Andreu Cecilia, Gómez-Peinado Javier, Winandy Lex, Fischer Reinhard, Del Olmo Marcel Li
Departament de Química Orgànica, Facultat de Farmàcia, Universitat de València, Burjassot, València, Spain.
Departament de Bioquímica i Biologia Molecular, Facultat de Ciències Biològiques, Universitat de València, Burjassot, València, Spain.
Appl Microbiol Biotechnol. 2021 Feb;105(4):1505-1518. doi: 10.1007/s00253-021-11090-8. Epub 2021 Jan 23.
Hydrophobins are relatively small proteins produced naturally by filamentous fungi with interesting biotechnological and biomedical applications given their self-assembly capacity, efficient adherence to natural and artificial surfaces, and to introduce modifications on the hydrophobicity/hydrophilicity of surfaces. In this work we demonstrate the efficient expression on the S. cerevisiae cell surface of class II HFBI of Trichoderma reesei and class I DewA of Aspergillus nidulans, a hydrophobin not previously exposed, using the Yeast Surface Display a-agglutinin (Aga1-Aga2) system. We show that the resulting modifications affect surface properties, and also yeast cells' resistance to several adverse conditions. The fact that viability of the engineered strains increases under heat and osmotic stress is particularly interesting. Besides, improved biocatalytic activity toward the reduction of ketone 1-phenoxypropan-2-one takes place in the reactions carried out at both 30 °C and 40 °C, within a concentration range between 0.65 and 2.5 mg/mL. These results suggest interesting potential applications for hydrophobin-exposing yeasts. KEY POINTS : • Class I hydrophobin DewA can be efficiently exposed on S. cerevisiae cell surfaces. • Yeast exposure of HFBI and DewA increases osmotic and heat resistance. • Engineered strains show modified biocatalytic behavior.
疏水蛋白是丝状真菌天然产生的相对较小的蛋白质,鉴于其自组装能力、对天然和人工表面的有效粘附以及对表面疏水性/亲水性的修饰能力,具有有趣的生物技术和生物医学应用。在这项工作中,我们利用酵母表面展示α-凝集素(Aga1-Aga2)系统,证明了里氏木霉II类HFBI和烟曲霉I类DewA(一种以前未被展示的疏水蛋白)在酿酒酵母细胞表面的高效表达。我们表明,由此产生的修饰会影响表面性质,以及酵母细胞对几种不利条件的抗性。工程菌株在热应激和渗透应激下活力增加这一事实尤其有趣。此外,在30℃和40℃下进行的反应中,在0.65至2.5mg/mL的浓度范围内,对1-苯氧基丙酮-2-酮还原的生物催化活性有所提高。这些结果表明疏水蛋白展示酵母具有有趣的潜在应用。关键点:• I类疏水蛋白DewA可以在酿酒酵母细胞表面高效展示。• HFBI和DewA在酵母表面的展示增加了渗透压和耐热性。• 工程菌株表现出修饰后的生物催化行为。