TY - JOUR
T1 - Fast filtration with a vacuum manifold system as a rapid and robust metabolome sampling method for Saccharomyces cerevisiae
AU - Yun, Eun Ju
AU - Seo, Seung Oh
AU - Kwak, Suryang
AU - Oh, Eun Joong
AU - Lee, Sun Hee
AU - Jin, Yong Su
AU - Kim, Kyoung Heon
N1 - Publisher Copyright:
© 2021 Elsevier Ltd
PY - 2021/11
Y1 - 2021/11
N2 - For metabolome sample preparation, rapid quenching of cellular metabolism without severe loss of intracellular metabolites is important. Here, we comparatively evaluated the two sampling methods, cold methanol quenching and fast filtration, for one of the most important industrial microorganisms, yeast Saccharomyces cerevisiae. The latter, using a vacuum manifold system capable of processing up to twenty samples simultaneously, was developed here. The results showed that the cold methanol quenching method resulted in severe losses of intracellular metabolites but that sugars and sugar phosphates were highly preserved. In comparison, the fast filtration method better preserved most metabolites with less extracellular contamination. Our results suggested that the fast filtration method with a vacuum manifold system can be widely applied to yeast metabolome analysis due to its simplicity, robustness, and lack of significant intracellular metabolite loss.
AB - For metabolome sample preparation, rapid quenching of cellular metabolism without severe loss of intracellular metabolites is important. Here, we comparatively evaluated the two sampling methods, cold methanol quenching and fast filtration, for one of the most important industrial microorganisms, yeast Saccharomyces cerevisiae. The latter, using a vacuum manifold system capable of processing up to twenty samples simultaneously, was developed here. The results showed that the cold methanol quenching method resulted in severe losses of intracellular metabolites but that sugars and sugar phosphates were highly preserved. In comparison, the fast filtration method better preserved most metabolites with less extracellular contamination. Our results suggested that the fast filtration method with a vacuum manifold system can be widely applied to yeast metabolome analysis due to its simplicity, robustness, and lack of significant intracellular metabolite loss.
KW - Cold methanol quenching
KW - Fast filtration
KW - Gas chromatography–mass spectrometry
KW - Metabolome sample preparation
KW - Metabolomics
KW - Saccharomyces cerevisiae
KW - Sampling method
UR - http://www.scopus.com/inward/record.url?scp=85113187496&partnerID=8YFLogxK
U2 - 10.1016/j.procbio.2021.08.012
DO - 10.1016/j.procbio.2021.08.012
M3 - Article
AN - SCOPUS:85113187496
SN - 1359-5113
VL - 110
SP - 195
EP - 200
JO - Process Biochemistry
JF - Process Biochemistry
ER -