通过信号肽增强子表达 IsPETase 并增强 PET 水解在大肠杆菌中的分泌表达。
Excretory expression of IsPETase in E. coli by an enhancer of signal peptides and enhanced PET hydrolysis.
机构信息
College of Biotechnology and Pharmaceutical Engineering, Nanjing Tech University, No. 30 Puzhu South Road, Nanjing 211816, China.
School of Pharmaceutical Sciences, Nanjing Tech University, No. 30 Puzhu South Road, Nanjing 211816, China.
出版信息
Int J Biol Macromol. 2021 Oct 1;188:568-575. doi: 10.1016/j.ijbiomac.2021.08.012. Epub 2021 Aug 8.
The PET hydrolase from Ideonella sakaiensis (IsPETase) is efficient for PET degradation, which provides a promising solution for environmental contamination by plastics. This study focuses on improving the excretion of IsPETase from E. coli by signal peptide (SP) engineering. A SP enhancer B1 (MERACVAV) was fused to the N-terminal of commonly-used SP (PelB, MalE, LamB, and OmpA) to mediate excretion of IsPETase. Strikingly, the modified SP B1OmpA, B1PelB, and B1MalE significantly increased the excretion of IsPETase, while IsPETase was basically expressed in periplasmic space without enhancer B1. The excretion efficiency of IsPETase mediated by B1PelB was improved by 62 folds compared to that of PelB. The hydrolysis of PET by crude IsPETase in culture solution was also enhanced. Furthermore, the amount of released MHET/TPA from PET by IsPETase was increased by 2.7 folds with pre-incubation of hydrophobin HFBII. Taken together, this work may provide a feasible strategy for the excretion and application of the IsPETase.
来自门氏菌属（Ideonella sakaiensis）的 PET 水解酶（IsPETase）在 PET 降解方面效率很高，为解决塑料对环境的污染提供了很有前景的解决方案。本研究重点通过信号肽（SP）工程来提高 IsPETase 从大肠杆菌中的分泌。一个 SP 增强子 B1（MERACVAV）被融合到常用 SP（PelB、MalE、LamB 和 OmpA）的 N 端，以介导 IsPETase 的分泌。引人注目的是，修饰后的 SP B1OmpA、B1PelB 和 B1MalE 显著提高了 IsPETase 的分泌，而没有增强子 B1 时，IsPETase 基本在周质空间表达。与 PelB 相比，B1PelB 介导的 IsPETase 分泌效率提高了 62 倍。在培养溶液中，粗制的 IsPETase 对 PET 的水解作用也得到了增强。此外，通过预先孵育疏水蛋白 HFBII，IsPETase 从 PET 中释放的 MHET/TPA 量增加了 2.7 倍。总之，这项工作可能为 IsPETase 的分泌和应用提供了一种可行的策略。
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