Jefferson, Timotius William and Pandjaitan, Maruli and Nasution, Uli Julia (2019) Cephalosporin C Acylase Characterization & Purification Using Chromatography Techniques. Bachelor thesis, Swiss German University.

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Abstract

Cephalosporin C Acylase is an enzyme that plays an important role in the one step conversion of Cephalosporin C into 7-aminocephalosporanic acid (7-ACA), a key intermediate for the synthesis of semisynthetic cephalosporin antibiotics. Purification is a method or process of removing unwanted protein molecules or impurities from protein of interest with the aim of increasing enzyme specific activity. The purification process begins by expressing Cephalosporin C Acylase using E. coli BL21 (DE3) followed by the cell lysis process. Characterization of Cephalosporin C Acylase was being examined in this study to get a profile of the enzyme before entering purification process. This study applied chromatography techniques for enzyme purification, but mainly focusing on a specific purification method, Immobilized Metal Affinity Chromatography. It is a widely used purification method to purify histidine tagged protein. Ion-exchange Chromatography was also conducted as one of the chromatography techniques for additional purification step. Purification process in this research study is capable to increase enzyme specific activity up to 4.56-fold and recover 63.9% yield. Result of protein bands analysis using SDS-PAGE techniques shown that recombinant CCA enzyme is consist of two subunits, α (25kDA) and β (58kDa) subunit. This analysis also shown that purification process is capable in removing impurities from enzyme of interest.

Item Type:	Thesis (Bachelor)
Uncontrolled Keywords:	Cephalosporin C Acylase, Protein Purification, Chromatography Techniques, Enzyme Specific Activity
Subjects:	R Medicine > R Medicine (General) > R856 Biomedical engineering
Divisions:	Faculty of Life Sciences and Technology > Department of Biomedical Engineering
Depositing User:	Adityatama Ratangga
Date Deposited:	19 May 2020 12:08
Last Modified:	08 Jun 2021 16:05
URI:	http://repository.sgu.ac.id/id/eprint/509

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