Pengaruh Jumlah Inokulum Sel Inang Bakteri E.coli BL21 (DE3) pLysS dan Waktu Overekspresi pada Produksi Protein Rekombinan Fim-C Salmonella typhi
Recombinant protein Fim-C S.typhi is a potential protein that can be used as an alternative typhoid vaccine and produced on a large scale. However, before entering into a large-scale stage, laboratory optimum data on the factors that affect the production process of Fim-C Salmonella typhi proteins a...
| Main Authors: | , , , , , |
|---|---|
| Format: | Article |
| Language: | Indonesian |
| Published: |
Syarif Hidayatullah Jakarta State Islamic University
2018-11-01
|
| Series: | Jurnal Kimia Valensi |
| Subjects: | |
| Online Access: | http://journal.uinjkt.ac.id/index.php/valensi/article/view/8077 |
| _version_ | 1828437263085731840 |
|---|---|
| author | Muktiningsih Nurjayadi Izzatul Ilma Chairinnisa Geta Putri Mentari Dudi Hardianto Asri Sulfianti Kurnia Agustini |
| author_facet | Muktiningsih Nurjayadi Izzatul Ilma Chairinnisa Geta Putri Mentari Dudi Hardianto Asri Sulfianti Kurnia Agustini |
| author_sort | Muktiningsih Nurjayadi |
| collection | DOAJ |
| description | Recombinant protein Fim-C S.typhi is a potential protein that can be used as an alternative typhoid vaccine and produced on a large scale. However, before entering into a large-scale stage, laboratory optimum data on the factors that affect the production process of Fim-C Salmonella typhi proteins are required. This study aims to obtain information regarding the effect of host cell number E.coli BL21 (DE3) pLysS and overexpression time on production of recombinant protein Fim-C Salmonella typhi as the basis for developing candidate of typhoid vaccine. The optimization process of overexpression was done by adding 0.5 mM IPTG (Isopropyl-β-D-thiogalactopyranoside) inducer into bacterial cultures of 2%, 4%, and 6% with 4, 5, and 6 hours over expression. The measurement of the concentration of Fim-C recombinant protein extracted samples were done by a spectrophotometric method used BCA Kit Assay Thermo ScientificTM with wavelength 590nm. The characterization of those protein extracts was performed using SDS-PAGE method. The results from the study concluded that the number of 4% E.coli bacterial cells and four-hour overexpression time are the optimum condition of Fim- C Salmonella typhi characterized by the presence of high-intensity bands at ± 31 kDa. |
| first_indexed | 2024-12-10T19:42:01Z |
| format | Article |
| id | doaj.art-aa2963256fb3487fa4a41e40d1b4c261 |
| institution | Directory Open Access Journal |
| issn | 2460-6065 2548-3013 |
| language | Indonesian |
| last_indexed | 2024-12-10T19:42:01Z |
| publishDate | 2018-11-01 |
| publisher | Syarif Hidayatullah Jakarta State Islamic University |
| record_format | Article |
| series | Jurnal Kimia Valensi |
| spelling | doaj.art-aa2963256fb3487fa4a41e40d1b4c2612022-12-22T01:35:59ZindSyarif Hidayatullah Jakarta State Islamic UniversityJurnal Kimia Valensi2460-60652548-30132018-11-01429810610.15408/jkv.v4i2.80774962Pengaruh Jumlah Inokulum Sel Inang Bakteri E.coli BL21 (DE3) pLysS dan Waktu Overekspresi pada Produksi Protein Rekombinan Fim-C Salmonella typhiMuktiningsih Nurjayadi0Izzatul Ilma Chairinnisa1Geta Putri Mentari2Dudi Hardianto3Asri Sulfianti4Kurnia Agustini5Program Studi Kimia, Fakultas Matematika dan Ilmu Pengetahuan Alam, Universitas Negeri Jakarta, Gedung KH. Asj’arie Lantai 6, Jl. Rawamangun Muka, Jakarta Timur 13220, Jakarta, IndonesiaProgram Studi Kimia, Fakultas Matematika dan Ilmu Pengetahuan Alam, Universitas Negeri Jakarta, Gedung KH. Asj’arie Lantai 6, Jl. Rawamangun Muka, Jakarta Timur 13220, Jakarta, IndonesiaProgram Studi Kimia, Fakultas Matematika dan Ilmu Pengetahuan Alam, Universitas Negeri Jakarta, Gedung KH. Asj’arie Lantai 6, Jl. Rawamangun Muka, Jakarta Timur 13220, Jakarta, IndonesiaLaboratorium Pengembangan Teknologi Industri Agro dan Biomedika (LAPTIAB), Badan Pengkajian dan Penerapan Teknologi (BPPT), Serpong, Tangerang Selatan, Banten, IndonesiaLaboratorium Pengembangan Teknologi Industri Agro dan Biomedika (LAPTIAB), Badan Pengkajian dan Penerapan Teknologi (BPPT), Serpong, Tangerang Selatan, Banten, IndonesiaLaboratorium Pengembangan Teknologi Industri Agro dan Biomedika (LAPTIAB), Badan Pengkajian dan Penerapan Teknologi (BPPT), Serpong, Tangerang Selatan, Banten, IndonesiaRecombinant protein Fim-C S.typhi is a potential protein that can be used as an alternative typhoid vaccine and produced on a large scale. However, before entering into a large-scale stage, laboratory optimum data on the factors that affect the production process of Fim-C Salmonella typhi proteins are required. This study aims to obtain information regarding the effect of host cell number E.coli BL21 (DE3) pLysS and overexpression time on production of recombinant protein Fim-C Salmonella typhi as the basis for developing candidate of typhoid vaccine. The optimization process of overexpression was done by adding 0.5 mM IPTG (Isopropyl-β-D-thiogalactopyranoside) inducer into bacterial cultures of 2%, 4%, and 6% with 4, 5, and 6 hours over expression. The measurement of the concentration of Fim-C recombinant protein extracted samples were done by a spectrophotometric method used BCA Kit Assay Thermo ScientificTM with wavelength 590nm. The characterization of those protein extracts was performed using SDS-PAGE method. The results from the study concluded that the number of 4% E.coli bacterial cells and four-hour overexpression time are the optimum condition of Fim- C Salmonella typhi characterized by the presence of high-intensity bands at ± 31 kDa.http://journal.uinjkt.ac.id/index.php/valensi/article/view/8077recombinant protein fim-c salmonella typhi, salmonella typhi, overexpression |
| spellingShingle | Muktiningsih Nurjayadi Izzatul Ilma Chairinnisa Geta Putri Mentari Dudi Hardianto Asri Sulfianti Kurnia Agustini Pengaruh Jumlah Inokulum Sel Inang Bakteri E.coli BL21 (DE3) pLysS dan Waktu Overekspresi pada Produksi Protein Rekombinan Fim-C Salmonella typhi Jurnal Kimia Valensi recombinant protein fim-c salmonella typhi, salmonella typhi, overexpression |
| title | Pengaruh Jumlah Inokulum Sel Inang Bakteri E.coli BL21 (DE3) pLysS dan Waktu Overekspresi pada Produksi Protein Rekombinan Fim-C Salmonella typhi |
| title_full | Pengaruh Jumlah Inokulum Sel Inang Bakteri E.coli BL21 (DE3) pLysS dan Waktu Overekspresi pada Produksi Protein Rekombinan Fim-C Salmonella typhi |
| title_fullStr | Pengaruh Jumlah Inokulum Sel Inang Bakteri E.coli BL21 (DE3) pLysS dan Waktu Overekspresi pada Produksi Protein Rekombinan Fim-C Salmonella typhi |
| title_full_unstemmed | Pengaruh Jumlah Inokulum Sel Inang Bakteri E.coli BL21 (DE3) pLysS dan Waktu Overekspresi pada Produksi Protein Rekombinan Fim-C Salmonella typhi |
| title_short | Pengaruh Jumlah Inokulum Sel Inang Bakteri E.coli BL21 (DE3) pLysS dan Waktu Overekspresi pada Produksi Protein Rekombinan Fim-C Salmonella typhi |
| title_sort | pengaruh jumlah inokulum sel inang bakteri e coli bl21 de3 plyss dan waktu overekspresi pada produksi protein rekombinan fim c salmonella typhi |
| topic | recombinant protein fim-c salmonella typhi, salmonella typhi, overexpression |
| url | http://journal.uinjkt.ac.id/index.php/valensi/article/view/8077 |
| work_keys_str_mv | AT muktiningsihnurjayadi pengaruhjumlahinokulumselinangbakteriecolibl21de3plyssdanwaktuoverekspresipadaproduksiproteinrekombinanfimcsalmonellatyphi AT izzatulilmachairinnisa pengaruhjumlahinokulumselinangbakteriecolibl21de3plyssdanwaktuoverekspresipadaproduksiproteinrekombinanfimcsalmonellatyphi AT getaputrimentari pengaruhjumlahinokulumselinangbakteriecolibl21de3plyssdanwaktuoverekspresipadaproduksiproteinrekombinanfimcsalmonellatyphi AT dudihardianto pengaruhjumlahinokulumselinangbakteriecolibl21de3plyssdanwaktuoverekspresipadaproduksiproteinrekombinanfimcsalmonellatyphi AT asrisulfianti pengaruhjumlahinokulumselinangbakteriecolibl21de3plyssdanwaktuoverekspresipadaproduksiproteinrekombinanfimcsalmonellatyphi AT kurniaagustini pengaruhjumlahinokulumselinangbakteriecolibl21de3plyssdanwaktuoverekspresipadaproduksiproteinrekombinanfimcsalmonellatyphi |