风干武昌鱼中鲜味肽的分离纯化及二级结构分析
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摘要
采用电子舌测定鲜味,超纳滤、反相高效液相色谱从风干武昌鱼中分离纯化出纯度均一的鲜味肽,基质辅助激光解析电离-飞行时间质谱仪、氨基酸自动分析仪、傅里叶变换红外光谱测定鲜味肽的分子质量、氨基酸组成和二级结构,探讨鲜味肽结构与风味的关系。结果显示:从风干武昌鱼中纯化得到了一种鲜味较强的肽P2a-2,其分子质量为1 304.59 Da。P2a-2的鲜味氨基酸谷氨酸(21.97%)、甘氨酸(10.67%)和丙氨酸(9.51%)比例较高,且助味氨基酸组氨酸的比例最高(27.54%),β-转角为主要的二级结构(90.16%),推断P2a-2呈现较佳鲜味与其高含量的鲜味氨基酸和β-转角相关。
In this paper, a umami peptide was isolated and purified from air-dried and fermented Megalobrama amblycephala using ultrafiltration, nanofiltration and reversed phase high performance liquid chromatography(RP-HPLC).The umami taste was measured with an electronic tongue. Matrix assisted laser desorption ionization time of flight mass spectrometry(MALDI-TOF-MS), an automatic amino acid analyzer and Fourier transform infrared spectroscopy(FTIR)were used to measure the molecular mass, amino acid composition and secondary structure of the umami peptide and furthermore, we investigated the relationship between structure and umami taste. The results showed that a single umami peptide P2a-2 with a molecular mass of 1 304.59 Da was obtained and identified. Moreover, the umami amino acids glutamic acid(21.97%), alanine(10.67%) and glycine(9.51%), as well as the umami enhancer histidine(27.54%) were found in P2a-2, and the secondary structure was beta-turn(90.16%). We deduced that P2a-2's strong umami taste may be due to the high content of umami amino acid and beta-turn structure.
In this paper, a umami peptide was isolated and purified from air-dried and fermented Megalobrama amblycephala using ultrafiltration, nanofiltration and reversed phase high performance liquid chromatography(RP-HPLC).The umami taste was measured with an electronic tongue. Matrix assisted laser desorption ionization time of flight mass spectrometry(MALDI-TOF-MS), an automatic amino acid analyzer and Fourier transform infrared spectroscopy(FTIR)were used to measure the molecular mass, amino acid composition and secondary structure of the umami peptide and furthermore, we investigated the relationship between structure and umami taste. The results showed that a single umami peptide P2a-2 with a molecular mass of 1 304.59 Da was obtained and identified. Moreover, the umami amino acids glutamic acid(21.97%), alanine(10.67%) and glycine(9.51%), as well as the umami enhancer histidine(27.54%) were found in P2a-2, and the secondary structure was beta-turn(90.16%). We deduced that P2a-2's strong umami taste may be due to the high content of umami amino acid and beta-turn structure.
引文
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[33]FUKE S,UEDA Y. Interaction between umami and other flavor characteristics[J]. Trends in Food Science and Technology,1996,7(12):407-411.DOI:10.1016/S0924-2244(96)10042-X.
[34]KONG J,YU S. Fourier transform infrared spectroscopic analysis of protein secondary structures[J]. Acta Biochimica et Biophysica Sinica,2007,39(8):549-559.DOI:10.1111/j.1745-7270.2007.00320.x.
[2]HESSELTINE C W,WANG H L. Traditional fermented foods[J]. Biotechnology and Bioengineering,1967,9(3): 275-288. DOI:10.1002/bit.260090302.
[3]FU Y,LIU J,HANSEN E T,et al. Structural characteristics of low bitter and high umami protein hydrolysates prepared from bovine muscle and porcine plasma[J]. Food Chemistry,2018,257: 163-171. DOI:10.1016/ j.foodchem.2018.02.159.
[4]KIM Y,KIM E Y,SON H J,et al. Identification of a key umami-active fraction in modernized Korean soy sauce and the impact thereof on bitter-masking[J]. Food Chemistry,2017,233: 256-262. DOI:10.1016/j.foodchem.2017.04.123.
[5]KONG Y,YANG X,DING Q,et al. Comparison of non-volatile umami components in chicken soup and chicken enzymatic hydrolysate[J]. Food Research International,2017,102:559-566.DOI:10.1016/j.foodres.2017.09.038.
[6]ZHANG Y,VENKITASAMY C,PAN Z,et al. Novel umami ingredients: umami peptides and their taste[J]. Journal of Food Science,2017,82(1):16-23.DOI:10.1111/1750-3841.13576.
[7]DANG Y L,GAO X C,MA F M,et al. Comparison of umami taste peptides in water-soluble extractions of Jinhua and Parma hams[J]. LWT-Food Science and Technology,2015,60(2): 1179-1186. DOI:10.1016/j.lwt.2014.09.014.
[8]ISTIQAMAH A,LIOE H N,ADAWIYAH D R. Umami compounds present in low molecular umami fractions of Asam Sunti: a fermented fruit of Averrhoa bilimbi L[J]. Food Chemistry,2019,270: 338-343. DOI:10.1016/j.foodchem.2018.06.131.
[9]RHYU M R,KIM E Y. Umami taste characteristics of water extract of doenjang,a Korean soybean paste: low-molecular acidic peptides may be a possible clue to the taste[J]. Food Chemistry,2011,127(3): 1210-1215.DOI:10.1016/j.foodchem.2011.01.128.
