Produksi aktinidin rekombinan yang diekspresikan pada khamir = Saccharomyces
Actinidin is a cysteine protease found kiwi fruit (Actinidia chinensis). In previous study, variants of actinidin - encoding gene has been cloned in yeast Saccharomyces cerevisiae to drive the expression of the gene and direct the protein to extracellular space. It had also been indicated that the r...
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[Yogyakarta] : Universitas Gadjah Mada
2005
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| _version_ | 1826031111967342592 |
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| author | Perpustakaan UGM, i-lib |
| author_facet | Perpustakaan UGM, i-lib |
| author_sort | Perpustakaan UGM, i-lib |
| collection | UGM |
| description | Actinidin is a cysteine protease found kiwi fruit (Actinidia chinensis). In previous study, variants of actinidin - encoding gene has been cloned in yeast Saccharomyces cerevisiae to drive the expression of the gene and direct the protein to extracellular space. It had also been indicated that the result of expression and secretion of recombinant actinidin in S. cerevisiae strain BF 307 - 10 carrying recombinant plasmid (pYSV9-R1) using 500 ml volume was very low. Therefore, the objective of this study was to increase the amount of recombinant actinidin produced in Saccharomyces cerevisiae .
Transformant was obtained from previous study. The expression of recombinant actinidin protein was done by cultivating transformant (S. cerevisiae strain BF 307 - 10 carrying recombinant plasmid pYSV9-R1) in 5 x 500 ml of YEPD medium. Supernatant protein was harvested at the exponential and stationary growth phase. Supernatant protein was fractionated by successive precipitation level at saturation of 30, 50, 70, and 90 % of ammonium sulfate. The results of fractionation were then dialysed and freeze-dried.
The results showed that 5 x 500 ml volume of medium produced larger amount of protein. Production of recombinant actinidin was acquired at exponential growth phase. However, at stationery growth phase, no recombinant actinidin was detected. Immunoblotting analysis demonstrated that recombinant actinidin was detected in the sample of pYSV9-R1 transformant precipitated by ammonium sulfate to 30 % saturation. The size of actinidin detected was 27,4 kD. Protein fraction containing recombinant actinidin also showed a protease activity.
Keywords: recombinant actinidin, expression, secretion, Saccharomyces
cerevisiae, incubation time, immunoblotting |
| first_indexed | 2024-03-05T22:52:16Z |
| format | Article |
| id | oai:generic.eprints.org:17567 |
| institution | Universiti Gadjah Mada |
| last_indexed | 2024-03-13T18:33:17Z |
| publishDate | 2005 |
| publisher | [Yogyakarta] : Universitas Gadjah Mada |
| record_format | dspace |
| spelling | oai:generic.eprints.org:175672014-06-18T00:27:21Z https://repository.ugm.ac.id/17567/ Produksi aktinidin rekombinan yang diekspresikan pada khamir = Saccharomyces Perpustakaan UGM, i-lib Jurnal i-lib UGM Actinidin is a cysteine protease found kiwi fruit (Actinidia chinensis). In previous study, variants of actinidin - encoding gene has been cloned in yeast Saccharomyces cerevisiae to drive the expression of the gene and direct the protein to extracellular space. It had also been indicated that the result of expression and secretion of recombinant actinidin in S. cerevisiae strain BF 307 - 10 carrying recombinant plasmid (pYSV9-R1) using 500 ml volume was very low. Therefore, the objective of this study was to increase the amount of recombinant actinidin produced in Saccharomyces cerevisiae . Transformant was obtained from previous study. The expression of recombinant actinidin protein was done by cultivating transformant (S. cerevisiae strain BF 307 - 10 carrying recombinant plasmid pYSV9-R1) in 5 x 500 ml of YEPD medium. Supernatant protein was harvested at the exponential and stationary growth phase. Supernatant protein was fractionated by successive precipitation level at saturation of 30, 50, 70, and 90 % of ammonium sulfate. The results of fractionation were then dialysed and freeze-dried. The results showed that 5 x 500 ml volume of medium produced larger amount of protein. Production of recombinant actinidin was acquired at exponential growth phase. However, at stationery growth phase, no recombinant actinidin was detected. Immunoblotting analysis demonstrated that recombinant actinidin was detected in the sample of pYSV9-R1 transformant precipitated by ammonium sulfate to 30 % saturation. The size of actinidin detected was 27,4 kD. Protein fraction containing recombinant actinidin also showed a protease activity. Keywords: recombinant actinidin, expression, secretion, Saccharomyces cerevisiae, incubation time, immunoblotting [Yogyakarta] : Universitas Gadjah Mada 2005 Article NonPeerReviewed Perpustakaan UGM, i-lib (2005) Produksi aktinidin rekombinan yang diekspresikan pada khamir = Saccharomyces. Jurnal i-lib UGM. http://i-lib.ugm.ac.id/jurnal/download.php?dataId=326 |
| spellingShingle | Jurnal i-lib UGM Perpustakaan UGM, i-lib Produksi aktinidin rekombinan yang diekspresikan pada khamir = Saccharomyces |
| title | Produksi aktinidin rekombinan yang diekspresikan pada khamir = Saccharomyces |
| title_full | Produksi aktinidin rekombinan yang diekspresikan pada khamir = Saccharomyces |
| title_fullStr | Produksi aktinidin rekombinan yang diekspresikan pada khamir = Saccharomyces |
| title_full_unstemmed | Produksi aktinidin rekombinan yang diekspresikan pada khamir = Saccharomyces |
| title_short | Produksi aktinidin rekombinan yang diekspresikan pada khamir = Saccharomyces |
| title_sort | produksi aktinidin rekombinan yang diekspresikan pada khamir saccharomyces |
| topic | Jurnal i-lib UGM |
| work_keys_str_mv | AT perpustakaanugmilib produksiaktinidinrekombinanyangdiekspresikanpadakhamirsaccharomyces |