KAJIAN EKSPRESI PROTEIN YsdC, BSUW23_10175 DAN XynB DARI Bacillus subtilis subsp.spizizenii W23 SECARA ALAMI DAN DI DALAM SEL INANG Escherichia coli Origami
| Main Author: | Wahjudi, Mariana |
|---|---|
| Format: | Article PeerReviewed application/pdf |
| Terbitan: |
Lembaga Penelitian dan Pengabdian Kepada Masyarakat, Universitas Surabaya
, 2013
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| Subjects: | |
| Online Access: |
http://repository.ubaya.ac.id/24782/1/Wahjudi_Kajian%20Ekspresi%20Protein_Abstrak_2013.pdf http://repository.ubaya.ac.id/24782/2/Wahjudi_Kajian%20Ekspresi%20Protein_2013.pdf http://repository.ubaya.ac.id/24782/ |
Daftar Isi:
- Cellulases and xylanases are widely used in various applications such as in laundry and waste managements; and in various industries such as candy, food, drink, and pulp industries. In previous study, the genes which encoded endo-1,4-beta-glucanase (ysdC), endo-beta-1,3-1,4 glucanase (BSUW23_10175), and xylan beta-1,4- xylosidase (xynB) from Bacillus subtilis subsp. spizizenii str. W23 were cloned into broad-host-range vector pMMB67EH, in Escherichia coli Origami cells. However, these protein can be processed unproperly in Gramnegative bacteria differ than in its native Gram positive host cell. The aim of this research was to study the processing dan translocation of these native proteins in Bacillus subtilis subsp. spizizenii str. W23 and in E. coli Origami transformants cells, by in silico study dan expression study. Analyses of signal peptide, protein processing and translocation were accomplished by several softwares, i.e. PrediSi MHTMM and SignalP softwares. Expression study was performed by growing cells in Luria-Bertani (LB) broth with addition of carboxymethyl-cellulosa (CMC) or xylan as substrate and isopropyl-~-D-1-thiogalactopyranoside (IPTG) as inducer. The expressed protein was detected by sodium-dodecylsulphate-polyacrylamide gel-electrophoresis (SDS-PAGE) technique or by activity assays towards its substrates. The result showed that YsdC, BSUW23_10175 and XynB proteins were over-expressed in E. coli Origami trasformants cells. The YsdC and XynB proteins were expressed as fully active-extracellular enzyme protein, however BSUW23_10175 protein was detected as intracellular un-processed pro~in, that is differ from its native and predicted.