[10]SHAH A K M A,OGASAWARA M,MAKOTO E,et al. Identification and sensory evaluation of flavour enhancers in Japanese traditional dried herring fillet[J]. Food Chemistry,2010,122(1): 249-253. DOI:10.1016/j.foodchem.2010.02.072.
[11]SU G W,CUI CHUN,ZHENG L,et al. Isolation and identification of two novel umami and umami-enhancing peptides from peanut hydrolysate by consecutive chromatography and MALDI-TOF/TOF MS[J]. Food Chemistry,2012,135(2): 479-485. DOI:10.1016/j.foodchem.2012.04.130.
[12]YANG H Y,YANG S N,KONG J L,et al. Obtaining information about protein secondary structures in aqueous solution using Fourier transform IR spectroscopy[J]. Nature Protocols,2015,10(3): 382-396. DOI:10.1038/nprot.2015.024.
[13]YU Z L,JIANG H R,GUO R C,et al. Taste,umami-enhance effect and amino acid sequence of peptides separated from silkworm pupa hydrolysate[J].Food Research International,2018,108:144-150.DOI:10.1016/j.foodres.2018.02.047.
[14]ZHANG M X,WANG X C,LIU Y,et al. Isolation and identification of flavour peptides from Puffer fish (Takifugu obscurus) muscle using an electronic tongue and MALDI-TOF/TOF MS/MS[J]. Food Chemistry,2012,135:1463-1470.DOI:10.1016/j.foodchem. 2012. 06.026.
[15]ZHUANG M Z,LIN L Z,ZHAO M M,et al.Sequence,taste and umami-enhancing effect of the peptides separated from soy sauce[J]. Food Chemistry,2016,206(1): 174-181. DOI:10.1016/j.foodchem.2016.03.058.
[16]BAGNASCO L,COSULICH M E,SPERANZA G,et al.Application of a voltammetric electronic tongue and near infrared spectroscopy for a rapid umami taste assessment[J]. Food Chemistry,2014,157(8): 421-428.DOI:10.1016/j.foodchem.2014.02.044.
[17]VLASOV Y,LEGIN A,RUDNITSKAYA A. Electronic tongue and their analytical application[J]. Analytical and Bioanalytical Chemistry,2002,373:136-146.DOI:10.1007/s00216-002-1310-2.
[18]刘静泊,陈季旺,夏文水,等.风干武昌鱼的营养及挥发性成分[J].食品科学,2015,36(18):80-84.DOI:10.7506/spkx1002-6630-201518014.
[19]国家卫生和计划生育委员会.食品中水分的测定:GB 5009.3-2016[S].北京:中国标准出版社,2016:1-2.
[20]国家卫生和计划生育委员会.食品中灰分的测定:GB 5009.4-2016[S].北京:中国标准出版社,2016:1-3.
[21]国家食品药品监督管理总局.食品中蛋白质的测定:GB 5009.5-2016[S].北京:中国标准出版社,2016:1-2.
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[23]HASNI I,BOURASSA P,HAMDANI S,et al. Interaction of milk α-andβ-caseins with tea polyphenols[J]. Food Chemistry,2011,126(2):630-639.DOI:10.1016/j.foodchem.2010.11.087.
[24]FENNEMA O R.食品化学[M].3版.王璋,许时婴,江波,等译.北京:中国轻工业出版社,2003:53-231.
[25]INOUE Y,KATO S,SAIKUSA M,et al. Analysis of the cooked aroma and odorants that contribute to umami aftertaste of soy miso (Japanese soybean paste)[J]. Food Chemistry,2016,213: 521-528. DOI:10.1016/j.foodchem.2016.06.106.
[26]ZHANG L Y,PETERSON D G. Identification of a novel umami compound in potatoes and potato chips[J]. Food Chemistry,2018,240: 1219-1226.DOI:10.1016/ j.food chem.2017.08.043.
[27]ISHIWATARI N,FUKUOKA M,HAMADA-SATO N,et al.Decomposition kinetics of umami component during meat cooking[J]. Journal of Food Engineering,2013,119(2): 324-331. DOI:10.1016/j.jfoodeng.2013.05.040.
[28]LIOEA H N,KINJOA A,YASUDA S,et al. Taste and chemical characteristics of low molecular weight fractions from tofuyo-Japanese fermented soybean curd[J]. Food Chemistry,2018,252: 265-270. DOI:10.1016/j.foodchem.2018.01.117.
[29]ClAEYS E,DE SMET S,BALCAEN A,et al. Quantification of fresh meat peptides by SDS-PAGE in relation to ageing time and taste intensity[J].Meat Science,2004,67(2):281-288.DOI:10.1016/j.meatsci.2003.11.001.
[30]王竟岩,朱圣庚,徐长法.生物化学[M].3版.北京:高等教育出版社,2002:105-201.
[31]NISHIMURA T,KATO H. Taste of free amino acids and peptides[J]. Food Review International,1988,4(2): 175-194. DOI:10.1080/87559128809540828.
[32]SENTANDEU M A,STOEVA S,ARISTOY M C,et al. Identification of small peptides generated in Spanish dry-cured ham[J]. Journal of Food Science,2003,68(1): 64-69. DOI:10.1111/j.1365-2621.2003.tb14115.x.
[33]FUKE S,UEDA Y. Interaction between umami and other flavor characteristics[J]. Trends in Food Science and Technology,1996,7(12):407-411.DOI:10.1016/S0924-2244(96)10042-X.
[34]KONG J,YU S. Fourier transform infrared spectroscopic analysis of protein secondary structures[J]. Acta Biochimica et Biophysica Sinica,2007,39(8):549-559.DOI:10.1111/j.1745-7270.2007.00320.x.