EKSPRESI DAN KARAKTERISASI MUTAN PROTEIN DISULFIDA ISOMERASE RAGI YANG TIDAK MENGANDUNG DOMAIN B' DAN LINKER X
Protein disulfide isomerase (PDI) is a multi-functional enzyme involved in catalyzing reduction, oxidation, and isomerization reactions of disulfide bonds in protein molecules. The enzyme consist of two subunits. Each subunit contains a, b, b', a' and c domains. The b' domain and x li...
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id-itb.:718082023-02-24T09:14:08ZEKSPRESI DAN KARAKTERISASI MUTAN PROTEIN DISULFIDA ISOMERASE RAGI YANG TIDAK MENGANDUNG DOMAIN B' DAN LINKER X Prabudiansyah, Irfan Indonesia Final Project pdi1-?b‘x, Escherischia coli, yeast, insulin, b’ domain. INSTITUT TEKNOLOGI BANDUNG https://digilib.itb.ac.id/gdl/view/71808 Protein disulfide isomerase (PDI) is a multi-functional enzyme involved in catalyzing reduction, oxidation, and isomerization reactions of disulfide bonds in protein molecules. The enzyme consist of two subunits. Each subunit contains a, b, b', a' and c domains. The b' domain and x linker might play an important role in the substrate binding of PDI. The aims of this research were to characterize yeast PDI mutant without the b' domain and x linker (pdi1-?b'x). The pdi1-?b'x mutant and pdi1 was expressed in Escherischia coli BL21 (DE3). The pdi1-?b'x mutant was expressed as a protein with molecular weight of ± 45 kDa, while the molecular weight of the full length PDI1 was ± 60 kDa. The pdi1-?b'x still retained 65 % of insulin reduction activity. The purification of pdi1-?b’x using ammonium sulphate fractionation followed by anion exchange chromatography on DEAE-Sephacel revealed that the specific activity of pdi1-?b'x increased to 750,56 Unit/mg with a 15 fold degree of purity. The pdi1-?b’x mutant and full length PDI1 have optimum temperatur at 37 0C and optimum pH at 7.5. text |
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Protein disulfide isomerase (PDI) is a multi-functional enzyme involved in catalyzing reduction, oxidation, and isomerization reactions of disulfide bonds in protein molecules. The enzyme consist of two subunits. Each subunit contains a, b, b', a' and c domains. The b' domain and x linker might play an important role in the substrate binding of PDI. The aims of this research were to characterize yeast PDI mutant without the b' domain and x linker (pdi1-?b'x). The pdi1-?b'x mutant and pdi1 was expressed in Escherischia coli BL21 (DE3). The pdi1-?b'x mutant was expressed as a protein with molecular weight of ± 45 kDa, while the molecular weight of the full length PDI1 was ± 60 kDa. The pdi1-?b'x still retained 65 % of insulin reduction activity. The purification of pdi1-?b’x using ammonium sulphate fractionation followed by anion exchange chromatography on DEAE-Sephacel revealed that the specific activity of pdi1-?b'x increased to 750,56 Unit/mg with a 15 fold degree of purity. The pdi1-?b’x mutant and full length PDI1 have optimum temperatur at 37 0C and optimum pH at 7.5. |
| format |
Final Project |
| author |
Prabudiansyah, Irfan |
| spellingShingle |
Prabudiansyah, Irfan EKSPRESI DAN KARAKTERISASI MUTAN PROTEIN DISULFIDA ISOMERASE RAGI YANG TIDAK MENGANDUNG DOMAIN B' DAN LINKER X |
| author_facet |
Prabudiansyah, Irfan |
| author_sort |
Prabudiansyah, Irfan |
| title |
EKSPRESI DAN KARAKTERISASI MUTAN PROTEIN DISULFIDA ISOMERASE RAGI YANG TIDAK MENGANDUNG DOMAIN B' DAN LINKER X |
| title_short |
EKSPRESI DAN KARAKTERISASI MUTAN PROTEIN DISULFIDA ISOMERASE RAGI YANG TIDAK MENGANDUNG DOMAIN B' DAN LINKER X |
| title_full |
EKSPRESI DAN KARAKTERISASI MUTAN PROTEIN DISULFIDA ISOMERASE RAGI YANG TIDAK MENGANDUNG DOMAIN B' DAN LINKER X |
| title_fullStr |
EKSPRESI DAN KARAKTERISASI MUTAN PROTEIN DISULFIDA ISOMERASE RAGI YANG TIDAK MENGANDUNG DOMAIN B' DAN LINKER X |
| title_full_unstemmed |
EKSPRESI DAN KARAKTERISASI MUTAN PROTEIN DISULFIDA ISOMERASE RAGI YANG TIDAK MENGANDUNG DOMAIN B' DAN LINKER X |
| title_sort |
ekspresi dan karakterisasi mutan protein disulfida isomerase ragi yang tidak mengandung domain b' dan linker x |
| url |
https://digilib.itb.ac.id/gdl/view/71808 |
| _version_ |
1822006685653270528 |