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苯丙氨酸和谷氨酸联合处理对大豆芽菜中5-甲基四氢叶酸富集的影响 预览

Effects of combined treatment of phenylalanine and glutamate on the accumulation of 5-methyltetrahydrofolate in soybean sprouts
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摘要 [目的]本文旨在研究苯丙氨酸与谷氨酸对大豆芽菜中5-甲基四氢叶酸(5-CH3-THF)富集的影响及其机制。[方法]通过单因素及全排列试验优化出苯丙氨酸与谷氨酸的最适处理浓度,采用HPLC法测定5-CH3-THF和对氨基苯甲酸(pABA)含量,采用氨基酸自动分析仪测定氨基酸组成,并通过荧光定量PCR测定叶酸合成相关基因相对表达量。[结果]苯丙氨酸与谷氨酸处理均显著增加大豆芽菜中5-CH3-THF含量,其含量随苯丙氨酸浓度的增加呈先升后降趋势,且苯丙氨酸浓度为3mmol· L^-1时5-CH3-THF含量最高;而当谷氨酸浓度大于0.5mmol· L^-1时,谷氨酸各浓度处理之间差异不显著。二者联合处理结果表明:3mmol· L^-1苯丙氨酸和1.0mmol· L^-1谷氨酸为最佳联合施用浓度,在此条件下,大豆芽菜中5-CH3-THF含量最高,比对照增加55.50%;pABA含量比对照增加134.49%;氨基酸代谢活跃,其中甲硫氨酸、甘氨酸及组氨酸等含量均增加;叶酸合成相关基因的相对表达量提高,羟甲基二氢蝶呤焦磷酸化-二氢喋酸合成酶(HPPK-DHPS)、二氢叶酸还原酶(DHFR)、多聚谷氨酸合成酶(FPGS)等基因的相对表达量分别为对照的4.91、5.58、3.59倍。[结论]苯丙氨酸与谷氨酸联合处理大豆芽菜可促进氨基酸代谢,增大叶酸的需要量,提高叶酸合成相关基因相对表达量,促进中间底物pABA的积累,从而使5-CH3-THF得以富集。 [Objectives] The aim of this study was to investigate the effect of phenylalanine and glutamate treatment on the enrichment of 5-methyltetrahydrofolate(5-CH 3-THF)and its mechanism in soybean sprouts.[Methods] The concentration of phenylalanine and glutamate was optimized by the single-factor and permutation experiment. The content of 5-CH 3-THF and p-aminobenzoic acid( pABA )was determined by HPLC. The amino acid composition was determined by amino acid automatic analyzer and the relative expression levels of the folate synthesis-related genes were determined by real-time fluorescence quantitative PCR.[Results] Both phenylalanine and glutamate treatment significantly increased the 5-CH 3-THF content in soybean sprouts,the content of 5-CH 3-THF increased first and then decreased with the increase of phenylalanine concentration,and reached the maximum when the concentration of phenylalanine was 3 mmol · L^-1 . However,when the concentration of glutamate was greater than 0.5 mmol · L^-1 ,the difference between the different treatments was not significant. The results showed that 3 mmol · L^-1 phenylalanine combined with 1.0 mmol · L^-1 glutamate was the best application concentration. Under this condition,the content of 5-CH 3-THF in soybean sprouts was the highest,which was 55.50% higher than that of the control,and the pABA content increased by 134.49%. The amino acid metabolism was active ,and the contents of methionine,glycine and histidine increased. The relative expression level of folate synthesis-related genes increased ,including hydroxymethyldihydropterin pyrophosphokinase-dihydropteroate synthase(HPPK-DHPS),dihydrofolate reductase (DHFR),and folypolyglutamate synthase(FPGS),which were 4.91,5.58,3.59 times of control individually.[Conclusions] The combined treatment with phenylalanine and glutamate on soybean sprouts could promote amino acid metabolism and increase the requirement of folate,and thereby increased the relative expression of folate synthesis-related genes,promoted the accumulation of int
作者 任文芳 王沛 常建伟 张钰漩 杨润强 顾振新 REN Wenfang;WANG Pei;CHANG Jianwei;ZHANG Yuxuan;YANG Runqiang;GU Zhenxin(College of Food Science and Technology,Nanjing Agricultural University,Nanjing 210095,China)
出处 《南京农业大学学报》 CAS CSCD 北大核心 2019年第4期759-766,共8页 Journal of Nanjing Agricultural University
基金 江苏省农业重点研发计划项目(BE2016373).
关键词 大豆芽菜 苯丙氨酸 谷氨酸 5-甲基四氢叶酸(5-CH3-THF) 富集 soybean sprouts phenylalanine glutamate 5-methyltetrahydrofolate(5-CH3-THF) enrichment
作者简介 任文芳,硕士研究生;通信作者:顾振新,教授,博导,研究方向为生物技术与功能食品,E-mail:guzx@njau.edu.cn。
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