Yang Jun, Deng Binyang, Liao Pingan, Lin Siyu, Zheng Liqi, Yang Xing, Wang Fei, Zhai Chao, Ma Lixin
State Key Laboratory of Biocatalysis and Enzyme Engineering, Hubei Key Laboratory of Industrial Biotechnology, School of Life Sciences, Hubei University, Wuhan 430062, China.
Biomolecules. 2025 Jun 18;15(6):888. doi: 10.3390/biom15060888.
Previous reports indicated that self-assembling amphipathic peptide S1v1 (AEAEAHAH) significantly enhances the soluble expression, thermostability, and activity of the target proteins when fused to them. In order to obtain high-efficiency enzymes for the large-scale degradation of polyethylene terephthalate (PET), this multifunctional peptide was fused to the N- and C-terminus of FAST-PETase, a variant of PETase (PETase), with a PT-linker (TTVTTPQTS) harbored between the target protein and the multifunctional peptide. Consistent with previous reports, S1v1 increased the solubility of FAST-PETase slightly. Moreover, it increased the activity of FAST-PETase dramatically. The amount of terephthalic acid (TPA) and mono(2-hydroxyethyl) terephthalic acid (MHET) released from PET substrate after 24 h of digestion at 50°C by fusion enzymes bearing N- and C-terminal S1v1 tag was approximately 2.9- and 4.6-fold that of FAST-PETase, respectively. Furthermore, the optimal temperature and thermostability of the fusion proteins increased in comparison with FAST-PETase. The present study provides a novel strategy to improve the depolymerization efficiency of FAST-PETase.
先前的报道表明，自组装两亲性肽S1v1（AEAEAHAH）与目标蛋白融合时，能显著提高其可溶性表达、热稳定性及活性。为了获得用于大规模降解聚对苯二甲酸乙二酯（PET）的高效酶，该多功能肽与PET酶（PETase）的变体FAST-PETase的N端和C端融合，并在目标蛋白与多功能肽之间含有一个PT连接子（TTVTTPQTS）。与先前报道一致，S1v1略微提高了FAST-PETase的溶解度。此外，它还显著提高了FAST-PETase的活性。在50℃下消化24小时后，带有N端和C端S1v1标签的融合酶从PET底物中释放的对苯二甲酸（TPA）和单（2-羟乙基）对苯二甲酸（MHET）的量分别约为FAST-PETase的2.9倍和4.6倍。此外，与FAST-PETase相比，融合蛋白的最佳温度和热稳定性有所提高。本研究为提高FAST-PETase的解聚效率提供了一种新策略。
