#TITLE_ALTERNATIVE#

• Main Author: NUR RAMDANI NIM:10714037, ANISA
• Format: Final Project
• Language: Indonesia
• Online Access: https://digilib.itb.ac.id/gdl/view/25629
• Institution: Institut Teknologi Bandung

The production rate of recombinant protein could be affected by presence of plasmid and <br /> <br /> composition of culture media. The presence of plasmid can be maintained with antibiotic <br /> <br /> selection markers on plasmid and addition of antibiotics into culture media. The regulatory <br /> <br /> agency restricted the uses of antibiotics in recombinant protein production due to safety reason. <br /> <br /> Culture media consists of several components such as peptone. Commonly used peptone is <br /> <br /> derived from animal protein, but currently soybeans derived peptone which is hydrolysed with <br /> <br /> papain is commercially available. This research was aimed to determine the effect of ampicillin <br /> <br /> and soy peptone on the production of superoxide dismutase recombinant Staphylococcus <br /> <br /> equorum (rMnSODSeq) in Escherichia coli TOP10. The rMnSODSeq encoding gene is carried by <br /> <br /> plasmid pCAD_sod_cer. The first step of this research is confirmation of recombinant E. coli that <br /> <br /> carry plasmid pCAD_sod_cer. The plasmid was isolated and confirmed by migration and restriction <br /> <br /> analysis. The confirmed E. coli was grown on Terrific Broth (TB) and Luria Bertani (LB) media using <br /> <br /> soy pepton or animal pepton with and without ampicillin. The effect of ampicillin and soy pepton <br /> <br /> on rMnSODSeq production were observed with SDS-PAGE and ImageJ software. The stability of <br /> <br /> plasmid segregation was determined by plasmid retention. Soy peptone increased rMnSODSeq <br /> <br /> production rate by 2.6 times compared to peptone from animals in TB and 1.4 times in LB with or <br /> <br /> without ampicillin. Plasmid pCAD_sod_cer has good segregation stability (> 90 %) even without <br /> <br /> ampicillin on media using soy peptone and pepton from animal. The use of ampicillin for the <br /> <br /> production of rMnSODSeq with pCAD_sod_cer can be removed and soy pepton can increase its <br /> <br /> production rate. <br /> <br /> <br />