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低丰度^(13)C标记的酿酒酵母代谢通量分析探讨研究 被引量:2

Research on ^(13)CMFA of S.cerevisiae at Low Labeled Tracer Substrate
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摘要 通过对酿酒酵母发酵液进行低浓度的[1-13C]葡萄糖标记,并采用GC-C-IRMS测定发酵菌体中蛋白氨基酸的δ13C值,最终显示在0.5%~2%的标记浓度范围内,15种菌体蛋白氨基酸δ13C值与标记浓度呈现出很好的线性关系。证明了0.5%或者更低的浓度标记都可以用于13CMFA中,大大降低了碳同位素标记代谢流成本,为13CMFA应用于传统白酒和黄酒大生产发酵过程中的复杂代谢机理研究提供一种手段。 Stable carbon isotope compositions of proteinogenic amino acids in S.cerevisiae fermenting liquid at low labeled tracer substrate were determined by gas chromatography-combustion-isotope ratio mass spectrometry (GC-C-IRMS) in the experiments. The results suggested that, within 0.5 %-2 % labeled range, ^13C values of 15 kinds of amino acids in protein had good linear relationships with the labeled concentration. The experiments demonstrated that 0.5 % or even lower labeled concentration was feasible in ^13CMFA, which could reduce greatly the metabolic flux cost and provide a new approach to ^13CMFA application in the research on the complex metabolism of the fermentation of traditional liquor and yellow rice wine.
出处 《酿酒科技》 北大核心 2014年第3期13-15,20,共4页 Liquor-Making Science & Technology
基金 国家自然基金(31101333) 国际合作项目(2011DFA33270) 国家科技支撑计划(编号:2012BAK17B11) 质检公益项目(编号:201210104)
关键词 白酒 ^13C标记代谢通量分析 [1一^13C]葡萄糖 蛋白氨基酸 气相色谱-燃烧-稳定同位素比值质谱 Baijiu(liquor) ^13C metabolic flux analysis [ 1-^13C] glucose proteinogenic amino acids GC-C-IRMS
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参考文献9

  • 1Kelleher J K. Flux estimation using isotopic tracers: common ground for metabolic physiology and metabolic engineering[J]. Metabolic Engineering, 2001, 3(2): 100-110.
  • 2Marx A, de GraafA A, Wiechert W, et al. Determination of the fluxes in the central metabolism ofCorynebacterium glutam- icum by nuclear magnetic resonance spectroscopy combined with metabolite balancing[J].Biotechnology and Bioenginee- ring, 1996,49(2): 111-129.
  • 3Zamboni N, Fendt S, Rtihl M, et al. 13C-based metabolic flux analysis[J]. Nature Protocols, 2009,4(6): 878-892.
  • 4Wittmann C, Kim H M, Heinzle E. Metabolic network analysisoflysine producing Corynebacterium glutamicum at a miniatur- ized scale[J].Biotechnology and Bioengineering,2004,87(1): 1-6.
  • 5Saner U, Hatzimanikatis V, Bailey J E, et al. Metabolic fluxes in riboflavin-producing Bacillus sub tilis [J].Nature Biotechnology, 1997,15(5): 448-452.
  • 6Dauner M, Saner U. GC-MS analysis of amino acids rapidly provides rich information for isotopomer balancing[J]. Biotechnology Progress, 2000,16(4) : 642-649.
  • 7Wittmann C, Heinzle E. Application of MALDI-TOF MS to lysine - producing Corynebacterium glutamicum[J]. European Journal of Biochemistry, 2001,268(8): 2441-2455.
  • 8Velagapudi V R, Wittmann C, Schneider K, et al. Metabolic flux screening ofSaccharomyces cerevisiae single knockout strains on glucose and galactose supports elucidation of gene function[J]. Journal of Biotechnology, 2007,132(4):395-404.
  • 9钟其顶,李国辉,王道兵,熊正河,王敏.气相色谱-燃烧-同位素质谱仪(GC-C-IRMS)测定游离氨基酸的δ^(13)C值[J].酿酒科技,2013(9):7-10. 被引量:7

二级参考文献9

  • 1Stephanopoulos G. Metabolic fluxes and metabolic engineering [J].Metaboli Engineering, 1999,1(1): 1-11.
  • 2Varma A, Palsson B O. Metabolic flux balancing: basic concepts, scientific and practical use[J]. Nature Biotechnology, 1994,12(10) :994-998.
  • 3Sauer U, Lasko D R, Fiaux J, et al. Metabolic flux ratio analysis of genetic and environmental modulations ofEscherichia coli central carbon metabolism[J].Journal of Bacteriology, 1999,181 (21) :6679-6688.
  • 4Wiechert W. 13C metabolic flux analysis.[J]. Metabolic Enginee- ring, 2001,3(3): 195.
  • 5Szyperski T. 13C-N]VIR, MS and metabolic flux balancing inbiotechnology research[J]. Quarterly Reviews of Biophysics, 1998,31(01):41-106.
  • 6Christensen B, Karoly Gombert A, Nielsen J. Analysis of flux estimates based on 13C - labelling experiments[J]. European Journal of Biochemistry, 2002,269(11): 2795-2800.
  • 7Heinzle E, Yuan Y, Kumar S, et al. Analysis of t3C labeling enrichment in microbial culture applying metabolic tracer exper- iments using gas chromatography-combustion-isotope ratio mass spectrometry[J].Analytical Biochemistry, 2008,380(2): 202-210.
  • 8Yuan Y, Hoon Yang T, Heinzle E. 13C metabolic flux analysis for larger scale cultivation using gas chromatography-combustion- isotope ratio mass spectrometry[J].Metabolic Engineering, 2010,12(4): 392-400.
  • 9Tang Y J, Martin H G, Myers S, et al. Advances in analysis of microbial metabolic fluxes via 13C isotopic labeling[J]. Mass Spectrometry Reviews, 2009,28(2): 362-375.

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