玉米蛋白环（组氨酸-脯氨酸）二肽前体的制备及其鉴定

doi:10.7506/spkx1002-6630-201622008

摘要/Abstract

摘要： 以玉米蛋白为原料，制备活性环（组氨酸-脯氨酸）二肽（cyclo（His-Pro））前体——His-Pro和Pro-His二肽。分别采用碱性蛋白酶、复合蛋白酶、风味蛋白酶和木瓜蛋白酶对提取的玉米γ-醇溶蛋白进行单酶和双酶分步水解，筛选出最佳酶解工艺。采用碱性蛋白酶和风味蛋白酶分步水解的水解产物中cyclo（His-Pro）前体的含量比其他水解产物要高。采用响应面和L16（45）正交试验对碱性蛋白酶和风味蛋白酶分步水解γ-醇溶蛋白的工艺进行了优化，最佳水解条件为：底物质量浓度32 mg/mL，先加入12 100 U/g的碱性蛋白酶水解6.5 h，再加入17 000 U/g的风味蛋白酶于pH 7.5、55 ℃条件下水解6 h。采用超高效液相色谱法和电喷雾质谱法对cyclo（His-Pro）前体进行定量和定性分析，结果表明水解产物中含有较高含量的脯氨酸-组氨酸二肽（6.88 mg/g）。

关键词: 玉米蛋白, 酶解, 环（组氨酸-脯氨酸）二肽前体, γ-醇溶蛋白

Abstract: The value-added use of corn protein for production of the cyclo (His-Pro) precursors histidyl-proline (His-Pro) and prolyl-histidine (Pro-His) was investigated. The extracted γ-zein was hydrolyzed with single proteases (alcalase, protamex, flavourzyme and papain) and two proteases in sequence, respectively. The hydrolysate prepared with alcalase and flavourzyme in sequence exhibited the highest content of cyclo (His-Pro) precursors. Response surface methodology and an orthogonal array design L16 (45) were applied to optimize the hydrolysis conditions. The optimum conditions for alcalase hydrolysis were as follows: substrate concentration of 32 mg/mL, enzyme/substrate (E/S) ratio of 12 100 U/g and hydrolysis time of 6.5 h, while the optimum conditions for flavourzyme hydrolysis were as follows: E/S ratio of 17 000 U/g, pH of 7.5, hydrolysis temperature of 55 ℃ and hydrolysis time of 6 h. Identification and quantification of cyclo (His-Pro) precursors in the hydrolysate were achieved by ultraperformance liquid chromatography (UPLC) and electrospray ionizationmass spectrometry (ESI-MS). The results showed that the hydrolysate contained a high content of Pro-His dipeptide (6.88 mg/g).

Key words: corn protein, enzymatic hydrolysis, cyclo (His-Pro) precursor, γ-zein

中图分类号:

Q516

张艳荣,樊红秀,刘鸿铖,董欣. 玉米蛋白环（组氨酸-脯氨酸）二肽前体的制备及其鉴定[J]. 食品科学, 2016, 37(22): 52-59.

ZHANG Yanrong, FAN Hongxiu, LIU Hongcheng, DONG Xin. Preparation and Identification of Cyclo (His-Pro) Precursors from Corn Protein[J]. FOOD SCIENCE, 2016, 37(22): 52-59.

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