Molecular Nutrition

A Comparison of Extraction Methods for Metabolites in Solid Fermentation with Candida tropicalis

  • NIE Cunxi ,
  • ZHANG Wenju ,
  • YAN Lidong ,
  • JIANG Lixin ,
  • MA Guijun
Expand
  • 1. College of Animal Science and Technology, Shihezi University, Shihezi 832003, China;
    2. Brand of Bio-Additive, Xinjiang Tecon Animal Science Bio-Technology Co. , Ltd. , Urumqi 830011, China

Received date: 2012-02-01

  Online published: 2012-08-03

Abstract

In order to find a good extraction method that can keep metabolites as many as possible, several extraction methods for solid fermentation metabolites were compared in this study. Substrates with cottonseed meal were fermented by Candida tropicalis, and the analysis of metabolites was carried out using the liquid chromatographic-mass spectrometry (LC-MS) platform. Three different extraction solutions, including 50% cold methanol solution (MeOH method), acidic methanol-acetonitrile solution (AMF method) and 50% acetonitrile solution (MeCN method), were used. The results showed that the number kinds of metabolites extracted by methods of AMF, MeCN and MeOH were 1 831, 1 522 and 1 262, respectively; extraction efficiency was shown as AMF method>MeOH method>MeCN method. Among the three methods in this experiment, AMF method is an effective method for metabolites extraction in solid fermentation, and acts better on both kinds of metabolites and extraction efficiency.

Cite this article

NIE Cunxi , ZHANG Wenju , YAN Lidong , JIANG Lixin , MA Guijun . A Comparison of Extraction Methods for Metabolites in Solid Fermentation with Candida tropicalis[J]. Chinese Journal of Animal Nutrition, 2012 , 24(8) : 1610 -1614 . DOI: 10.3969/j.issn.1006-267x.2012.08.028

References

[1] OLDIGES M,LVTZ S,PFLUG S,et al.Metabolomics:current state and evolving methodologies and tools[J].Applied Microbiology and Biotechnology,2007,76(3):495-511.  

[2] MAHARJAN R P,FERENCI T.Global metabolite analysis:the influence of extraction methodology on metabolome profiles of Escherichia coli[J].Analytical Biochemistry,2003,313(1):145-154.  

[3] 梅辉,戴军,刘文卫,等.高效液相色谱-串联质谱法分析大肠杆菌代谢组中样品提取方法的比较[J].分析化学,2011,39(4):534-539.

[4] RABINOWITZ J D,KIMBALL E.Acidic acetonitrile for cellular metabolome extraction from Escherichia coli[J].Analytical Chemistry,2007,79(16):6167-6173.  

[5] WINDER C L,DUNN W B,SCHULER S.Global metabolic profiling of Escherichia coli cultures:an evaluation of methods for quenching and extraction of intracellular metabolites[J].Analytical Chemistry,2008,80(8):2939-2948.  

[6] TOKUOKA M,SAWAMURA N,KOBAYASHI K,et al.Simple metabolite extraction method for metabolic profiling of the solid-state fermentation of Aspergillus oryzae[J].Journal of Bioscience Bioengineering,2010,110(6):665-669.  

[7] HØJER-PEDERSEN J,SMEDSGAARD J,NIELSEN J.The yeast metabolome addressed by electrospray ionization mass spectrometry:initiation of a mass spectral library and its applications for metabolic footprinting by direct infusion mass spectrometry[J].Metabolomics,2008,4(4):393-405.  

[8] BIJLSMA S,BOBELDIJK I,VERHEIJ E R,et al.Large-scale human metabolomics studies:a strategy for date (pre-) processing and validation[J].Analytical Chemistry,2006,78(2):567-574.  

[9] CANELAS A B,RAS C,PIERICK A T,et al.Leakage-free rapid quenching technique for yeast metabolomics[J].Metabolomics,2008,4(3):226-239.  

[10] HILLER J,FRANCO-LARA E,PAPAIOANNOU V,et al.Fast sampling and quenching procedures for microbial metabolic profiling[J].Biotechnology Letters,2007,29(8):1161-1167.  

[11] SPURA J,REIMER L C,WIELOCH P,et al.A method for enzyme quenching in microbial metabolome analysis successfully applied to gram-positive and gram-negative bacteria and yeast[J].Analytical biochemistry,2009,394(2):192-201.  

[12] VILLAS-BOAS S G,HOJER-PEDERSEN J,AKESSON M,et al.Global metabolite analysis of yeast:evaluation of sample preparation methods[J].Yeast,2005,22:1155-1169.

[13] 余冰,傅娅梅,叶楠等.固态发酵对复合蛋白质饲料营养价值改善效果的研究[J].动物营养学报,2009,21(4):546-553.

Outlines

/