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Fungal Hydrophobin RolA Enhanced PETase Hydrolysis of Polyethylene Terephthalate.
Applied Biochemistry and Biotechnology ( IF 3.1 ) Pub Date : 2020-06-06 , DOI: 10.1007/s12010-020-03358-y
Nathania Puspitasari , Shen-Long Tsai , Cheng-Kang Lee

Polyethylene terephthalate (PET) becomes one of the most well-known polyesters and is widely used as packaging material. Recently, polyethylene terephthalate hydrolase (PETase) has emerged as a potential biocatalyst demonstrating the ability to degrade polyethylene terephthalate (PET). We showed that the rate of PETase hydrolysis could be significantly increased in the presence of hydrophobin RolA. Hydrophobins represent a class of small fungal protein that has a high surface-active substance and can spontaneously self-assemble at hydrophilic-hydrophobic interfaces. In this work, a class I hydrophobin named RolA was extracted from the mycelium pellet collected from a fermentation culture of Aspergillus oryzae. The SDS-PAGE analysis of the isolated RolA showed the presence of 11 kDa polypeptide. Recombinant PETase from Ideonella sakaiensis was also successfully expressed in Escherichia coli as a soluble protein with molecular weight approximately 30 kDa. The hydrophobin RolA could enhance the PET hydrolysis in the presence of the recombinant PETase. The hydrolysis of PET bottle by RolA-PETase achieved the highest weight loss of 26% in 4 days. It is speculated that the wetting effect of RolA acts on PET surface converts PET to become hydrophilic that leads PETase easier to contact and attack the surface.

Graphical Abstract



中文翻译:

真菌疏水蛋白 RolA 增强聚对苯二甲酸乙二醇酯的 PETase 水解。

聚对苯二甲酸乙二醇酯 (PET) 成为最知名的聚酯之一,被广泛用作包装材料。最近,聚对苯二甲酸乙二醇酯水解酶 (PETase) 已成为一种潜在的生物催化剂,展示了降解聚对苯二甲酸乙二醇酯 (PET) 的能力。我们表明,在疏水蛋白 RolA 的存在下,PETase 水解的速率可以显着增加。疏水蛋白是一类具有高表面活性物质的小型真菌蛋白,可以在亲水-疏水界面自发自组装。在这项工作中,从米曲霉发酵培养物中收集的菌丝体颗粒中提取了一种名为 RolA 的 I 类疏水蛋白. 分离的 RolA 的 SDS-PAGE 分析表明存在 11 kDa 多肽。来自Ideonella sakaiensis 的重组 PETase也在大肠杆菌中成功表达为分子量约为 30 kDa 的可溶性蛋白质。在重组 PETase 存在的情况下,疏水蛋白 RolA 可以增强 PET 水解。RolA-PETase 对 PET 瓶的水解达到了 26% 的最高重量损失在 4 天内。据推测,RolA 的润湿作用作用于 PET 表面,使 PET 转变为亲水性,使 PETase 更容易接触和攻击表面。

图形概要

更新日期:2020-06-06
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