High-Throughput Metabolite Analysis of Unicellular Microalgae by Orthogonal Hybrid Ionization Label-Free Mass...

Area of Science:

• Biotechnology
• Analytical Chemistry
• Microalgae Research

Background:

• Metabolite analysis in microalgae is crucial for metabolic engineering.
• Current mass spectrometry (MS) methods for single-cell microalgae analysis have limitations in throughput and detectable substance polarity.
• High-throughput screening of high-performance microalgae is challenging.

Purpose of the Study:

• To develop a novel method for rapid, high-throughput metabolite detection in single microalgae.
• To enable detailed analysis of microalgae metabolism under various conditions.
• To support the rational design of metabolic engineering strategies for high-value product biosynthesis.

Main Methods:

• Development and application of plasma-assisted label-free mass cytometry (PACyESI-MS).
• Utilizing orthogonal hybrid ionization and high-throughput MS analysis for metabolite detection.
• Analysis of metabolite changes in *Chlamydomonas reinhardtii* and *Haematococcus pluvialis* under nitrogen deficiency.

Main Results:

• PACyESI-MS achieved a cell detection throughput of up to 52 cells/min.
• Simultaneous detection of dozens of critical primary and secondary metabolites (pigments, lipids, energy metabolites) in single microalgae.
• Discrimination of *Chlamydomonas* under different nutrient conditions based on single-cell metabolite profiles.
• Investigation of the relationship between astaxanthin accumulation and primary metabolites in *Haematococcus*.

Conclusions:

• PACyESI-MS is a powerful tool for rapid, label-free, single-cell metabolite analysis in microalgae.
• The method effectively detects flexible metabolite changes under varying nutritional conditions and during high-value product synthesis.
• PACyESI-MS is expected to significantly advance the design of metabolic engineering strategies for high-performance microalgae factories.