研究简报 Short Communications

热带假丝酵母固体发酵代谢物提取方法的比较分析

  • 聂存喜 ,
  • 张文举 ,
  • 闫理东 ,
  • 蒋粒薪 ,
  • 马贵军
展开
  • 1. 石河子大学动物科技学院,石河子 832003;
    2. 新疆天康畜牧生物技术股份有限公司 生物添加剂分公司,乌鲁木齐 830011

收稿日期: 2012-02-01

  网络出版日期: 2012-08-03

基金资助

国家自然科学基金项目(30960258);石河子大学科技成果转化引导资金专项(kjkfyzh2009-cgzh01)

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

摘要

本试验比较了几种提取微生物固体发酵代谢物的方法,旨在筛选出一种尽可能多保留了代谢物的方法。试验采用热带假丝酵母固体发酵棉粕源底物,以液相色谱串联质谱(LC-MS)为分析平台,分别采用50%冷甲醇溶液(甲醇法)、酸性甲醇-乙腈溶液(酸性甲醇-乙腈法)和50%乙腈溶液(乙腈法)提取发酵代谢物。结果表明:采用酸性甲醇-乙腈法、乙腈法和甲醇法分别提取出1 831、1 522和1 262种代谢物,提取效率表现为酸性甲醇-乙腈法>甲醇法>乙腈法。本试验中3种方法相比,酸性甲醇-乙腈法提取的代谢物种类最多,提取效率最佳,是微生物固体发酵代谢物提取的有效方法。

本文引用格式

聂存喜 , 张文举 , 闫理东 , 蒋粒薪 , 马贵军 . 热带假丝酵母固体发酵代谢物提取方法的比较分析[J]. 动物营养学报, 2012 , 24(8) : 1610 -1614 . DOI: 10.3969/j.issn.1006-267x.2012.08.028

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.

参考文献

[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.

文章导航

/