Expanded metabolite coverage of Saccharomyces cerevisiae extract through improved chloroform/methanol extraction and tert-butyldimethylsilyl derivatization
Journal article, 2015

We present an improved extraction and derivatization protocol for GC-MS analysis of amino/non-amino acids in Saccharomyces cerevisiae. Yeast cells were extracted with chloroform: aqueous-methanol (1:1, v/v) and the resulting non-polar and polar extracts combined and dried for derivatization. Polar and non-polar metabolites were derivatized using tert-butyldimethylsilyl (t-BDMS) dissolved in acetonitrile. Using microwave treatment of the samples, the derivatization process could be completed within 2 h (from >20 h of the conventional method), providing fully derivatized metabolites that contain multiple derivatizable organic functional groups. This results in a single derivative from one metabolite, leading to increased accuracy and precision for identification and quantification of the method. Analysis of combined fractions allowed the method to expand the coverage of detected metabolites from polar metabolites i.e. amino acids, organic acids and non-polar metabolites i.e. fatty alcohols and long-chain fatty acids which are normally non detectable. The recoveries of the extraction method was found at 88 ± 4%, RSD, N = 3 using anthranilic acid as an internal standard. The method promises to be a very useful tool in various aspects of biotechnological applications i.e. development of cell factories, metabolomics profiling, metabolite identification, 13C-labeled flux analysis or semi-quantitative analysis of metabolites in yeast samples.

Metabolomics

Extraction

Derivatization

Saccharomyces cerevisiae

Author

Sakda Khoomrung

Chalmers, Biology and Biological Engineering, Systems and Synthetic Biology

Jose Luis Martinez Ruiz

Chalmers, Biology and Biological Engineering, Systems and Synthetic Biology

Stefan Tippmann

Chalmers, Biology and Biological Engineering, Systems and Synthetic Biology

Suwanee Jansa-Ard

Chalmers, Biology and Biological Engineering, Systems and Synthetic Biology

Marieke Buffing

Chalmers, Biology and Biological Engineering, Systems and Synthetic Biology

Raffaele Nicastro

Chalmers, Biology and Biological Engineering, Systems and Synthetic Biology

Jens B Nielsen

Chalmers, Biology and Biological Engineering, Systems and Synthetic Biology

Analytical Chemistry Research

2214-1812 (eISSN)

Vol. 6 9-16

Areas of Advance

Life Science Engineering (2010-2018)

Subject Categories

Bioinformatics and Systems Biology

DOI

10.1016/j.ancr.2015.10.001

More information

Created

10/7/2017