摘要
在单分子工具和技术的迅速发展过程中,单分子荧光显微镜技术脱颖而出,可以高特异性地监测荧光标记的生物分子的时空行为,成为研究生物系统的有力工具。然而,单分子成像和追踪技术在工业微生物研究及酶工程领域中的应用仍处于起步阶段。随着生物工程技术、合成生物学和代谢工程等科学技术的迅猛发展,工业微生物的应用日趋广泛。但是,菌株往往难以在整个培养期间保持最大的生产能力,导致在经济效益上不能满足产业化的需求,成为制约微生物细胞工厂发展的一个瓶颈,对酶学研究提出新的挑战。单分子成像和追踪技术具有直接、准确、实时等监测优点,可以直接在活体生物上进行研究,成功地实现了对单分子酶催化过程的实时监控,发现了多种酶的新单分子行为及反应机制。这些技术有助于各种新型酶、新特性酶、新作用方式酶的研发。本文简述单分子成像和追踪技术的主要特点(以荧光显微镜为主),总结近年来单分子成像和追踪技术在微生物研究、酶学研究中的应用,及其在工业微生物研究中的应用现状和面临的挑战。这些技术的应用必将促进纳米生物学的发展,推动酶工程改造,提高细胞工厂的生产能力。
Single molecule tools and technologies have being developed rapidly,and among them single molecule fluorescence microscope technology stands out,which can monitor the spatiotemporal behavior of fluorescent labeled bio-molecules with high specificity,and has become a powerful tool for studying biological systems.However,the application of single molecule imaging and tracking technology is still in its infancy in industrial microbial research and enzyme engineering.With the rapid development of bioengineering technology,synthetic biology and metabolic engineering,industrial microorganisms are widely used.However,it is often difficult for strains to maintain the maximum production capacity during the whole cultivation period,which leads to the economic benefits that cannot meet the needs of industrialization,and becomes a bottleneck problem restricting the development of microbial cell factory.Thus,new challenges have been posed to the enzymatic research in the field of enzyme engineering.The technology of single molecule imaging and tracking has the advantages of direct,accurate and real-time monitoring,which can be directly studied on living organisms.It has successfully realized the real-time monitoring of the catalytic process of single molecular enzymes,and found new single molecular behaviors and reaction mechanisms of various enzymes.These technologies contribute to the development of various new enzymes,new characteristic enzymes and new modes of action enzymes.The main characteristics of single molecule imaging and tracking technologies with fluorescence microscopes are briefly described in this article.The application of single molecule imaging technology in microbial research and enzymology research in the field of enzyme engineering in recent years,as well as its application status and challenges in industrial microbial research are summarized.The application of these technologies will promote the development of nanobiology,promote the engineering transformation of enzymes and improve the production capacity of cell factories.
作者
严少敏
吴光
YAN Shaomin;WU Guang(National Engineering Research Center for Non-Food Biorefinery,State Key Laboratory of Non-Food Biomass and Enzyme Technology,Guangxi Biomass Engineering Technology Research Center,Guangxi Key Laboratory of Biorefinery,Guangxi Academy of Sciences,Nanning,Guangxi,530007,China)
出处
《广西科学》
CAS
北大核心
2021年第5期429-439,共11页
Guangxi Sciences
基金
广西重点研发计划项目(桂科AB17195034)
广西科学院科研发展基金项目(2021YFJ1203)资助。
关键词
单分子成像
单分子追踪
工业微生物
酶工程
荧光显微镜
single molecule imaging
single molecule tracking
industrial microorganism
enzyme engineering
fluorescence microscope