Metabolic profiling of Klebsiella oxytoca: evaluation of methods for extraction of intracellular metabolites using UPLC/Q-TOF-MS

Applied Biochemistry and Biotechnology
Changhun ParkJinwon Lee

Abstract

The global pool of intracellular metabolites is a reflection of all the metabolic functions of an organism. In the absence of in situ methods capable of directly measuring metabolite pools, intracellular metabolite measurements need to be performed after an extraction procedure. In this study, we evaluated the optimization of technologies for generation of a global metabolomics profile for intracellular metabolites in Klebsiella oxytoca. Intracellular metabolites of K. oxytoca were extracted at the early stationary phase using six different common extraction procedures, including cold methanol, boiling ethanol, methanol/chloroform combinations, hot water, potassium hydroxide, and perchloric acid. The metabolites were subsequently collected for further analysis, and intracellular metabolite concentration profiles were generated using ultra-performance liquid chromatography/quadrupole time-of-flight mass spectrometry. During analysis, the stability of metabolites extracted using cold methanol was clearly higher than that obtained by other extraction methods. For the majority of metabolites, extracts generated in this manner exhibited the greatest recovery, with high reproducibility. Therefore, the use of cold ethanol was the best...Continue Reading

References

Apr 1, 1975·Journal of Bacteriology·F M Harold, E Spitz
Nov 15, 1986·Analytical Biochemistry·J C ShryockR M Berne
Dec 1, 1972·Biochimica Et Biophysica Acta·A M Cook, C A Fewson
Jan 1, 1995·The Biochemical Journal·N MoreH H Petach
Jul 19, 2000·Biochimica Et Biophysica Acta·J M BraggerR M Daniel
Oct 17, 2001·Applied Microbiology and Biotechnology·M A HansC Wittmann
Oct 23, 2001·Biotechnology and Bioengineering·H C LangeJ J Heijnen
Feb 12, 2002·Journal of the American Society for Mass Spectrometry·Seetharaman VaidyanathanRoyston Goodacre
Jul 10, 2002·Biomolecular Engineering·Arne BuchholzRalf Takors
Feb 11, 2003·Analytical Biochemistry·Ram Prasad Maharjan, Thomas Ferenci
Oct 22, 2005·Yeast·Silas G Villas-BôasJens Nielsen
Jun 6, 2006·Mass Spectrometry Reviews·Sílvia Lacorte, Amadeo R Fernandez-Alba
Aug 22, 2006·Mass Spectrometry Reviews·Katja DettmerBruce D Hammock
Feb 14, 2007·Journal of Mass Spectrometry : JMS·Guodong ChenUrooj A Mirza
May 5, 2007·Biotechnology Letters·Julia HillerDirk Weuster-Botz
Jan 24, 2008·Molecular BioSystems·Yoshiaki OhashiTomoyoshi Soga
Mar 14, 2008·Journal of Separation Science·Guoxiang XieWei Jia
Aug 21, 2008·Nature Protocols·Bryson D BennettJoshua D Rabinowitz
Aug 6, 2009·Analytical Chemistry·André B CanelasJoseph J Heijnen
Jul 24, 2010·Bulletin of Environmental Contamination and Toxicology·H B YaoC H Wang
Feb 22, 2012·Trends in Analytical Chemistry : TRAC·Jun Feng XiaoHabtom W Ressom

❮ Previous
Next ❯

Citations

Sep 14, 2012·Applied Biochemistry and Biotechnology·Aihua ZhangXijun Wang
Sep 2, 2014·Metabolomics : Official Journal of the Metabolomic Society·Robert M HowlettDavid J Kelly
Oct 5, 2018·PloS One·Charulata B PrasannanPramod P Wangikar
Nov 16, 2018·PloS One·Claudio FoschiMaria Carla Re
Apr 7, 2015·Prikladnaia biokhimiia i mikrobiologiia·R K PuzanskiĭM F Shishova
May 20, 2014·Applied Biochemistry and Biotechnology·Aihua ZhangXijun Wang
Mar 5, 2019·Metabolomics : Official Journal of the Metabolomic Society·David J BealeDaniel A Dias
Dec 13, 2016·Medical Oncology·Xiaoqian WangDavid T W Wong

❮ Previous
Next ❯

Related Concepts

Related Feeds

Acute viral rhinopharyngitis

Acute viral rhinopharyngitis, also known as "common cold", is an acute, self-limiting viral infection of the upper respiratory tract involving the nose, sinuses, pharynx and larynx. Discover the latest research on acute viral rhinopharyngitis here.