Efficient purification of recombinant proteins using hydrophobins as tags in surfactant-based two-phase systems

Biochemistry
Markus B LinderM Penttilä

Abstract

In this work we describe the new concept of using fungal hydrophobins as efficient tags for purification of recombinant fusion proteins by aqueous two-phase separation. Hydrophobins are a group of small surface-active proteins produced by filamentous fungi. Some characteristics of hydrophobins are that they are relatively small (approximately 100 amino acids), they contain eight disulfide-forming Cys residues in a conserved pattern, and they self-assemble on interfaces. The aqueous two-phase systems studied were based on nonionic surfactants that phase-separate at certain temperatures. We show that the use of hydrophobins as tags has many advantages such as high selectivity and good yield and is technically very simple to perform. Fusion proteins with target proteins of different molecular size were compared to the corresponding free proteins using a set of different surfactants. This gave an understanding on which factors influence the separation and what rationale should be used for optimization. This unusually strong and specific interaction between polymeric surfactants and a soluble protein shows promise for new developments in interfacing proteins and nonbiological materials for other applications as well.

References

Aug 15, 1991·Analytical Biochemistry·H WalterD E Brooks
Apr 1, 1993·Journal of Biotechnology·G C TerstappenM R Kula
Jan 15, 1996·European Journal of Biochemistry·T Nakari-SetäläM Penttilä
Nov 5, 1997·Protein Expression and Purification·J NilssonP A Nygren

❮ Previous
Next ❯

Citations

Jan 30, 2013·Molecular Biotechnology·Stefan R Schmidt
Mar 19, 2013·European Journal of Pharmaceutical Sciences : Official Journal of the European Federation for Pharmaceutical Sciences·Hanna ValoTimo Laaksonen
Sep 7, 2006·Journal of the American Chemical Society·Byung-Keun OhChad A Mirkin
Mar 19, 2011·Langmuir : the ACS Journal of Surfaces and Colloids·Elka S BashevaAlex Lips
Jul 10, 2013·Applied and Environmental Microbiology·Michael LienemannMarkus B Linder
Mar 16, 2006·Proceedings of the National Academy of Sciences of the United States of America·A H Y KwanJ P Mackay
Feb 9, 2012·Trends in Biotechnology·Päivi LaaksonenMarkus B Linder
Aug 2, 2011·Journal of Controlled Release : Official Journal of the Controlled Release Society·Hanna ValoTimo Laaksonen
Oct 4, 2015·Protein Expression and Purification·Azadeh Lohrasbi-NejadSaman Hosseinkhani
Sep 19, 2007·Micron : the International Research and Review Journal for Microscopy·Margaret SundeJoel P Mackay
Apr 24, 2007·Analytica Chimica Acta·Aline Soriano LopesMarco Aurélio Zezzi Arruda
Sep 6, 2015·New Biotechnology·Angela SunHelena Nevalainen
Mar 11, 2016·ACS Applied Materials & Interfaces·Miika SoikkeliJouni Ahopelto
Dec 18, 2013·Plant Biotechnology Journal·Lauri J ReuterAnneli Ritala
Nov 10, 2015·BioMed Research International·Ghislain MoussavouYoung-Kug Choo
Mar 11, 2006·Biomolecular Engineering·Kalyani Mondal, Munishwar N Gupta
Jul 26, 2015·The Protein Journal·Mohammadreza KhalesiGuy Derdelinckx
Apr 7, 2012·Journal of Colloid and Interface Science·Nikola A AlexandrovAlex Lips
Mar 24, 2012·Protein Expression and Purification·Baolong NiuMingqiang Qiao
Oct 13, 2005·FEMS Microbiology Reviews·Markus B LinderMerja E Penttilä
Feb 13, 2008·Protein Expression and Purification·Tomi LahtinenChristian Oker-Blom
Mar 30, 2010·Protein Expression and Purification·Zefang WangMingqiang Qiao
Jun 19, 2013·European Journal of Pharmaceutics and Biopharmaceutics : Official Journal of Arbeitsgemeinschaft Für Pharmazeutische Verfahrenstechnik E.V·Ruzica KolakovicTimo Laaksonen
Jan 8, 2015·Applied Microbiology and Biotechnology·Han A B Wösten, Karin Scholtmeijer
Jun 5, 2016·Advances in Colloid and Interface Science·Weiwei Zhao, Yilin Wang
Jun 12, 2016·Microbial Cell Factories·Robert H BischofBernhard Seiboth
Jun 15, 2016·Philosophical Transactions. Series A, Mathematical, Physical, and Engineering Sciences·Ryan J MorrisCait E MacPhee
Oct 6, 2016·PloS One·Lauri ReuterJussi Joensuu
Jul 25, 2018·Current Protocols in Protein Science·Helena NevalainenValentino Setoa Junior Te'o
Jun 24, 2017·Plant Biotechnology Journal·Katri KurppaJussi J Joensuu
Sep 6, 2010·Materials·Filippo ZampieriKarin Scholtmeijer
Jan 28, 2021·Molecular and Cellular Biochemistry·Hengfang TangPeng Wang
Feb 8, 2020·Protein Expression and Purification·Rezwan SiddiqueeAnn Hau-Yu Kwan

❮ Previous
Next ❯

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.