Rapid analysis of terpenes produced by fermentation using flow injection analysis coupled to APCI MS

Area of Science:

• Biotechnology
• Analytical Chemistry
• Synthetic Biology

Background:

• Terpenes are vital compounds with broad industrial applications.
• Enzyme- and microorganism-based synthesis offers selective and eco-friendly terpene production.
• Efficient screening of engineered strains is crucial for optimizing terpene yields.

Purpose of the Study:

• To develop a high-throughput, label-free screening method for terpenes.
• To enable rapid quantification and adaptation for various terpene analytes.
• To accelerate the engineering of microorganisms for enhanced terpene biosynthesis.

Main Methods:

• Developed a flow injection analysis (FIA) method.
• Utilized atmospheric pressure chemical ionization (APCI) coupled to a quadrupole time-of-flight (Q-TOF) mass spectrometer.
• Screened 99 microbial variants producing a sesquiterpenoid at 1.3 injections/min.

Main Results:

• Achieved high-throughput screening of terpene-producing microbial variants.
• Demonstrated no observable carryover between samples.
• Obtained results comparable to traditional gas chromatography-flame ionization detection (GC-FID) with a correlation coefficient of 0.96.
• Identified parameters for further throughput enhancement.

Conclusions:

• The developed FIA-APCI-Q-TOF-MS method is a powerful tool for rapid terpene screening.
• This method significantly reduces analysis time compared to GC-FID.
• It facilitates faster strain engineering for improved terpene production in industrial biotechnology.