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Recently, the research group of Si Tong, Institute of Synthetic Biology, Shenzhen Institutes of Advanced Technology, Chinese Academy of Sciences, published an invited review article online in the internationally renowned academic journal Current Opinion in Biotechnology: Towards one sample per second for mass spectrometric screening of engineered microbial strains
.
The article focuses on the high-throughput mass spectrometry screening method of engineered strains (1 sec/sample), and reviews the microbial culture system, automated sample preparation, high-throughput mass spectrometry instruments, strain development and application, etc.
, and discusses the key challenges and future trends.
Outlook
.
Researcher Si Tong and assistant researcher Chen Yongcan are the co-corresponding authors of this article, and doctoral student Fu Lihao and research assistant Guo Erpeng are the co-first authors
.
Microbial cell factories hold promise for sustainable production of products such as fuels, chemicals, and materials, but require systematic engineering of strains to meet industrial production needs
.
At present, the success rate of rational design of cell factories is low, and strain development needs to rely on high-throughput library construction and screening
.
Although mass spectrometry has the advantages of high sensitivity, high specificity, high accuracy, and no labeling, traditional chromatography-mass spectrometry methods have low throughput and long cycle times, which cannot meet the needs of rapid strain screening
.
The article proposes that omitting the chromatographic separation step is essential to increase the throughput of mass spectrometry
.
Microbial samples can directly enter the mass spectrometer by means of autosampler, laser/ion/gas beam, ultrasound, microfluidics, etc.
; and sample preparation is a key step in connecting various culture systems and mass spectrometry platforms (Figure 1)
.
Figure 1.
High-throughput mass spectrometry analysis schemes for different cultures.
For common microbial culture systems such as agar plate colonies, microplates, and microfluidic droplets, the article discusses the latest progress in this field from sample preparation, mass spectrometry equipment, and data analysis.
emphasised
.
The authors select a selection of commercial or application-proven mass spectrometry methods for introduction and comparison, and summarize screening throughput and application examples (Table 1)
.
The application of high-throughput mass spectrometry screening in the development of rhamnolipid, medium-chain fatty acid, lysine, sweet protein and other production strains is illustrated
.
Table 1 High-throughput mass spectrometry analysis platform for microbial screening The article finally analyzes the unique challenges of microbial culture systems to high-throughput mass spectrometry screening methods, and points out that factors such as screening throughput, chemical coverage, and quantitative accuracy should be weighed in method development.
, select and develop the most suitable culture system, sample preparation method and mass spectrometry equipment for a specific application
.
Brief introduction of PI and research group: Si Tong, researcher and doctoral supervisor of the Institute of Synthetic Biology, Shenzhen Academy of Advanced Sciences, Chinese Academy of Sciences
.
The chief scientist of the synthetic biology project of the National Key R&D Program, the national high-level talent (youth), and the chief technologist of Shenzhen's synthetic biology research major scientific and technological infrastructure
.
The research group's direction is automated synthetic biotechnology, including machine learning-guided protein engineering, high-throughput mass spectrometry screening, etc.
, for the development of microbial cell factories to research and produce important molecules such as fuels, chemicals, and drugs.
The previous results are in Nature Communications, JACS, He has published more than 40 papers in internationally renowned academic journals such as Angew Chemie and Metabolic Engineering, and has been cited more than 2,200 times by Google Scholar
.
The research group has long been committed to the development of high-throughput mass spectrometry instruments and new methods for synthetic biology research.
We are now recruiting postdoctoral fellows, engineers and visiting students in the fields of mass spectrometry and instrument analysis.
Interested applicants please email your resume to tong.
si@ siat.
ac.
cn
.
Laboratory homepage: http://isynbio.
siat.
ac.
cn/sitonglab/ Paper link: https://authors.
elsevier.
com/a/1e%7EAA3PtAV1NZH Open for reprinting, welcome to forward to Moments and WeChat groups