Mn~(2+)对植物乳杆菌影响的代谢组学分析
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摘要
利用代谢组学研究分析技术,对比研究了锰离子缺乏对植物乳杆菌发酵过程中代谢途径变化的影响。采用气-质连用技术(GC-MS)测定菌体胞内代谢物,通过主成分分析(PCA)对不同处理下的乳酸菌胞内代谢组轮廓差异进行分析,采用偏最小二乘分析(PLS)识别不同处理的差异性代谢物。结果表明,锰离子缺乏对植物乳杆菌胞内能量代谢、脂肪酸代谢、氨基酸代谢等均有显著影响。富集分析和代谢途径分析也表明锰离子具有调节胞内糖代谢和氨基酸代谢的作用。锰离子缺乏在乳酸菌发酵前期和中期会显著提高胞内氨基酸的含量,同时胞内三羧酸循环的效率因锰离子的缺乏而大幅度下降,进而影响乳酸菌的发酵活力。
The relationship between manganese and intracellular metabolites in Lactobacillus plantarum was exploited in the metabolism level by using GC-MS. The metabolic profiling of L. plantarum was compared by principal components analysis(PCA), and the characteristic metabolites were recognized by partial least squares(PLS) method. The results showed that Mn2+could impact many metabolic reactions including energy metabolism, fatty acid metabolism and amino acid metabolism. GO analysis and KEGG metabolic pathway analysis revealed that Mn2+may act as a ‘metabolic switch'for glycometabolism and amino acid metabolism. Concentrations of intracellular amino acids were improved by the lack of manganese in the early and later stage of fermentation, and the acids was dropped when the fermentation was nearly finished. The efficiency of tricarboxylic acid cycle will be reduced by the lack of Mn2 +, which may be a reason for the drop of fermentative activity in L. plantarum.
The relationship between manganese and intracellular metabolites in Lactobacillus plantarum was exploited in the metabolism level by using GC-MS. The metabolic profiling of L. plantarum was compared by principal components analysis(PCA), and the characteristic metabolites were recognized by partial least squares(PLS) method. The results showed that Mn2+could impact many metabolic reactions including energy metabolism, fatty acid metabolism and amino acid metabolism. GO analysis and KEGG metabolic pathway analysis revealed that Mn2+may act as a ‘metabolic switch'for glycometabolism and amino acid metabolism. Concentrations of intracellular amino acids were improved by the lack of manganese in the early and later stage of fermentation, and the acids was dropped when the fermentation was nearly finished. The efficiency of tricarboxylic acid cycle will be reduced by the lack of Mn2 +, which may be a reason for the drop of fermentative activity in L. plantarum.
引文
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[19]常艳芬,宫文霞,郑艳红,等.黄芩醇提物干预D-半乳糖致衰老大鼠的尿液代谢组学研究[J].药学学报, 2016, 51(1):86-92.
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[23]周宏伟,谭凤仪,钟音,等.代谢组学及其在微生物领域的研究进展[J].分析化学, 2007, 35(2):309-314.
[2] HANSEN E B. Commercial bacterial starter cultures for fermented foods of the future[J]. International Journal of Food Microbiology, 2002, 78(1/2):119-131.
[3] UDOMSIL N, SHU C, RODTONG S, et al. Quantification of viable bacterial starter cultures of Virgibacillus sp. and Tetragenococcus halophilus in fish sauce fermentation by real-time quantitative PCR[J].Food Microbiology, 2016, 57(8):54-62.
[4] BENSCH G, R譈GER M, WASSERMANN M, et al.Flow cytometric viability assessment of lactic acid bacteria starter cultures produced by fluidized bed drying[J]. Applied Microbiology&Biotechnology,2014, 98(11):4897-4909.
[5]叶雷,陈庆森,阎亚丽,等.流式细胞术快速检测直投式发酵剂菌体活力[J].食品科学, 2014, 35(10):139-144.
[6] SPINNLER H E, CORRIEU G. Automatic method to quantify starter activity based on ph measurement[J]. Journal of Dairy Research, 1989, 56(5):755-764.
[7]叶雪飞,阮辉,冯石开,等.发酵乳杆菌增殖培养基营养因子优化研究[J].食品科学, 2010, 31(5):194-196.
[8]柏建玲,莫树平,郑婉玲,等.乳酸菌增菌培养基的营养因子优化[J].食品与发酵工业, 2007, 33(2):79-81.
[9]程新,董英,苏萍,等.以菊芋为主要营养基质的高活力乳酸菌发酵剂制备[J].食品与发酵工业,2014, 40(12):66-71.
[10] CHENG X, DONG Y, SU P, et al. Improvement of the fermentative activity of lactic acid bacteria starter culture by the addition of Mn2+[J]. Applied Biochemistry&Biotechnology, 2014, 174(5):1752-1760.
[11]郭兴华.益生菌基础与应用[M].北京:科学技术出版社, 2002.
[12] FAIJES M, MARS A E, SMID E J.. Comparison of quenching and extraction methodologies for metabolome analysis of Lactobacillus plantarum[J]. Microbial Cell Factories, 2007, 6(1):27.
[13]赵延胜,肖香,周兴华,等.冻干保护剂影响植物乳杆菌代谢途径的研究[J].现代食品科技, 2016,32(8):103-108.
[14] XIA J, SINELNIKOV I V, HAN B, et al. MetaboAnalyst 3.0-making metabolomics more meaningful[J].Nucleic Acids Research, 2015, 43(W1):251-257.
[15] PENG J B, JIA H M, XU T, et al. A1H NMR based metabonomics approach to progression of coronary atherosclerosis in a rabbit model[J]. Process Biochemistry, 2011, 46(12):2240-2247.
[16]张瑞兴,刘舒,皮子凤,等.汞离子对细胞代谢通路影响的代谢组学[J].高等学校化学学报, 2014,35(06):1146-1151.
[17]柯朝甫,张涛,武晓岩,等.代谢组学数据分析的统计学方法[J].中国卫生统计, 2014, 31(2):357-359.
[18]米霞,李震宇,秦雪梅,等.基于NMR代谢组学技术的不同性状款冬花药材的化学比较[J].药学学报, 2013, 48(11):1692-1697.
[19]常艳芬,宫文霞,郑艳红,等.黄芩醇提物干预D-半乳糖致衰老大鼠的尿液代谢组学研究[J].药学学报, 2016, 51(1):86-92.
[20] VINAYAVEKHIN N, MAHIPANT G, VANGNAI A S, et al. Untargeted metabolomics analysis revealed changes in the composition of glycerolipids and phospholipids in Bacillus subtilis under 1-butanol stress[J]. Applied Microbiology&Biotechnology, 2015,99(14):1-13.
[21] XIA J, WISHART D S. Web-based inference of biological patterns, functions and pathways from metabolomic data using MetaboAnalyst[J]. Nature Protocols, 2011, 6(6):743-760.
[22]孙茂成,李艾黎,霍贵成,等.乳酸菌代谢组学研究进展[J].微生物学通报, 2012, 39(10):1499-1505.
[23]周宏伟,谭凤仪,钟音,等.代谢组学及其在微生物领域的研究进展[J].分析化学, 2007, 35(2):309-314.