普通橄榄和清橄榄果实游离氨基酸差异成分与谷氨酰胺代谢
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摘要
基于高效液相色谱-串联质谱法结合多元统计分析技术测定普通橄榄‘长营’和清橄榄‘清榄1号’果实游离氨基酸的差异成分并对其代谢进行分析,为研究氨基酸在普通橄榄和清橄榄风味贡献上的差异提供依据。通过主成分分析、偏最小二乘投影判别分析和显著性分析确定差异成分,并进行对象之间的相关性分析。结果表明:普通橄榄‘长营’和清橄榄‘清榄1号’果实中游离氨基酸成分存在差异,差异氨基酸为呈甜味的丝氨酸(Ser)、谷氨酰胺(Gln)和呈鲜味的谷氨酸(Glu),且三者均以‘清榄1号’含量居高,使‘清榄1号’Ser、Gln和Glu对风味的贡献大于‘长营’,与其鲜食品质一致,其中2 个品种平均含量排名为Glu>Gln>Ser,差异倍数排名为Ser>Gln>Glu,综合2个指标以Gln为重要氨基酸;‘长营’Gln含量与天冬酰胺合成酶(asparaginesynthetase,AS)呈显著(P<0.05)负相关(r=-0.792),与可溶性糖呈显著(P<0.05)负相关(r=-0.711),与蔗糖和还原糖相关性不显著(P>0.05),‘清榄1号’Gln含量与AS活性相关性不显著(P>0.05),与蔗糖呈极显著(P<0.01)正相关(r=0.953),与可溶性糖呈显著(P<0.05)正相关(r=0.791),与还原糖呈显著(P<0.05)负相关(r=-0.780),2 种橄榄Gln含量与其代谢酶和糖类的相关性存在差异;综上揭示了游离氨基酸对普通橄榄和清橄榄风味贡献存在差异,特别是Gln对2类橄榄的影响且其可能通过调控AS活性和糖代谢来实现对其含量的调控进而影响果实风味。
Using high performance liquid chromatography-tandem mass spectrometry(HPLC-MS/MS) combined with multivariate statistical analysis, fruits of the common olive ‘Changying' and the sweet olive ‘Qinglan 1 hao' were compared for differences in their free amino acid composition. Meanwhile, glutamine metabolism characteristics of the two olive varieties were analyzed. The aim was to provide the basis for understanding the contribution of amino acids to the differential flavor between common olive and sweet olive. By principal component analysis(PCA), partial least squares projection to latent structures discriminant analysis(PLS-DA) and significance analysis, the differential free amino acids between the two varieties were determined and correlation analysis was also conducted. The results showed that there was a difference in free amino acid composition between common olive and sweet olive with respect to the sweet amino acids serine(Ser)and glutamine(Gln) as well as umami glutamate(Glu). The three amino acids were all more abundant in ‘Qinglan 1 hao'and made greater contribution to the flavor of this variety. Their average contents were in the descending order: Glu > Gln >Ser for both varieties, and their differences between the two varieties were in descending order: Ser > Gln > Glu, overall suggesting that Gln was a more important amino acid. The Gln content of ‘Changying' was significantly negatively correlated with asparagine synthetase(AS) activity(P < 0.05) and soluble sugar with correlation coefficient(r) of-0.792 and-0.711,respectively, whereas no significant correlation with sucrose or reducing sugar was found(P > 0.05). The Gln content of‘Qinglan 1 hao' was not significantly correlated with AS activity, while it was highly significantly correlated with sucrose(P < 0.05, r = 0.953) and soluble sugar(P < 0.05, r = 0.791) and negatively correlated with reducing sugar(P < 0.05,r =-0.780). Hence, the correlations of Gln content with the metabolic enzyme activities and sugars differed between both varieties. In conclusion, free amino acids made different contributions to the flavor of common olive and sweet olive.Notably, Gln could affect the flavor of olive by regulating the activity of AS and sugar metabolism.
