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Rapid and accurate identification of yeast subspecies by MALDI-MS combined with a cell membrane disruption reagent

Nan Zhao1, Wei Guo2, Jiarui Li1

  • 1State Key Laboratory of Supramolecular Structure and Materials, College of Chemistry, Jilin University, Changchun 130012, China.

Food Chemistry
|June 21, 2024
PubMed

Insights

A new reagent, C18-G1, effectively disrupts fungal cell walls for improved matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) analysis. This method enhances fungal identification and discrimination, overcoming previous limitations.

Area of Science:

  • Microbiology
  • Analytical Chemistry
  • Biochemistry

Background:

  • Matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) is a key technique for microbial identification.
  • The robust cell walls of fungi present a significant challenge for direct MALDI-MS analysis.
  • Existing fungal pretreatment methods for MALDI-MS are often suboptimal for comprehensive analysis.

Purpose of the Study:

  • To develop and evaluate a novel fungal cell membrane disruption reagent for enhanced MALDI-MS.
  • To improve the sensitivity and range of detectable biomarkers from fungal samples.
  • To enable more accurate fungal identification and subspecies discrimination using MALDI-MS.

Main Methods:

  • Utilized Saccharomyces cerevisiae as a model organism for fungal analysis.
  • Applied a new fungal cell membrane disruption reagent, C18-G1, prior to MALDI-MS.
  • Employed MS/MS fragmentation and database searching to identify specific proteins.

Main Results:

  • The C18-G1 reagent significantly increased the intensity and m/z range of detectable peaks from S. cerevisiae compared to traditional methods.
  • A differential peak at m/z 4993 was identified, corresponding to a protein unique to a specific S. cerevisiae subspecies.
  • The method successfully discriminated between four commercial yeast species.

Conclusions:

  • The C18-G1 reagent offers a superior approach for fungal cell disruption in MALDI-MS.
  • This advancement facilitates more precise fungal identification, subspecies differentiation, and biomarker discovery.
  • The developed method holds potential for broader applications in yeast and fungal analysis.
Keywords:
MALDI-MSS. cerevisiaeYeast disruption reagentYeast identification

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