醋蛋液水解物的类蛋白反应修饰及其对胆酸结合能力的影响
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摘要
[目的]探讨类蛋白反应在醋蛋液水解过程中对其胆酸结合能力的影响。[方法]采用胃蛋白酶水解醋蛋液,制备水解度为43. 61%,胆酸结合能力为64.01%的醋蛋液水解物,并以游离氨基减少量为试验指标,利用响应面法优化胃蛋白酶催化的类蛋白反应修饰条件,得到不同修饰程度的产物,测定其结合胆酸的能力。[结果]修饰产物的胆酸结合能力均提高,并且结合能力最高的修饰产物的结合能力达到102.1%,该修饰产物通过乙醇∶水(7∶3)或乙醇∶水(3∶7)比例的溶剂进行离心分级后,上清液(沉淀)部分的胆酸结合能力高于(低于)修饰产物,并通过对该修饰产物和上清液部分、沉淀部分的进一步酶水解处理的结果显示,酶水解会导致它们的结合能力降低,但是仍然高于最初的醋蛋液水解物。[结论]类蛋白反应可以提高醋蛋液水解物的胆酸结合能力和对蛋白酶抵抗的能力。
[Objective]The impact of solvent fractionation and protease digestion on the bile acid-binding capacity of plastein reaction-stressed vinegar-egg hydrolysates was discussed.[Method]Vinegar-egg was hydrolyzed by pepsin to obtain a hydrolysate with a degree of hydrolysis of43.61% and a bile acid-binding capacity of 64.01%.The decrease of free amino groups of the modified hydrolysate which was as the response was measured to obtain appropriate reaction condition.[Result] Bile acid-binding capacity of modified hydrolysate was improved,the treated hydrolysates were obtained and exhibited the highest bile acid-binding capacity of 102.1%.This modified hydrolysate was separated by applying ethanol-water of 7 ∶3 or 3 ∶7,the obtained supernatant( precipitate) fractionates have higher( lower) capacity to bile acid than the parent substrate.The further enzymatic hydrolysis of the supernatant and precipitation fractions could result in the impairment of bile acid-binding capacity.[Conclusion]Plastein reaction could enhance bile acid-binding capacity and protease resistance of vinegar-egg hydrolysate.
[Objective]The impact of solvent fractionation and protease digestion on the bile acid-binding capacity of plastein reaction-stressed vinegar-egg hydrolysates was discussed.[Method]Vinegar-egg was hydrolyzed by pepsin to obtain a hydrolysate with a degree of hydrolysis of43.61% and a bile acid-binding capacity of 64.01%.The decrease of free amino groups of the modified hydrolysate which was as the response was measured to obtain appropriate reaction condition.[Result] Bile acid-binding capacity of modified hydrolysate was improved,the treated hydrolysates were obtained and exhibited the highest bile acid-binding capacity of 102.1%.This modified hydrolysate was separated by applying ethanol-water of 7 ∶3 or 3 ∶7,the obtained supernatant( precipitate) fractionates have higher( lower) capacity to bile acid than the parent substrate.The further enzymatic hydrolysis of the supernatant and precipitation fractions could result in the impairment of bile acid-binding capacity.[Conclusion]Plastein reaction could enhance bile acid-binding capacity and protease resistance of vinegar-egg hydrolysate.
引文
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[19]SUN H,LI T J,ZHAO X H.Ace Inhibition and enzymatic resistance in vitro of a casein hydrolysate subjected to plastein reaction in the presence of extrinsic proline and ethanol-or methanol-water fractionation[J].International journal of food properties,2014,17(2):386-398.
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[21]WEI X,LI T J,ZHAO X H.Coupled neutrase-catalyzed plastein reaction mediated the ACE-Inhibitory activity in vitro of casein hydrolysates prepared by alcalase[J]. International journal of food properties,2013,16(2):429-443.
[22]LIN Y H,TSAI J S,CHEN G W.Purification and identification of hypocholesterolemic peptides from freshwater clam hydrolysate with in vitro gastrointestinal digestion[J].Journal of food biochemistry,2017,41(4):12385.
[23]UDENIGWE C C,WU S H,DRUMMOND K,et al.Revisiting the prospects of plastein:Thermal and simulated gastric stability in relation to the antioxidative capacity of casein plastein[J].Journal of agricultural and food chemistry,2014,62(1):130-135.
[24]HIGAKI N,SATO K,SUDA H,et al.Evidence for the existence of a soybean resistant protein that captures bile acid and stimulates its fecal excretion[J].Journal of the agricultural chemical society of Japan,2006,70(12):2844-2852.
[25]BROWNSELL V L,WILLIAMS R J H,ANDREWS A T.Application of the plastein reaction to mycoprotein:II.Plastein properties[J].Food chemistry,2001,72(3):337-346.
[2]NAGAOKA S,FUTAMURA Y,MIWA K,et al.Identification of novel hypocholesterolemic peptides derived from bovine milk beta-lactoglobulin[J].Biochemical and biophysical research communications,2001,281(1):11-17.
[3]SHIRATA S,ODA K,ONODERA-MASUOKA N,et al.Hypocholesterolemic effect of indigestible fraction of Chlorella regularis in cholesterol-fed rats[J].Journal of nutritional science and vitaminology,2001,47(6):373-377.
