硫酸铵提高裂殖壶菌不饱和脂肪酸积累的代谢组学研究

引用本文: 陈胜兰, 赵晓艳, 任路静, 纪晓俊, 黄和. 硫酸铵提高裂殖壶菌不饱和脂肪酸积累的代谢组学研究[J]. 分析化学, 2016, 44(5): 685-692. doi: 10.11895/j.issn.0253-3820.150948 shu

Citation: CHEN Sheng-Lan, ZHAO Xiao-Yan, REN Lu-Jing, JI Xiao-Jun, HUANG He. Metabonomics Study on Enhancement of Unsaturated Fatty Acids Accumulation by Ammonium Sulfate in Schizochytrium sp.[J]. Chinese Journal of Analytical Chemistry, 2016, 44(5): 685-692. doi: 10.11895/j.issn.0253-3820.150948 shu

硫酸铵提高裂殖壶菌不饱和脂肪酸积累的代谢组学研究

南京工业大学, 生物与制药工程学院, 南京 211816
基金项目:
本研究系国家863计划(No.2014AA021701),国家自然科学基金(No.21306085),高等学校博士学科点专项科研基金(No.20133221120008)资助

摘要: 采用基于气相色谱-质谱(GC-MS)联用技术的代谢组学方法,通过分析裂殖壶菌胞内代谢物的变化,研究(NH₄)₂SO₄提高裂殖壶菌不饱和脂肪酸积累的作用机制。利用GC-MS技术对添加0,1.5,2.5和3.5 g/L (NH₄)₂SO₄的发酵样品进行分析,结合K-Means算法对数据进行聚类分析,得到胞内代谢物的热图。主成分分析(PCA)和偏最小二乘判别分析(OPLS-DA)得分图可将不同添加条件下裂殖壶菌胞内物质明显区分。运用PLS-DA载荷图及模型的变量重要性因子(VIP)值,发现了7种代谢物可作为区别4种不同条件下的生物标志物。经NIST和Wiley谱库检索,它们分别为葡萄糖、肌醇、甘露醇、果糖、磷酸、乙醇胺、乙胺。代谢途径分析结果表明,(NH₄)₂SO₄是通过弱化糖酵解途径和乙酸合成支路,提高苹果酸酶的活力,强化氨基酸代谢和NADPH的积累,从而提高不饱和脂肪酸积累。

关键词:

English

Metabonomics Study on Enhancement of Unsaturated Fatty Acids Accumulation by Ammonium Sulfate in Schizochytrium sp.

College of Biotechnology and Pharmaceutical Engineering, Nanjing Tech University, Nanjing 211816, China
Received Date: 25 August 2015
Revised Date: 18 January 2016
Fund Project:
This work was supported by the National Natural Science Foundation of China (No. 21306085)

Abstract: A metabonomics method based on gas chromatography-mass spectrometry (GC-MS) was established to analyze the changes of intracellular metabolites and study the mechanism of ammonium sulfate enhancing unsaturated fatty acids accumulation in Schizochytrium sp. Metabolites of cells cultivated at 0, 1.5, 2.5 and 3.5 g/L (NH₄)₂SO₄ were analyzed at different times. Heat map was obtained with K-Means algorithm for data clustering analysis. Principal component analysis (PCA) and partial least-squares discrimination analysis (OPLS-DA) were employed for pattern recognition, and an obvious distinction among different conditions was achieved. According to the LS-DA load diagram and variable important factor (VIP) value, 7 biomarkers were determined with significant differences between four different conditions, which were identified as glucose, inositol, mannitol, fructose, phosphoric acid, ethylamine and ethanolamine by searching in NIST and Wiley database. Metabolic pathway analysis indicated that the metabolic flux toward the Embden-Meyerh of-Parnas (EMP) pathway and acetic acid synthesis branch were weakened by ammonium sulfate. Meanwhile, malic enzyme activity increased and the accumulation of NADPH was also enhanced. These might be the reason for the improvement of polyunsaturated fatty acids.

Key words:

