Department of Biological Sciences, College of Natural Sciences, Chonnam National University, Yongbong-ro, Buk-gu, Gwangju 61186, Korea.
Center for Industrialization of Agricultural and Livestock Microorganisms, 241 Cheomdangwahak-ro, Jeongeup-si 56212, Jeollabuk-do, Korea.
Int J Mol Sci. 2021 Jul 22;22(15):7843. doi: 10.3390/ijms22157843.
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 and its efficient purification from recombinant . 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 shed light on the industrial production of hydrophobins from prokaryotic hosts.
水蛋白是由各种丝状真菌分泌的小蛋白（<20 kDa），具有两亲性三级结构。它们的两亲性质提供了表面活性剂样的活性，导致在亲水-疏水界面形成坚固的两亲性层，这使得它们在跨越蛋白质固定化到表面功能化的各种工业领域都非常有用。然而，由于包含体的产量低（由于辅助重折叠步骤，包括复杂的过程），重组水蛋白的工业应用受到了阻碍。在此，我们报道了一种来源于 的重组 I 类水蛋白 DewA 的可溶表达及其从重组 中的有效纯化。通过使用系统设计的表达标签（ramp tag）融合到缺乏天然信号序列的 DewA 的 N 端的标记策略实现了重组水蛋白 DewA 的可溶表达。使用异丙醇的改良双水相分离技术可高效地从可溶性形式中纯化高表达的重组水蛋白 DewA。我们在 中表达和纯化重组水蛋白 DewA 的方法为从原核宿主生产水蛋白提供了思路。
