Intracellular protein production in Trichoderma reesei (Hypocrea jecorina) with hydrophobin fusion technology

Eero Mustalahti, Markku Saloheimo, Jussi Joensuu (Corresponding Author)

Research output: Contribution to journalArticleScientificpeer-review

14 Citations (Scopus)

Abstract

Insufficient accumulation and the lack of efficient purification methods are the two major bottlenecks hindering the recombinant production of many proteins. Alternative production schemes are urgently needed for proteins that remain challenging to express and purify with conventional techniques. We have found that hydrophobin fusions targeted to endoplasmic reticulum (ER) can enhance the expression of target proteins simultaneously providing means for straightforward purification. Here we show that hydrophobin fusion technology induces formation of large protein bodies in the filamentous fungus Trichoderma reesei. The fusion protein remained soluble in the ER-derived protein bodies. A simple and scalable aqueous two-phase system was demonstrated to purify the hydrophobin fusion protein GFP-HFBI from the complex intracellular extracts with a recovery of up to 62%.
Original languageEnglish
Pages (from-to)262-268
Number of pages7
JournalNew Biotechnology
Volume30
Issue number2
DOIs
Publication statusPublished - 2013
MoE publication typeA1 Journal article-refereed

Fingerprint

Hypocrea
Trichoderma
Fusion reactions
Technology
Proteins
Endoplasmic Reticulum
Purification
Fungi
Complex Mixtures
Recovery

Cite this

@article{0c07a5576ef0488ca7f4fd694e5da66d,
title = "Intracellular protein production in Trichoderma reesei (Hypocrea jecorina) with hydrophobin fusion technology",
abstract = "Insufficient accumulation and the lack of efficient purification methods are the two major bottlenecks hindering the recombinant production of many proteins. Alternative production schemes are urgently needed for proteins that remain challenging to express and purify with conventional techniques. We have found that hydrophobin fusions targeted to endoplasmic reticulum (ER) can enhance the expression of target proteins simultaneously providing means for straightforward purification. Here we show that hydrophobin fusion technology induces formation of large protein bodies in the filamentous fungus Trichoderma reesei. The fusion protein remained soluble in the ER-derived protein bodies. A simple and scalable aqueous two-phase system was demonstrated to purify the hydrophobin fusion protein GFP-HFBI from the complex intracellular extracts with a recovery of up to 62{\%}.",
author = "Eero Mustalahti and Markku Saloheimo and Jussi Joensuu",
year = "2013",
doi = "10.1016/j.nbt.2011.09.006",
language = "English",
volume = "30",
pages = "262--268",
journal = "New Biotechnology",
issn = "1871-6784",
publisher = "Elsevier",
number = "2",

}

Intracellular protein production in Trichoderma reesei (Hypocrea jecorina) with hydrophobin fusion technology. / Mustalahti, Eero; Saloheimo, Markku; Joensuu, Jussi (Corresponding Author).

In: New Biotechnology, Vol. 30, No. 2, 2013, p. 262-268.

Research output: Contribution to journalArticleScientificpeer-review

TY - JOUR

T1 - Intracellular protein production in Trichoderma reesei (Hypocrea jecorina) with hydrophobin fusion technology

AU - Mustalahti, Eero

AU - Saloheimo, Markku

AU - Joensuu, Jussi

PY - 2013

Y1 - 2013

N2 - Insufficient accumulation and the lack of efficient purification methods are the two major bottlenecks hindering the recombinant production of many proteins. Alternative production schemes are urgently needed for proteins that remain challenging to express and purify with conventional techniques. We have found that hydrophobin fusions targeted to endoplasmic reticulum (ER) can enhance the expression of target proteins simultaneously providing means for straightforward purification. Here we show that hydrophobin fusion technology induces formation of large protein bodies in the filamentous fungus Trichoderma reesei. The fusion protein remained soluble in the ER-derived protein bodies. A simple and scalable aqueous two-phase system was demonstrated to purify the hydrophobin fusion protein GFP-HFBI from the complex intracellular extracts with a recovery of up to 62%.

AB - Insufficient accumulation and the lack of efficient purification methods are the two major bottlenecks hindering the recombinant production of many proteins. Alternative production schemes are urgently needed for proteins that remain challenging to express and purify with conventional techniques. We have found that hydrophobin fusions targeted to endoplasmic reticulum (ER) can enhance the expression of target proteins simultaneously providing means for straightforward purification. Here we show that hydrophobin fusion technology induces formation of large protein bodies in the filamentous fungus Trichoderma reesei. The fusion protein remained soluble in the ER-derived protein bodies. A simple and scalable aqueous two-phase system was demonstrated to purify the hydrophobin fusion protein GFP-HFBI from the complex intracellular extracts with a recovery of up to 62%.

U2 - 10.1016/j.nbt.2011.09.006

DO - 10.1016/j.nbt.2011.09.006

M3 - Article

VL - 30

SP - 262

EP - 268

JO - New Biotechnology

JF - New Biotechnology

SN - 1871-6784

IS - 2

ER -