Using high performance liquid chromatography-tandem mass spectrometry(HPLC-MS/MS) combined with multivariate statistical analysis, fruits of the common olive ‘Changying' and the sweet olive ‘Qinglan 1 hao' were compared for differences in their free amino acid composition. Meanwhile, glutamine metabolism characteristics of the two olive varieties were analyzed. The aim was to provide the basis for understanding the contribution of amino acids to the differential flavor between common olive and sweet olive. By principal component analysis(PCA), partial least squares projection to latent structures discriminant analysis(PLS-DA) and significance analysis, the differential free amino acids between the two varieties were determined and correlation analysis was also conducted. The results showed that there was a difference in free amino acid composition between common olive and sweet olive with respect to the sweet amino acids serine(Ser)and glutamine(Gln) as well as umami glutamate(Glu). The three amino acids were all more abundant in ‘Qinglan 1 hao'and made greater contribution to the flavor of this variety. Their average contents were in the descending order: Glu > Gln >Ser for both varieties, and their differences between the two varieties were in descending order: Ser > Gln > Glu, overall suggesting that Gln was a more important amino acid. The Gln content of ‘Changying' was significantly negatively correlated with asparagine synthetase(AS) activity(P < 0.05) and soluble sugar with correlation coefficient(r) of-0.792 and-0.711,respectively, whereas no significant correlation with sucrose or reducing sugar was found(P > 0.05). The Gln content of‘Qinglan 1 hao' was not significantly correlated with AS activity, while it was highly significantly correlated with sucrose(P < 0.05, r = 0.953) and soluble sugar(P < 0.05, r = 0.791) and negatively correlated with reducing sugar(P < 0.05,r =-0.780). Hence, the correlations of Gln content with the metabolic enzyme activities and sugars differed between both varieties. In conclusion, free amino acids made different contributions to the flavor of common olive and sweet olive.Notably, Gln could affect the flavor of olive by regulating the activity of AS and sugar metabolism.
引文
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[23]WU Y,SHANG M.Characterization of principal compositions in the roots of Angelica sinensis by HPLC-ESI-MSn and chemical comparison of its different parts[J].Journal of Chinese Pharmaceutical Sciences,2014,23(6):393-402.DOI:10.5246/jcps.2014.06.053.
[24]杨文潮,杨洁,段金廒,等.南五味子中氨基酸类成分分析评价[J].中药材,2016,39(2):342-347.DOI:10.13863/j.issn1001-4454.2016.02.027.
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[26]侯娅,马阳,邹立思,等.基于UPLC-Trilpe TOF-MS/MS技术分析不同产地太子参的差异化学成分[J].质谱学报,2015,36(4):359-366.DOI:10.7538/zpxb.youxian.2015.0019.
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[30]蒋滢,徐颖,朱庚伯.人类味觉与氨基酸味道[J].氨基酸和生物资源,2002(4):70.
[31]武彦文,欧阳杰.氨基酸和肽在食品中的呈味作用[J].中国调味品,2001(1):21-24.DOI:10.3969/j.issn.1000-9973.2001.01.007.
[32]高肖飞.植物叶片中游离氨基酸的测定及其对大气氮沉降的响应[D].南昌:南昌大学,2015:77.
[33]李志霞,聂继云,闫震,等.响应面法对3,5-二硝基水杨酸比色法测定水果中还原糖含量条件的优化[J].分析测试学报,2016,35(10):1283-1288.DOI:10.3969/j.issn.1004-4957.2016.10.011.
[34]涂宇鹏.大肠杆菌BL21(DE3)氨基葡萄糖代谢调控关键酶的初步研究[D].上海:复旦大学,2011.
[35]李崇阳,李艳,牟德华.氨基葡萄糖硫酸软骨素促进骨骼健康作用研究进展[J].食品科学,2015,36(23):382-385.DOI:10.7506/spkx1002-6630-201523068.
[2]林玉芳.福建橄榄(Canarium album(Lour.)Raeusch)若干功能成分和品质相关指标的研究[D].福州:福建农林大学,2012:1-4.
[3]谢倩.橄榄(Canarium album(Lour.)Raeusch)果实发育成熟过程多酚及相关酶活性研究[D].福州:福建农林大学,2014:1-7.
[4]林聪.闽江流域橄榄(Canavium album Raeusch)生产与产业化对策研究[D].福州:福建农林大学,2011:1-5.
[5]彭勃,苗明三,朱平生,等.橄榄解酒饮对小鼠急性酒精性肝损伤胃肠组织病理学的影响[J].中医药学刊,2003,21(8):1231-1236.DOI:10.3969/j.issn.1673-7717.2003.08.005.
[6]彭勃,苗明三,王颖芳,等.橄榄解酒饮对大鼠急性酒精性肝损伤肝组织病理形态的影响[J].中国医药学报,2004,19(8):468-470.DOI:10.3969/j.issn.1673-1727.2004.08.006.
[7]王占恩,王燕青,李升刚,等.青果蠲毒散救治河豚鱼中毒临床观察[J].中国中西医结合杂志,2003,23(8):592.DOI:10.3321/j.issn:1003-5370.2003.08.037.
[8]杨桂林,何颖,董小娟,等.青果总黄酮的急性毒性及镇痛作用研究[J].安徽农业科学,2012,40(5):2674-2675.DOI:10.3969/j.issn.0517-6611.2012.05.044.
[9]王燕平,陈勤,周志钦,等.橄榄基因组AFLP扩增体系的优化[J].果树学报,2012,29(3):505-511.
[10]李泽坤.橄榄(Canarium album(Lour.)Raeusch)果实成熟发育蔗糖代谢变化研究[D].福州:福建农林大学,2016.
[11]池毓斌.橄榄果实品质特性及其代谢组学的初步研究[D].福州:福建农林大学,2017:100.
