羊乳清蛋白部分和深度水解工艺、水解物致敏性及肽谱分析
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(1.天津科技大学 食品科学与工程学院/食品营养与安全国家重点实验室, 天津 300457;2.宜品乳业(青岛)集团有限公司, 山东 青岛 266600)

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Partial and Extensive Hydrolysis Process, Hydrolysate Allergenicity and Peptide Profile of Goat Whey Protein
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(1. College of Food and Engineering/State Key Laboratory of Food Nutrition and Safety, Tianjin University of Science and Technology, Tianjin 300457, China;2. Yipin Dairy (Qingdao) Group Co. Ltd., Qingdao 266600, China)

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    摘要:

    利用羊乳清蛋白制备低致敏性配料是目前乳品工业的研究热点。乳清蛋白是乳中主要的蛋白质之一,也是引起婴幼儿过敏反应的主要成分,将蛋白质水解为小分子肽是降低其致敏性的有效方法。以山羊乳清蛋白为原料,研究了部分水解乳清蛋白和深度水解乳清蛋白的水解工艺和水解物特性(水解度、分子质量分布和β-乳球蛋白抗原性),并利用液相色谱串联质谱法(LC-MS/MS) 比较了部分水解和深度水解工艺中过敏表位酶切位点的差异。研究结果表明,中性蛋白酶和碱性蛋白酶对羊乳清蛋白水解效果较好,其中碱性蛋白酶的水解度最高,达21.26%。经电泳分析,单酶水解后的产物中仍存在大分子多肽链,深度水解工艺需要复合酶水解。在酶底比为4000U/g时,使用碱性蛋白酶在pH值为10.0、温度为55℃条件下水解羊乳清蛋白1.0h,部分水解产物的水解度为12.31%,分子质量在5kDa以下的多肽占95.18%,β-乳球蛋白抗原性下降率为9.40%。中性蛋白酶和碱性蛋白酶的质量比为1∶1,酶底比为6000U/g,在pH值为8.5、温度为50℃条件下,水解羊乳清蛋白3.0h,深度水解产物的水解度为35.58%,分子质量低于3kDa的多肽为97.26%,β-乳球蛋白抗原性下降率为40.97%。部分水解和深度水解均能破坏β-乳球蛋白的大部分过敏表位,但相较于部分水解,深度水解能更大程度地降低乳清蛋白的致敏性。研究旨在为低致敏性羊水解乳清蛋白的生产提供一定的理论参考。

    Abstract:

    The preparation of hypoallergenic ingredients using goat whey protein is a hot research topic in dairy industry. Whey protein is one of the main proteins in milk, and it is also the main substance that causes allergic reactions in infants and young children. The most common way to reduce protein sensitization is to hydrolyze protein into small molecular peptides. The hydrolysis process of partial and extensive hydrolysis, and the characteristics of hydrolysate (degree of hydrolysis, molecular weight distribution and β-Lg antigenicity) were studied with goat whey protein as raw materials. LC-MS/MS was used to compare the cleavage sites of allergic epitopes by partial hydrolysis and extensive hydrolysis. The results showed that neutral protease and alkaline protease hydrolysis had better hydrolysis effect on goat whey protein, in which alkaline protease had the highest hydrolysis degree of 21.26%. Analyzed by electropherograms, macromolecular polypeptide chains still existed in the products after single enzyme hydrolysis, and complex enzyme hydrolysis was required in the extensive hydrolysis process. At the enzyme to substrate ratio of 4000U/g, alkaline protease was used for hydrolysis at pH 10.0 and 55℃ for 1.0h. The degree of hydrolysis of partial hydrolysates was 12.31%, the molecular weight of polypeptides below 5kDa accounted for 95.18%, and the antigenicity of β-lactoglobulin decreased by 9.40%. When the mass ratio of neutral protease and alkaline protease was 1∶1, the enzyme to substrate ratio was 6000U/g, and the hydrolysis was conducted at pH 8.5 and 50℃ for 3.0h, the hydrolysis degree of the extensive hydrolysate was 35.58%, peptides with molecular weight less than 3kDa accounted for 97.26%, and the antigenicity of β-lactoglobulin decreased by 40.97%. Both partial and extensive hydrolysis could destroy most of the allergic epitopes of β-lactoglobulin, but extensive hydrolysis could reduce the allergenicity of whey protein to a greater extent than partial hydrolysis. The purpose of this study was to provide some reference for the production of hydrolyzed whey protein in hypoallergenic goat.

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李红娟,李梦凡,曹洪宇,刘雷,牟善波,于景华.羊乳清蛋白部分和深度水解工艺、水解物致敏性及肽谱分析[J].食品科学技术学报,2024,42(3):81-91.

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  • 在线发布日期: 2024-06-12
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