[4]KONGO-DIA-MOUKALA J U,ZHANG H,IRAKOZE P C.In vitro binding capacity of bile acids by defatted corn protein hydrolysate[J].International journal of molecular sciences,2011,12(2):1066-1080.
[6]MULLALLY M M,MEISEL H,FITZGERALD R J.Angiotensin-I-converting enzyme inhibitory activities of gastric and pancreatic proteinase digests of whey proteins[J].International dairy journal,1997,7(5):299-303.
[6]PIHLANTO-LEPPA LA A,ROKKA T,KORHONEN H.Angiotensin I converting enzyme inhibitory peptides derived from bovine milk proteins[J].International dairy journal,1998,8(4):325-331.
[7]杨锋,陈锦屏,吴莉莉.醋蛋多肽血管紧张素转化酶抑制活性的稳定性研究[J].食品工业科技,2012,33(11):151-153,156.
[8]杨锋,陈锦屏,林心怡.美拉德反应对醋蛋多肽抗氧化活性的影响[J].食品科学,2013,34(3):137-140.
[9]STEVENSON D E,OFMAN D J,MORGAN K R,et al.Protease-catalyzed condensation of peptides as a potential means to reduce the bitter taste of hydrophobic peptides found in protein hydrolysates[J].Enzymeµbial technology,1998,22(2):100-110.
[10] ASHLEY D V,TEMLER R,BARCLAY D,et al.Amino acid-enriched plasteins:A source of limiting amino acids for the weanling rat[J].Journal of nutrition,1983,113(1):21-27.
[11]YAMASHITA M,ARAI S,AMANO Y,et al.A novel one-step process for enzymatic incorporation of amino acids into proteins:Application to soy protein and flour for enhancing their methionine levels[J]. Agricultural and biological chemistry,1979,43(5):1065-1068.
[12]QIAN F,WANG Y,WEN Z J,et al.Plastein reaction enhanced bile-acid binding capacity of soybean protein hydrolysates and whey protein hydrolysates[J].Journal of food science&technology,2018,55(3):1021-1027.
[13]HOWARD A,UDENIGWE C C.Mechanisms and prospects of food protein hydrolysates and peptide-induced hypolipidaemia[J]. Food&function,2013,4(1):41-50.
[14]UDENIGWE C C,MOHAN A,WU S H.Peptide aggregation during plastein reaction enhanced bile acid-binding capacity of enzymatic chicken meat hydrolysates[J].Journal of food biochemistry,2015,39(3):344-348.
[15]CHURCH F C,SWAISGOOD H E,PORTER D H,et al.Spectrophotometric assay using o-phthaldialdehyde for determination of proteolysis in milk and isolated milk proteins[J].Journal of dairy science,1983,66(6):1219-1227.
[16]SPELLMAN D,MCEVOY E,O’CUINN G,et al.Proteinase and exopeptidase hydrolysis of whey protein:Comparison of the TNBS,OPA and p H stat methods for quantification of degree of hydrolysis[J]. International dairy journal,2003,13(6):447-453.
[17]ADLER-NISSEN J.Determination of the degree of hydrolysis of food protein hydrolysates by trinitrobenzenesulfonic acid[J].Journal of agricultural and food chemistry,1979,27(6):1256-1262.
[18] SAIGA A,TANABE S,NISHIMURA T.Antioxidant activity of peptides obtained from porcine myofibrillar proteins by protease treatment[J].Journal of agricultural and food chemistry,2003,51(12):3661-3667.
[19]SUN H,LI T J,ZHAO X H.Ace Inhibition and enzymatic resistance in vitro of a casein hydrolysate subjected to plastein reaction in the presence of extrinsic proline and ethanol-or methanol-water fractionation[J].International journal of food properties,2014,17(2):386-398.
[20]ZHANG Y,ZHAO X H.In vitro angiotensin I-converting enzyme inhibition of casein hydrolysate responsible for plastein reaction in ethanol-water medium,solvent fractionation,and protease digestion[J].International journal of food properties,2014,17(7):1577-1590.
[21]WEI X,LI T J,ZHAO X H.Coupled neutrase-catalyzed plastein reaction mediated the ACE-Inhibitory activity in vitro of casein hydrolysates prepared by alcalase[J]. International journal of food properties,2013,16(2):429-443.
[22]LIN Y H,TSAI J S,CHEN G W.Purification and identification of hypocholesterolemic peptides from freshwater clam hydrolysate with in vitro gastrointestinal digestion[J].Journal of food biochemistry,2017,41(4):12385.
[23]UDENIGWE C C,WU S H,DRUMMOND K,et al.Revisiting the prospects of plastein:Thermal and simulated gastric stability in relation to the antioxidative capacity of casein plastein[J].Journal of agricultural and food chemistry,2014,62(1):130-135.
[24]HIGAKI N,SATO K,SUDA H,et al.Evidence for the existence of a soybean resistant protein that captures bile acid and stimulates its fecal excretion[J].Journal of the agricultural chemical society of Japan,2006,70(12):2844-2852.
[25]BROWNSELL V L,WILLIAMS R J H,ANDREWS A T.Application of the plastein reaction to mycoprotein:II.Plastein properties[J].Food chemistry,2001,72(3):337-346.