Please use this identifier to cite or link to this item: https://hdl.handle.net/10356/39819
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dc.contributor.authorYaohari, Hazarki.-
dc.date.accessioned2010-06-04T06:57:00Z-
dc.date.available2010-06-04T06:57:00Z-
dc.date.copyright2010en_US
dc.date.issued2010-
dc.identifier.urihttp://hdl.handle.net/10356/39819-
dc.description.abstractThis report summarizes the study of tobacco etch virus protease (TEVp) cleavage efficiency to enhance the productivity of antimicrobial peptide human beta-defensin (HBD) 25, a promising new class of antibiotics. Four parameters were investigated: salt concentration, substrate to enzyme ratio, cleavage time and pH. To avoid antimicrobial activity towards the host, the HBD 25 was expressed as a soluble fusion with maltose-binding protein (MBP) in Escherichia coli. Luria-Bertani (LB) was used as the culture medium for E. coli BL21 (DE3). The MBP-HBD25 fusion protein was then cleaved by the TEVp digestion enzyme at room temperature (25°C) in various conditions. The highest yield of HBD25 target peptide was obtained after 6 h of cleavage when 50 mM Tris-HCl, 2 M urea (pH = 8) digestion buffer was used, with 5:1 substrate to enzyme weight ratio. An HBD25 concentration of 0.032 g/l in the cleavage product solution was achieved with the mentioned cleavage condition. The cleavage recovery and yield were 83% and 11% (g HBD25/g substrate), respectively. Importantly, HBD25 yield was decreased by increasing salt (NaCl) concentration.en_US
dc.format.extent47 p.en_US
dc.language.isoenen_US
dc.rightsNanyang Technological University-
dc.subjectDRNTU::Engineering::Chemical engineering::Biotechnological productionen_US
dc.titleOptimization of tobacco etch virus (TEV) protease cleavage in fusion proteins.en_US
dc.typeFinal Year Project (FYP)en_US
dc.contributor.supervisorSusanna Leong Su Janen_US
dc.contributor.schoolSchool of Chemical and Biomedical Engineeringen_US
dc.description.degreeBachelor of Engineering (Chemical and Biomolecular Engineering)en_US
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Appears in Collections:SCBE Student Reports (FYP/IA/PA/PI)
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