#TITLE_ALTERNATIVE#

• Main Author: NIM : 20715308, SUSILAWATI
• Format: Theses
• Language: Indonesia
• Online Access: https://digilib.itb.ac.id/gdl/view/31160
• Institution: Institut Teknologi Bandung

Halal Peptone (HY-SOY T) is protein of soybeans that have been hydrolyzed <br /> <br /> using papaya papain enzyme become a simple peptide. The use of halal peptone <br /> <br /> for the production of recombinant protein have the advantages such as in religion, <br /> <br /> economy and there is no information on the risk of plant pathogens to human. <br /> <br /> This research was aimed to evaluate the effect halal peptone on the production of <br /> <br /> recombinant superoxide dismutase (rMnSODSeq) protein in Escherichia coli. In <br /> <br /> this research, halal peptone was used in two types of media, Luria Bertani (LB) <br /> <br /> and Terrific Broth (TB) and compared with tryptone from animal that is <br /> <br /> commonly used in Pharmaceutical Biotechnology Laboratory ITB. There are two <br /> <br /> rMnSODSeq expression systems used in this research, pCAD_sod_cer which <br /> <br /> depends on growth phase and pJExpress414_sod which depends on the addition <br /> <br /> of inducer. In the first phase of this research was confirmation of E. coli that <br /> <br /> carry each plasmid when grown in medium with halal peptone. Plasmids that <br /> <br /> isolated from each E. coli were analyzed migration profiles, size and the presence <br /> <br /> of rMnSODSeq encoding gene. The next step was evaluation of the effect of halal <br /> <br /> pepton on the recombinant E. coli growth profile by measuring the cell density at <br /> <br /> a certain time. Effect on the production and activity of rMnSODSeq was <br /> <br /> performed by comparing the intensity of the rMnSODSeq protein band on SDS- <br /> <br /> PAGE acrylamide gel and analysed by zymography. Both recombinant E. coli <br /> <br /> were positively harboring pCAD_sod_cer and pJExpress414_sod expression <br /> <br /> system. The growth of both E. coli were observed slower when halal peptone was <br /> <br /> used in LB and TB media. Mean while the production of rMnSODSeq was <br /> <br /> 33,36% more on LB medium with halal pepton using pCAD_sod_cer expression <br /> <br /> system and 31,64% using pJExpress414_sod expression system. This increase <br /> <br /> also observed 47,41% more on TB media with halal peptone using <br /> <br /> pCAD_sod_cer expression system and 64,89% using pJExpress414_sod <br /> <br /> expression system. Dismutase activity of rMnSODSeq production using TB media <br /> <br /> with halal peptone still showed positive result on zymography analysis. Halal <br /> <br /> peptone has reduced the growth of host cells but increased the production of <br /> <br /> proteins and did not affect the dismutation of rMnSODSeq activity. <br />