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.

疏水蛋白是真菌分泌的低分子量蛋白，具有极强的表面活性，能够自组装成更大的结构。由于疏水蛋白具有非凡的生化特性，因此它们在药物乳化和表面改性等商业应用中成为有吸引力的目标。当在大肠杆菌中产生时，疏水蛋白通常不溶，需要重新折叠。在这项工作中，我们将SHuffle T7 Express大肠杆菌与谷胱甘肽氧化还原缓冲液结合使用，以产生并重折叠源自Phanerochaete carnosa（PC1），Wallemia ichthyophaga（WI1），Serpula lacrymans（SL1）和Schizophyllum communeune的四种不同的IB类疏水蛋白。（SC16）。这些纯化的疏水蛋白的正确重折叠和功能已通过核磁共振波谱和硫黄素T分析得以证实。这些结果表明，可以始终如一地从大肠杆菌生产和纯化IB类疏水蛋白，以帮助将来需要高纯度疏水蛋白的结构和生化研究。