[12]王齐,朱伟伟,苏丹,等.蒲桃中氨基酸组成与含量对其营养与风味的影响[J].食品科学,2012,33(16):204-207.
[13]鲁敏,安华明,赵小红.无籽刺梨与刺梨果实中氨基酸分析[J].食品科学,2015,36(14):118-121.DOI:10.7506/spkx1002-6630-2015140023.
[14]PRIPIS-NICOLAU L,DE R G,BERTRAND A,et al.Formation of flavor components by the reaction of amino acid and carbonyl compounds in mild conditions[J].Journal of Agricultural and Food Chemistry,2000,48(9):3761-3766.DOI:10.1021/jf991024w.
[15]CHEN G,LI J,SUN Z,et al.Rapid and sensitive ultrasonic-assisted derivatisation microextraction(UDME)technique for bitter taste-free amino acids(FAA)study by HPLC-FLD[J].Food Chemistry,2014,143:97-105.DOI:10.1016/j.foodchem.2013.07.099.
[16]叶发荣,韩秀梅,肖钧,等.‘红阳’猕猴桃果实氨基酸含量及组成分析[J].中国园艺文摘,2015,28(5):1-3.
[17]万继锋,吴如健,韦晓霞,等.橄榄果实中糖和氨基酸组成与含量分析[J].福建农业学报,2013,28(5):472-477.DOI:10.3969/j.issn.1008-0384.2013.05.013.
[18]谢晓琼.橄榄(Canarium album(Lour.)Raeusch)果实若干生理生化指标的研究[D].福州:福建农林大学,2007:25-28.
[19]何志勇.橄榄酚类化合物的分离纯化和结构研究[D].无锡:江南大学,2007:22.
[20]车兰兰.液相色谱及液质联用法测定三种山西特色食品中游离氨基酸[D].太原:山西大学,2013.
[21]周文芳.血浆中氨基酸检测技术探讨[J].氨基酸和生物资源,2012,34(3):25-28.DOI:10.3969/j.issn.1006-8376.2012.03.007.
[22]马戎,鲍峰伟,冯文宁,等.液相色谱串联质谱法测定烟草中的游离氨基酸[J].中国测试,2013,39(2):34-37.
[23]WU Y,SHANG M.Characterization of principal compositions in the roots of Angelica sinensis by HPLC-ESI-MSn and chemical comparison of its different parts[J].Journal of Chinese Pharmaceutical Sciences,2014,23(6):393-402.DOI:10.5246/jcps.2014.06.053.
[24]杨文潮,杨洁,段金廒,等.南五味子中氨基酸类成分分析评价[J].中药材,2016,39(2):342-347.DOI:10.13863/j.issn1001-4454.2016.02.027.
[25]王学奎.植物生理生化实验原理与技术[M].2版.北京:高等教育出版社,2006.
[26]侯娅,马阳,邹立思,等.基于UPLC-Trilpe TOF-MS/MS技术分析不同产地太子参的差异化学成分[J].质谱学报,2015,36(4):359-366.DOI:10.7538/zpxb.youxian.2015.0019.
[27]吴宏伟,李洪梅,唐力英,等.代谢组学方法研究姜黄根茎及块根次生代谢产物表达差异[J].分析化学,2012,40(5):713-717.DOI:10.3724/SP.J.1096.2012.11025.
[28]TRYGG J,HOLMES E,LUNDSTEDT T.Chemometrics in metabonomics[J].Journal of Proteome Research,2007,6(2):469-479.DOI:10.1021/pr060594q.
[29]余欣尉,吴谦,吕望,等.基于液相色谱-质谱联用技术的肺癌细胞代谢组学分析[J].色谱,2013,31(7):691-696.DOI:10.3724/SP.J.1123.2012.12039.
[30]蒋滢,徐颖,朱庚伯.人类味觉与氨基酸味道[J].氨基酸和生物资源,2002(4):70.
[31]武彦文,欧阳杰.氨基酸和肽在食品中的呈味作用[J].中国调味品,2001(1):21-24.DOI:10.3969/j.issn.1000-9973.2001.01.007.
[32]高肖飞.植物叶片中游离氨基酸的测定及其对大气氮沉降的响应[D].南昌:南昌大学,2015:77.
[33]李志霞,聂继云,闫震,等.响应面法对3,5-二硝基水杨酸比色法测定水果中还原糖含量条件的优化[J].分析测试学报,2016,35(10):1283-1288.DOI:10.3969/j.issn.1004-4957.2016.10.011.
[34]涂宇鹏.大肠杆菌BL21(DE3)氨基葡萄糖代谢调控关键酶的初步研究[D].上海:复旦大学,2011.
[35]李崇阳,李艳,牟德华.氨基葡萄糖硫酸软骨素促进骨骼健康作用研究进展[J].食品科学,2015,36(23):382-385.DOI:10.7506/spkx1002-6630-201523068.