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...
Saved in:
| Main Author: | |
|---|---|
| Format: | Final Project |
| Language: | Indonesia |
| Online Access: | https://digilib.itb.ac.id/gdl/view/71808 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| Institution: | Institut Teknologi Bandung |
| Language: | Indonesia |
| Summary: | 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. |
|---|