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

WU Yujie; LIU Xinxin; LIU Jianhui; Yang Kaiguang; SUI Zhigang; ZHANG Lihua; ZHANG Yukui

doi:10.12211/2096-8280.2023-031

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

Invited Review | 更新时间：2025-03-20

- Research progress of strain screening and quantitative analysis of key molecules based on high-throughput liquid chromatography and mass spectrometry
- Synthetic Biology Journal Vol. 4, Issue 5, Pages: 1000-1019(2023)
- 作者机构：
  
  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
- 作者简介：
- 基金信息：
- DOI：10.12211/2096-8280.2023-031
  CLC： O657.63
- Received：17 April 2023，
  
  Revised：2023-06-26，
  
  Published：31 October 2023
- 稿件说明：
移动端阅览
吴玉洁，刘欣欣，刘健慧，杨开广，随志刚，张丽华，张玉奎. 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［J］. 合成生物学， 2023， 4（5）： 1000-1019

WU Yujie， LIU Xinxin， LIU Jianhui， Yang Kaiguang， SUI Zhigang， ZHANG Lihua， ZHANG Yukui. Research progress of strain screening and quantitative analysis of key molecules based on high-throughput liquid chromatography and mass spectrometry［J］. Synthetic Biology Journal， 2023， 4（5）： 1000-1019
吴玉洁，刘欣欣，刘健慧，杨开广，随志刚，张丽华，张玉奎. 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［J］. 合成生物学， 2023， 4（5）： 1000-1019 DOI： 10.12211/2096-8280.2023-031.

WU Yujie， LIU Xinxin， LIU Jianhui， Yang Kaiguang， SUI Zhigang， ZHANG Lihua， ZHANG Yukui. Research progress of strain screening and quantitative analysis of key molecules based on high-throughput liquid chromatography and mass spectrometry［J］. Synthetic Biology Journal， 2023， 4（5）： 1000-1019 DOI： 10.12211/2096-8280.2023-031.

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。

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 10

~10

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 mole

cules 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.

关键词

Keywords

references

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