Isolation, Purification and Structure Identification of Umami Peptide from Air-dried and Fermented Megalobrama Amblycephala

Abstract

Abstract: In this paper, umami peptides were isolated and purified from air-dried and fermented Megalobrama Amblycephala using ultrafiltration, nanofiltration and reversed phase high performance liquid chromatography (RP-HPLC) and the umami was measured by using electronic tongue. Matrix assisted laser desorption ionization time of flight mass spectrometer (MALDI-TOF-MS), automatic amino acid analyzer and fourier transform infrared spectroscopy (FTIR) was used to measure the molecular mass, amino acid composition and secondary structure of umami peptide to investigate the relationship between structure and umami of peptide. The results showed that a single umami peptide P2a-2 with a molecular mass of 1304.59 Da was purified and identified from air-dried and fermented Megalobrama Amblycephala. Moreover, the umami amino acids such as glutamic acid (21.97%), alanine (10.67%) and glycine (9.51%), the umami enhancer histidine (27.54%), and the secondary structure (beta- turn) with 90.16%, which were found in P2a-2, suggesting that P2a-2 presented the heavy umami due to the high content of umami amino acid and the secondary structure with beta-turn.

Key words: air-dried and fermented Megalobrama Amblycephala, peptide, umami, purification, secondary structure

CLC Number:

TS254.4

References

[1] 赵俊仁, 程水明, 洪玮娣, 等. 风干武昌鱼中乳酸菌的分离鉴定及发酵性能研究[J]. 现代食品科技, 2014, 30(8): 100-105. DOI：10.13982/j.mfst.1673-9078.2014.08.022
[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(1): 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(1): 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(1): 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. 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, Food Chemistry, 2018, Available online. 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, R C GUO, et al. Taste, umami-enhance effect and amino acid sequence of peptides separated from silkworm pupa hydrolysate[J]. Food Research International, 2018, 108(1): 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(1): 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(1): 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/spkx 1002- 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.
[22] 余瑞元，袁明秀，陈丽蓉，等．生物化学实验原理和方法（第二版）[M]．北京：北京大学出版社，2005: 46-153.
[23] HASNI I, BOURASSA P, HAMDANI S, et al. Interaction of milk alpha- and beta-caseins with tea polyphenols [J]. Food Chemistry, 2011, 126(2): 630-639. DOI: 10.1016/j.foodchem.2010.11.087
[24] FENNEMA O R 著, 王璋, 许时婴, 江波, 等译. 食品化学（第三版）[M]. 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(1): 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(1): 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(1): 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（第三版）[M]. 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