合成生物学 ›› 2023, Vol. 4 ›› Issue (5): 1000-1019.DOI: 10.12211/2096-8280.2023-031

• 特约评述 • 上一篇    下一篇

基于高通量液相色谱质谱技术的菌株筛选与关键分子定量分析研究进展

吴玉洁1,2,3, 刘欣欣1, 刘健慧1, 杨开广1, 随志刚1, 张丽华1, 张玉奎1   

  1. 1.中国科学院大连化学物理研究所,中国科学院分离分析化学重点实验室,辽宁 大连 116023
    2.大连理工大学化学学院,辽宁 大连 116024
    3.中国科学院大学,北京 100049
  • 收稿日期:2023-04-17 修回日期:2023-06-26 出版日期:2023-10-31 发布日期:2023-11-15
  • 通讯作者: 杨开广,随志刚
  • 作者简介:吴玉洁(1998—),女,博士研究生。研究方向为面向合成生物学的蛋白质组学分析方法。E-mail:yujiewu@dicp.ac.cn
    杨开广(1981—),男,研究员,博士生导师。研究方向为蛋白质组学新技术新方法。E-mail:yangkaiguang@dicp.ac.cn
    随志刚(1979—),男,副研究员,硕士生导师。研究方向为基于质谱的蛋白质及代谢物高通量分析方法开发。E-mail:zhigangsui@dicp.ac.cn
  • 基金资助:
    国家自然科学基金(22104138);中国科学院“青年创新促进会”优秀会员基金(Y2021058)

Research progress of strain screening and quantitative analysis of key molecules based on high-throughput liquid chromatography and mass spectrometry

Yujie WU1,2,3, Xinxin LIU1, Jianhui LIU1, Kaiguang Yang1, Zhigang SUI1, Lihua ZHANG1, Yukui ZHANG1   

  1. 1.Key Laboratory of Separation Science for Analytical Chemistry,Dalian Institute of Chemical Physics,Chinese Academy of Sciences,Dalian 116023,Liaoning,China
    2.School of Chemistry,Dalian University of Technology,Dalian 116024,Liaoning,China
    3.University of Chinese Academy of Sciences,Beijing 100049,China
  • Received:2023-04-17 Revised:2023-06-26 Online:2023-10-31 Published:2023-11-15
  • Contact: Kaiguang Yang, Zhigang SUI

摘要:

合成生物学是21世纪新兴的交叉学科,已经为医药、化工、能源、食品和农业等领域带来了显著的改变。微生物细胞工厂(microbial cell factory,MCF)的构建和应用是合成生物学的重要研究内容,正迅速向实用化、产业化的方向发展。得益于基因编辑技术的进步,MCF菌株文库构建的能力大幅提升,亦产生了数量庞大的待测样本,亟需发展高通量自动化的分析检测方法与之匹配。此外,针对关键代谢酶或代谢物的高通量检测技术,也对MCF改造乃至工业化生产有重要的指导意义。液相色谱质谱技术是生物分子检测分析的重要手段,在蛋白质、代谢物定量分析方面占据重要地位。因此本文总结回顾了近年来高通量液相色谱质谱技术在微生物菌株筛选及关键分子定量分析方面的研究进展,从样品制备、色谱分离、质谱分析及数据处理等层面展开阐述,并对这一领域与自动化平台结合的前景及如何深度融合的挑战进行简要概括,为推动基于合成生物学生物智造领域的快速发展提供指导。

关键词: 液相色谱, 质谱, 高通量筛选, 蛋白质组学, 代谢组学, 合成生物学, 微生物细胞工厂

Abstract:

Synthetic biology is an emerging interdisciplinary research area and has brought significant changes to fields such as medicine, chemical engineering, energy, food and agriculture. The construction and application of microbial cell factories (MCFs) is an important task of synthetic biology, which will lead the way to a sustainable industrial-scale manufacturing sector. With the progress of genome editing technology, the construction strategy of MCFs has evolved from random mutation to customized transformation at the whole genome level. Biologists can obtain 103~107 strain mutant libraries in a short time, creating an urgent need for the development of high-throughput screening methods. In addition, the demand for precise quantitative analysis of key molecules in metabolic pathways, such as metabolic enzymes and metabolites, is increasingly prominent in the process of strain transformation, evolution, and fermentation monitoring. Liquid chromatography (LC) offers excellent separation capability, allowing efficient separation of target molecules, while mass spectrometry (MS) provides strong detection ability due to its high specificity and sensitivity. Therefore, LC and MS techniques have been widely applied in the field of life sciences, especially for the qualitative and quantitative analyses of proteins and metabolites. With the development of high-throughput LC and MS technologies, these methods have become powerful tools for screening strains and quantitative research of key molecules in synthetic biology. Therefore, this work reviews the research progresses of LC and/or MS based strain screening and quantification of key molecules, focusing on the following aspects: sample preparation, chromatographic separation, mass spectrometric analysis and data processing. Finally, we briefly summarize the future prospects and challenges in this field. It is expected that with the continuous progress of LC and MS, the deep integration with automatic devices and data processing platforms, the advantages of LC and MS in high-throughput screening of synthetic biology strains and quantitative analysis of key molecules will become increasingly prominent, which will certainly promote the rapid development of biological intelligence based on synthetic biology.

Key words: liquid chromatography, mass spectrometry, high-throughput screening, proteomics, metabolomics, synthetic biology, microbial cell factory

中图分类号: