聚对苯二甲酸乙二醇酯(PET)水解酶的高通量筛选:进展与挑战  

High-throughput screening assays for PET hydrolases:progress and challenges

作　　者：韦韧 WEI Ren(Institute of Biochemistry,University of Greifswald,Greifswald 17489,Germany)

机构地区：[1]格莱夫斯瓦尔德大学生物化学研究所,德国格莱夫斯瓦尔德17489

出　　处：《生物加工过程》2022年第4期353-364,共12页Chinese Journal of Bioprocess Engineering

基　　金：欧盟地平线2020计划科研创新行动(870294);国家自然科学基金(31961133017)。

摘　　要：聚对苯二甲酸乙二醇酯(PET)是世界上最常见且用途广泛的热塑性聚酯塑料。由于PET具有较高的化学惰性和环境稳定性,使用后处理不当的PET废弃物在全球范围内持续积累,造成了环境污染和生态危机。近年来,在塑料循环经济的框架内,开发新型PET废弃物回收和升级改造技术成为全球关注的发展方向之一。自然界中丰富的微生物资源为发掘、改造酶生物催化剂用于降解及回收PET塑料废弃物提供了可能性。从2005年第一个PET水解酶被报道至今十几年的时间里,这个领域的研究包括机制分析、生化表征和蛋白质工程等,一直局限于几种基准酶。由于缺少有效的专门针对PET水解活性的高通量筛选方法,人们无法更全面地检索各种微生物资源,也无法从大规模的突变体文库中迅速发现活性最高的酶。近年来,人们对常见表征PET水解酶的分析方法进行了各种改良并尝试将它们应用于高通量筛选。本篇综述概括了这个方法学领域的最新进展,并以PET纳米颗粒、紫外/可见光分光光度法、荧光分光光度法以及生物传感器等策略为例子,探讨了高通量PET水解酶筛选的可行性、目前的技术困境以及未来可能取得突破的一些方案。Polyethylene terephthalate(PET)is the most commonly used thermoplastic polyester on a global scale.Due to its high resistance to various physical and chemical factors,PET waste can persist for a very long period in natural ecosystems where it can cause serious environmental pollution and ecological disasters.Within the framework of the circular plastic economy,the development of innovative PET waste recycling and upcycling technologies has become a global trend in recent years.The abundance of microorganisms in nature allows for the isolation of novel enzymes capable of degrading and recycling PET plastic waste.Since the first PET hydrolase was discovered in 2005,this research field,which includes mechanistic investigation,biochemical characterization,and protein engineering,has been limited to a small number of benchmark enzymes.Due to the lack of efficient high-throughput screening methods for PET hydrolysis activity,a more thorough exploration of varied microbial resources and the rapid identification of the most active enzymes from huge mutant libraries has been hampered.Several attempts have been made to adapt traditional analytical methods for characterizing PET hydrolases to high-throughput screening scenarios.This review focuses on current findings in this field.Using recent developments such as PET nanoparticles generation,UV/visible spectrophotometry,fluorescence spectrophotometry,and biosensors as examples,this review explores the feasibility and current technical challenges of high-throughput PET hydrolase screening assays,and offers options for potential breakthroughs.

关 键 词：高通量筛选 PET水解酶 纳米颗粒 对苯二甲酸酯 比色法 荧光法 生物传感器 循环经济 

分 类 号：Q556[生物学—生物化学] X705[环境科学与工程—环境工程] X172