1. [1]
  Koek M M, Jellema R H, Van der Greef J, Tas A C, Hankemeier T. Metabolomics, 2011,7(3):307-328
2. [2]
  XIE Yue-Sheng, PAN Gui-Xiang, GAO Xiu-Mei, LIU Chang-Xiao. Chinese J. Anal. Chem., 2006,34(11):1644-1648 谢跃生, 潘桂湘, 高秀梅, 刘昌孝.分析化学,2006,34(11):1644-1648
3. [3]
  SHI Dong-Dong, WANG Gui-Ming, KUANG Yuan-Yuan, PENG Zhang-Xiao, WANG Yan, GU Xue, YAN Chao. Chinese J. Anal.Chem., 2014,42(8):1088-1093 史栋栋, 王桂明, 况媛媛, 彭章晓, 王彦, 谷雪, 阎超.分析化学,2014,42(8):1088-1093
4. [4]
  Timmins A, Macpherson E J, Ackman R G. Food Chem., 2000,70(3):425-426
5. [5]
  Horrocks L A, Yeo Y K. Pharmacol. Res., 1999,40(3):211-225
6. [6]
  Giusto N, Pasquare S, Salvador G, Castagnet P, Roque M, Ilincheta de Boschero M. Prog. Lipid Res., 2000,39(4):315-391
7. [7]
  Ren L J, Sun L N, Zhuang X Y, Qu L, Ji X J, Huang H. Bioprocess. Biosyst. Eng., 2014,37(5):865-872
8. [8]
  Sun L, Ren L, Zhuang X, Ji X, Yan J, Huang H. Bioresour. Technol., 2014,159:199-206
9. [9]
  Ling X, Guo J, Liu X, Zhang X, Wang N, Lu Y, Ng I S. Bioresour. Technol., 2015,184:139-147
10. [10]
  Ren L J, Sun G N, Ji X J, Hu X C, Huang H. Bioresour. Technol., 2014,157:107-113
11. [11]
  Ren L J, Ji X J, Huang H, Qu L, Feng Y, Tong Q Q, Ouyang P K. Appl. Microbiol. Biotechnol., 2010, 87(5):1649-1656
12. [12]
  Hsu R Y, Lardy H A. Methods Enzymol., 1969,13:230-235
13. [13]
  Fritz C, Mueller C, Matt P, Feil R, Stitt M. Plant. Cell. Environ., 2006,29(11):2055-2076
14. [14]
  Li J, Ren L J, Sun G N, Qu L, Huang H. OMICS, 2013,17(5):269-281
15. [15]
  Ratledge C. Biochimie, 2004,86(11):807-815
16. [16]
  Zhang Y, Adams I P, Ratledge C. Microbiology, 2007,153(7):2013-2025
17. [17]
  Pearson C J, Jacobs B C. Aust. J. Agr. Res., 1987,38(6):1001-1009
18. [18]
  Jackowski S, Rock C O. Biochim. Biophys. Acta, Mol. Cell Biol. Lipids, 2013,1831(3):469-470
19. [19]
  Turk M, Mejanelle L, Sentjurc M, Grimalt J O, Gunde-Cimerman N, Plemenitas A. Extremophiles, 2004,8(1):53-61
20. [20]
  Siniossoglou S. Biochim. Biophys. Acta, Mol. Cell Biol. Lipids, 2013,1831(3):575-581
21. [21]
  Jennings D H. Adv. Appl. Microbiol., 1985,25:149-193
22. [22]
  Nikawa J, Yamashita S. Biochim. Biophys. Acta, Mol. Cell Biol. Lipids, 1997,1348(1-2):173-178
23. [23]
  Ding M Z, Cheng J S, Xiao W H, Qiao B, Yuan Y J. Metabolomics, 2009,5(2):229-238

扫一扫看文章

计量

PDF下载量: 0
文章访问数: 1098
HTML全文浏览量: 109

通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142

硫酸铵提高裂殖壶菌不饱和脂肪酸积累的代谢组学研究

English

Metabonomics Study on Enhancement of Unsaturated Fatty Acids Accumulation by Ammonium Sulfate in Schizochytrium sp.

CCS Chemistry：嵌段共聚物自组装成 “三明治”二维有机材料，实现纳米级压力传感功能

CCS Chemistry：颜徐州、鲍哲南：你的皮肤可能是一片SPMs

CCS Chemistry：工作高效不疲劳，只须让催化剂动起来

CCS Chemistry：静电组装导向聚合- 聚电解质纳米凝胶量化制备新策略

CCS Chemistry：金黄色葡萄球菌醛基脱氢酶是如何识别特异性底物的？

计量

通讯作者: 陈斌, bchen63@163.com

中国化学会秘书处

硫酸铵提高裂殖壶菌不饱和脂肪酸积累的代谢组学研究

English

Metabonomics Study on Enhancement of Unsaturated Fatty Acids Accumulation by Ammonium Sulfate in Schizochytrium sp.

CCS Chemistry：嵌段共聚物自组装成 “三明治”二维有机材料，实现纳米级压力传感功能

CCS Chemistry：颜徐州、鲍哲南： 你的皮肤可能是一片SPMs

CCS Chemistry：工作高效不疲劳，只须让催化剂动起来

CCS Chemistry：静电组装导向聚合- 聚电解质纳米凝胶量化制备新策略

CCS Chemistry：金黄色葡萄球菌醛基脱氢酶是如何识别特异性底物的？

计量

文章相关

通讯作者: 陈斌, bchen63@163.com

中国化学会秘书处

CCS Chemistry：颜徐州、鲍哲南：你的皮肤可能是一片SPMs