Class I hydrophobin fusion with cellulose binding domain for its soluble expression and facile purification

Puspitasari, Nathania and Lee, Cheng-Kang (2021) Class I hydrophobin fusion with cellulose binding domain for its soluble expression and facile purification. Class I hydrophobin fusion with cellulose binding domain for its soluble expression and facile purification, 193 (116135). pp. 38-43. ISSN pISSN: 0141-8130, eISSN:1879-0003, Jurnal Internasional Bereputasi SJR(2020): 0.93, Q1, H-Index: 144

[thumbnail of Class I hydrophobin fusion with cellulose binding domain for its soluble expression and facile purification] Text (Class I hydrophobin fusion with cellulose binding domain for its soluble expression and facile purification)
1-Class_I_hydrophobin_fusion_with_(Nathania).pdf

Download (2MB)
[thumbnail of Class I hydrophobin fusion with cellulose binding domain for its soluble expression and facile purification_peer_review_] Text (Class I hydrophobin fusion with cellulose binding domain for its soluble expression and facile purification_peer_review_)
1-R1&2-Class_I_hydrophobin_fusion_.pdf

Download (1MB)
[thumbnail of Class I hydrophobin fusion with cellulose binding domain for its soluble expression and facile purification] Text (Class I hydrophobin fusion with cellulose binding domain for its soluble expression and facile purification)
1-Class_I_hydrophobin_fusion_with_hasil cek similarity.pdf

Download (2MB)

Abstract

Hydrophobins, highly surface-active proteins, have the ability to reverse surface hydrophobicity through selfassembly at the hydrophilic-hydrophobic interfaces. Their unique structure and interfacial activity lead hydrophobins to have potential applications on surface functional modifications. However, class I hydrophobins are prone to self-assemble into highly insoluble amyloid-like rodlets structure. Recombinant hydrophobins could be produced by Escherichia coli but generally as an insoluble inclusion body. To overcome this insoluble expression limitation, cellulose-binding domain (CBD) from Clostridium hermocellum was fused to the N-terminal of class I hydrophobin HGFI to enhance its soluble expression in E. coli. Approximately, 94% of expressed CBD fused HGFI (CBD-HGFI) was found as soluble protein. The fused CBD could also bind specifically onto bacterial cellulose (BC) nanofibrils produced by Komagataeibacter xylinus to facilitate rapid isolation and purification of HGFI from crude extract. Lysostaphin (Lst), known as GlyGly endopeptidase could successfully cleave the flexible linker (GGGGS)2 between CBD and HGFI to recover HGFI from BC-bound CBD-HGFI. CBD-HGFI purified by immobilized metalchelated affinity chromatography (IMAC) and Lst cleaved BC-CBD-HGFI still retained interfacial activity of hydrophobin and its effect on accelerating PETase hydrolysis against poly(ethylene terephthalate) (PET) fiber.

Item Type: Article
Additional Information: Jurnal Internasional Bereputasi SJR(2020): 0.93, Q1, H-Index: 144
Uncontrolled Keywords: Hydrophobin Cellulose binding domain Protein fusion PET hydrolysis
Subjects: Engineering > Chemical Engineering
Divisions: Journal Publication
Depositing User: F.X. Hadi
Date Deposited: 19 Aug 2022 05:19
Last Modified: 13 Sep 2022 04:18
URI: http://repository.ukwms.ac.id/id/eprint/31910

Actions (login required)

View Item View Item