利用10种食品级蛋白酶分别对牛心肌进行酶解处理,并探究酶解产物的呈味特性与肽结构之间的内在联系。分别采用Protease A、Protease P、Proteax、Flavourzyme、Alcalase、Papain、Bromelain、Protamex、Netutrase和Sumizyme BNP-L对牛心肌进行酶解,分析蛋白酶解物的水解度、分子质量分布、酶特异性,肽谱图与呈味特性的关系。研究结果表明,Protease A作为一种内外肽酶的复合酶制剂,是制备高鲜味、低苦味酶解物的最佳蛋白酶;然而,2种内肽酶(木瓜蛋白酶和菠萝蛋白酶)会导致蛋白酶解物产生苦味。偏最小二乘回归(partial least squares regression,PLSR)分析表明,酶解物的鲜味与低分子质量肽(<1 kDa)具有一定的正相关性,而其苦味与分子质量为0.5~1 kDa的肽组分呈正相关。蛋白酶特异性和肽谱图分析揭示,由于10种蛋白酶具有不同的酶切位点,导致相应的蛋白酶解物形成了不同的肽谱图,从而阐明蛋白酶解物的不同呈味特性。通过内肽酶和外肽酶联合酶解作用牛心肌,可以有效地减少蛋白酶解物的苦味并增加其鲜味。
Bovine cardiac muscle was hydrolyzed by ten food-grade proteases, namely Protease A, Protease P, ProteAX, Flavourzyme, Alcalase, Papain, Bromelain, Protamex, Netutrase and Sumizyme BNP-L, and the relationship between taste characteristics and peptide structure of the protein hydrolysates was investigated. The degree of hydrolysis, molecular weight distribution, enzyme specificity and peptide profile of protein hydrolysates were analyzed. Results showed that Protease A, a compound enzyme preparation with exopeptidases and endopeptidases, was the optimal protease for preparing low-bitterness and high-umami protein hydrolysates. However, two endopeptidases (Papain and Bromelain) elicited a bitter taste in the protein hydrolysates. Partial least squares regression (PLSR) analysis showed that the umami taste of protein hydrolysates was positively correlated with low molecular weight (< 1 kDa) peptides, while the bitterness of protein hydrolysates was positively correlated with peptides with molecular weight of 0.5-1 kDa. The analysis of protease specificity and peptide profile revealed that ten proteases had different enzymatic cleavage specificity, which led to the formation of different peptide profiles, thus elucidating the different taste characteristics of protein hydrolysates. Endopeptidase combined with exopeptidase hydrolysis of bovine cardiac muscle can effectively reduce the bitterness and enhance the umami of protein hydrolysates.
[1] HENCHION M, HAYES M, MULLEN A M, et al.Future protein supply and demand:Strategies and factors influencing a sustainable equilibrium[J].Foods, 2017, 6(7):53.
[2] TOLDRÁ F, MORA L, REIG M.New insights into meat by-product utilization[J].Meat Science, 2016, 120:54-59.
[3] LONG J M, MOHAN A.Development of meat powder from beef byproduct as value-added food ingredient[J].LWT, 2021, 146:111460.
[4] MULLEN A M, ÁLVAREZ C, ZEUGOLIS D I, et al.Alternative uses for co-products:Harnessing the potential of valuable compounds from meat processing chains[J].Meat Science, 2017, 132:90-98.
[5] FU Y, CHEN J R, BAK K H, et al.Valorisation of protein hydrolysates from animal by-products:Perspectives on bitter taste and debittering methods:A review[J].International Journal of Food Science & Technology, 2019, 54(4):978-986.
[6] MAEHASHI K, HUANG L.Bitter peptides and bitter taste receptors[J].Cellular and Molecular Life Sciences, 2009, 66(10):1 661-1 671.
[7] FU Y, BAK K H, LIU J, et al.Protein hydrolysates of porcine hemoglobin and blood:Peptide characteristics in relation to taste attributes and formation of volatile compounds[J].Food Research International, 2019, 121:28-38.
[8] 王知非, 林璐, 孙伟峰, 等.苦味肽和苦味受体研究进展[J].中国调味品, 2016, 41(9):152-156.
WANG Z F, LIN L, SUN W F, et al.Research progress of bitter peptides and bitter receptors[J].China Condiment, 2016, 41(9):152-156.
[9] NEWMAN J, EGAN T, HARBOURNE N, et al.Correlation of sensory bitterness in dairy protein hydrolysates:Comparison of prediction models built using sensory, chromatographic and electronic tongue data[J].Talanta, 2014, 126:46-53.
[10] SPELLMAN D, MCEVOY E, O′CUINN G, et al.Proteinase and exopeptidase hydrolysis of whey protein:Comparison of the TNBS, OPA and pH stat methods for quantification of degree of hydrolysis[J].International Dairy Journal, 2003, 13(6):447-453.
[11] LIU B Y, ZHU K X, GUO X N, et al.Changes in the enzyme-induced release of bitter peptides from wheat gluten hydrolysates[J].RSC Advances, 2016, 6(104):102 249-102 257.
[12] FU Y, LIU J, ZHANG W, et al.Exopeptidase treatment combined with Maillard reaction modification of protein hydrolysates derived from porcine muscle and plasma:Structure-taste relationship[J].Food Chemistry, 2020, 306:125613.
[13] CHEUNG L K Y, ALUKO R E, CLIFF M A, et al.Effects of exopeptidase treatment on antihypertensive activity and taste attributes of enzymatic whey protein hydrolysates[J].Journal of Functional Foods, 2015, 13:262-275.
[14] ZHANG J N, SUN-WATERHOUSE D, SU G W, et al.New insight into umami receptor, umami/umami-enhancing peptides and their derivatives:A review[J].Trends in Food Science & Technology, 2019, 88:429-438.
[15] LIU Z Y, ZHU Y W, WANG W L, et al.Seven novel umami peptides from Takifugu rubripes and their taste characteristics[J].Food Chemistry, 2020, 330:127204.
[16] WANG W L, ZHOU X R, LIU Y.Characterization and evaluation of umami taste:A review[J].TrAC Trends in Analytical Chemistry, 2020, 127:115876.
[17] KIM M J, SON H J, KIM Y, et al.Umami-bitter interactions:The suppression of bitterness by umami peptides via human bitter taste receptor[J].Biochemical and Biophysical Research Communications, 2015, 456(2):586-590.
[18] 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.
[19] TEMUSSI P A.The good taste of peptides[J].Journal of Peptide Science, 2012, 18(2):73-82.
[20] 姜莉莉, 李杏元.呈味肽应用于食品的研究进展[J].中国调味品, 2019,44(11):187-189.
JIANG L L, LI X Y.Research progress on application of taste peptides in food[J].China Condiment, 2019, 44(11):187-189.
[21] ZHU X P, SUN-WATERHOUSE D, CHEN J H, et al.Bitter-tasting hydrophobic peptides prepared from soy sauce using aqueous ethanol solutions influence taste sensation[J].International Journal of Food Science & Technology, 2020, 55(1):146-156.
[22] ASPEVIK T, TOTLAND C, LEA P, et al.Sensory and surface-active properties of protein hydrolysates based on Atlantic salmon (Salmo salar) by-products[J].Process Biochemistry, 2016, 51(8):1 006-1 014.
[23] SCHINDLER A, DUNKEL A, STAÄHLER F, et al.Discovery of salt taste enhancing arginyl dipeptides in protein digests and fermented fish sauces by means of a sensomics approach[J].Journal of Agricultural and Food Chemistry, 2011, 59(23):12 578-12 588.
