Online SFE-SFC-MS/MS colony screening: A high-throughput approach for optimizing (-)-limonene production

Area of Science:

• Biotechnology
• Analytical Chemistry
• Microbiology

Background:

• Traditional metabolite analysis from microbial cultures is time-intensive.
• Current methods limit the throughput for identifying bioproducers.
• Pretreatment steps in conventional analysis are laborious.

Purpose of the Study:

• To introduce a novel, high-throughput method for direct microbial colony analysis.
• To evaluate the efficacy of supercritical fluid extraction-supercritical fluid chromatography-triple quadrupole mass spectrometry (SFE-SFC-MS/MS) for this purpose.
• To demonstrate the application in screening (-)-limonene bioproducers.

Main Methods:

• Development and application of an online SFE-SFC-MS/MS system.
• Direct analysis of microbial colonies without prior metabolite extraction.
• Automated metabolite extraction integrated with chromatographic separation and mass spectrometry detection.

Main Results:

• The SFE-SFC-MS/MS system enables direct screening of microbial colonies.
• High-throughput analysis is achieved through automated, pretreatment-free metabolite extraction.
• Faster and more convenient screening of highly productive (-)-limonene bioproducer strains was demonstrated compared to conventional methods.

Conclusions:

• SFE-SFC-MS/MS is a pioneering method for direct microbial colony screening.
• This technique significantly enhances throughput and efficiency in identifying bioproducers.
• The system offers a streamlined approach for strain improvement and bioprocess development.