Expression, purification, and refolding of diverse class IB hydrophobins.

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Tytuł:: Expression, purification, and refolding of diverse class IB hydrophobins.
Autorzy:: Kenward C; Department of Biochemistry & Molecular Biology, Dalhousie University, Halifax, NS, Canada.
Vergunst KL; Department of Biochemistry & Molecular Biology, Dalhousie University, Halifax, NS, Canada.
Langelaan DN; Department of Biochemistry & Molecular Biology, Dalhousie University, Halifax, NS, Canada. Electronic address: .
Źródło:: Protein expression and purification [Protein Expr Purif] 2020 Dec; Vol. 176, pp. 105732. Date of Electronic Publication: 2020 Aug 28.
Typ publikacji:: Journal Article; Research Support, Non-U.S. Gov't
Język:: English
Imprint Name(s):: Publication: Orlando, FL : Academic Press
Original Publication: San Diego : Academic Press, c1990-
MeSH Terms:: Fungal Proteins*/biosynthesis
Fungal Proteins*/chemistry
Fungal Proteins*/genetics
Fungal Proteins*/isolation & purification
Gene Expression*
Protein Refolding*
Basidiomycota/*genetics
Basidiomycota/metabolism ; Escherichia coli/genetics ; Escherichia coli/metabolism ; Recombinant Proteins/biosynthesis ; Recombinant Proteins/chemistry ; Recombinant Proteins/genetics ; Recombinant Proteins/isolation & purification
Contributed Indexing:: Keywords: Fungi; Glutathione redox buffer; Hydrophobin; Nuclear magnetic resonance spectroscopy; Protein refolding; Recombinant protein expression
Substance Nomenclature:: 0 (Fungal Proteins)
0 (Recombinant Proteins)
Entry Date(s):: Date Created: 20200901 Date Completed: 20210128 Latest Revision: 20210128
Update Code:: 20240105
DOI:: 10.1016/j.pep.2020.105732
PMID:: 32866612
: Czasopismo naukowe

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Hydrophobins are low molecular weight proteins secreted by fungi that are extremely surface-active and able to self-assemble into larger structures. Due to their unusual biochemical properties, hydrophobins are an attractive target for commercial applications such as drug emulsification and surface modification. When produced in E. coli, hydrophobins are often not soluble and need to be refolded. In this work we use SHuffle T7 Express E. coli coupled with glutathione redox buffers to produce and refold four distinct class IB hydrophobins that originate from Phanerochaete carnosa (PC1), Wallemia ichthyophaga (WI1), Serpula lacrymans (SL1), and Schizophyllum commune (SC16). Proper refolding and function of these purified hydrophobins was confirmed using nuclear magnetic resonance spectroscopy and thioflavin T assays. These results indicate that class IB hydrophobins can be consistently produced and purified from E. coli, aiding future structural and biochemical studies that require highly pure hydrophobins.
(Copyright © 2020 Elsevier Inc. All rights reserved.)

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