Soluble Expression and Efficient Purification of Recombinant Class I Hydrophobin DewA
Hydrophobins are small proteins (<20 kDa) with an amphipathic tertiary structure that are secreted by various filamentous fungi. Their amphipathic properties provide surfactant-like activity, leading to the formation of robust amphipathic layers at hydrophilic–hydrophobic interfaces, which make t...
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MDPI AG
2021-07-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/22/15/7843 |
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| author | Sang-Oh Ahn Ho-Dong Lim Sung-Hwan You Dae-Eun Cheong Geun-Joong Kim |
| author_facet | Sang-Oh Ahn Ho-Dong Lim Sung-Hwan You Dae-Eun Cheong Geun-Joong Kim |
| author_sort | Sang-Oh Ahn |
| collection | DOAJ |
| description | Hydrophobins are small proteins (<20 kDa) with an amphipathic tertiary structure that are secreted by various filamentous fungi. Their amphipathic properties provide surfactant-like activity, leading to the formation of robust amphipathic layers at hydrophilic–hydrophobic interfaces, which make them useful for a wide variety of industrial fields spanning protein immobilization to surface functionalization. However, the industrial use of recombinant hydrophobins has been hampered due to low yield from inclusion bodies owing to the complicated process, including an auxiliary refolding step. Herein, we report the soluble expression of a recombinant class I hydrophobin DewA originating from <i>Aspergillus nidulans,</i> and its efficient purification from recombinant <i>Escherichia coli</i>. Soluble expression of the recombinant hydrophobin DewA was achieved by a tagging strategy using a systematically designed expression tag (ramp tag) that was fused to the N-terminus of DewA lacking the innate signal sequence. Highly expressed recombinant hydrophobin DewA in a soluble form was efficiently purified by a modified aqueous two-phase separation technique using isopropyl alcohol. Our approach for expression and purification of the recombinant hydrophobin DewA in <i>E. coli</i> shed light on the industrial production of hydrophobins from prokaryotic hosts. |
| first_indexed | 2024-03-10T09:14:53Z |
| format | Article |
| id | doaj.art-545323bdfa8d4698a9bc14d13f0ac57c |
| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-10T09:14:53Z |
| publishDate | 2021-07-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-545323bdfa8d4698a9bc14d13f0ac57c2023-11-22T05:39:17ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672021-07-012215784310.3390/ijms22157843Soluble Expression and Efficient Purification of Recombinant Class I Hydrophobin DewASang-Oh Ahn0Ho-Dong Lim1Sung-Hwan You2Dae-Eun Cheong3Geun-Joong Kim4Department of Biological Sciences, College of Natural Sciences, Chonnam National University, Yongbong-ro, Buk-gu, Gwangju 61186, KoreaCenter for Industrialization of Agricultural and Livestock Microorganisms, 241 Cheomdangwahak-ro, Jeongeup-si 56212, Jeollabuk-do, KoreaBiomedical Research Center, Chonnam National University, Convergence Science Building (M2), Suite 301-1 264, Seoyang-ro, Hwasun-eup, Hwasun-gun 58128, Jeollanam-do, KoreaDepartment of Biological Sciences, College of Natural Sciences, Chonnam National University, Yongbong-ro, Buk-gu, Gwangju 61186, KoreaDepartment of Biological Sciences, College of Natural Sciences, Chonnam National University, Yongbong-ro, Buk-gu, Gwangju 61186, KoreaHydrophobins are small proteins (<20 kDa) with an amphipathic tertiary structure that are secreted by various filamentous fungi. Their amphipathic properties provide surfactant-like activity, leading to the formation of robust amphipathic layers at hydrophilic–hydrophobic interfaces, which make them useful for a wide variety of industrial fields spanning protein immobilization to surface functionalization. However, the industrial use of recombinant hydrophobins has been hampered due to low yield from inclusion bodies owing to the complicated process, including an auxiliary refolding step. Herein, we report the soluble expression of a recombinant class I hydrophobin DewA originating from <i>Aspergillus nidulans,</i> and its efficient purification from recombinant <i>Escherichia coli</i>. Soluble expression of the recombinant hydrophobin DewA was achieved by a tagging strategy using a systematically designed expression tag (ramp tag) that was fused to the N-terminus of DewA lacking the innate signal sequence. Highly expressed recombinant hydrophobin DewA in a soluble form was efficiently purified by a modified aqueous two-phase separation technique using isopropyl alcohol. Our approach for expression and purification of the recombinant hydrophobin DewA in <i>E. coli</i> shed light on the industrial production of hydrophobins from prokaryotic hosts.https://www.mdpi.com/1422-0067/22/15/7843ramp tag (RT)soluble expressionrecombinant hydrophobin DewAaqueous two-phase separation (ATPS)isopropyl alcohol (IPA) |
| spellingShingle | Sang-Oh Ahn Ho-Dong Lim Sung-Hwan You Dae-Eun Cheong Geun-Joong Kim Soluble Expression and Efficient Purification of Recombinant Class I Hydrophobin DewA International Journal of Molecular Sciences ramp tag (RT) soluble expression recombinant hydrophobin DewA aqueous two-phase separation (ATPS) isopropyl alcohol (IPA) |
| title | Soluble Expression and Efficient Purification of Recombinant Class I Hydrophobin DewA |
| title_full | Soluble Expression and Efficient Purification of Recombinant Class I Hydrophobin DewA |
| title_fullStr | Soluble Expression and Efficient Purification of Recombinant Class I Hydrophobin DewA |
| title_full_unstemmed | Soluble Expression and Efficient Purification of Recombinant Class I Hydrophobin DewA |
| title_short | Soluble Expression and Efficient Purification of Recombinant Class I Hydrophobin DewA |
| title_sort | soluble expression and efficient purification of recombinant class i hydrophobin dewa |
| topic | ramp tag (RT) soluble expression recombinant hydrophobin DewA aqueous two-phase separation (ATPS) isopropyl alcohol (IPA) |
| url | https://www.mdpi.com/1422-0067/22/15/7843 |
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