Volume 111 , April , 2018 , Pages 12-20 ; 01410229 (ISSN) Bahadori, Z ; Kalhor, H. R ; Mowla, S. J ; Sharif University of Technology
    Elsevier Inc  2018
    Abstract
    Keratinocyte Growth Factor (KGF) is a paracrine-acting, epithelial mitogen that plays a prominent role in the regeneration of damaged epithelial tissues. In spite of different attempts to produce recombinant human KGF in many organisms, including bacteria, mammalian cells, plant cells and insect cells; production of recombinant form suffers from lower yields and recovery relative to other recombinant proteins of similar size and properties. Due to many advantages of Pichia pastoris expression systems for producing industrial enzymes and pharmaceutical proteins, in this study P. pastoris was chosen as a host for KGF expression. For preparing human KGF coding sequence, MCF-7 cell line was...