Pei Xiaolin, Wang Jiapao, Zheng Haoteng, Xiao Qinjie, Wang Anming, Su Weike
Collaborative Innovation Center of Yangtze River Delta Region Green Pharmaceuticals, Zhejiang University of Technology, Hangzhou, China; College of Material, Chemistry and Chemical Engineering, Hangzhou Normal University, Hangzhou, China.
College of Material, Chemistry and Chemical Engineering, Hangzhou Normal University, Hangzhou, China.
Enzyme Microb Technol. 2022 Jan;153:109915. doi: 10.1016/j.enzmictec.2021.109915. Epub 2021 Sep 9.
The catalytically-active inclusion bodies (CatIBs) represent a promising strategy for immobilizing enzyme without additional carriers and chemicals, which has aroused great attention in academic and industrial communities. In this work, we discovered two natural parallel right-handed coiled-coil tetramer peptides from PDB database by a structural mining strategy. The two self-assembling peptides, NSPdoT from rotavirus and HVdoT from human Vasodilator-stimulated phosphoprotein, efficiently induced the CatIBs formation of a (R)-Hydroxynitrile lyase from Arabidopsis thaliana (AtHNL) in Escherichia coli cells. This is convenient to simultaneously purify and immobilize the target proteins as biocatalysts. As expected, HVdoT-AtHNL and NSPdoT-AtHNL possessed drastically increased tolerance toward lower pH values, which will be very critical to synthesize cyanohydrins under acidic condition for suppressing the non-enzymatic side reaction. In addition. AtHNL-CatIBs are produced at high yield in host cells as bioactive microparticles, which exhibited high thermal and pH stabilities. Therefore, the CatIBs method represent a promising application for the immobilization of enzymes in the biocatalysis field.
具有催化活性的包涵体（CatIBs）是一种无需额外载体和化学物质即可固定化酶的有前景的策略，已在学术界和工业界引起了极大关注。在这项工作中，我们通过结构挖掘策略从蛋白质数据银行（PDB）数据库中发现了两种天然的平行右手卷曲螺旋四聚体肽。这两种自组装肽，即来自轮状病毒的NSPdoT和来自人血管舒张刺激磷蛋白的HVdoT，在大肠杆菌细胞中有效地诱导了来自拟南芥的（R）-羟基腈裂解酶（AtHNL）形成具有催化活性的包涵体。这便于同时纯化和固定化作为生物催化剂的目标蛋白。正如预期的那样，HVdoT-AtHNL和NSPdoT-AtHNL对较低pH值的耐受性大幅提高，这对于在酸性条件下合成氰醇以抑制非酶促副反应至关重要。此外，AtHNL-CatIBs在宿主细胞中作为生物活性微粒高产产生，表现出高热稳定性和pH稳定性。因此，具有催化活性的包涵体方法在生物催化领域的酶固定化方面具有广阔的应用前景。
