鲣鱼黄嘌呤氧化酶抑制肽酶法制备工艺优化

doi:10.3785/j.issn.1008-9209.2019.01.101

浙江大学学报（农业与生命科学版）

2019, Vol. 45

Issue (5): 550-562 DOI: 10.3785/j.issn.1008-9209.2019.01.101

食品科学

鲣鱼黄嘌呤氧化酶抑制肽酶法制备工艺优化

邹琳¹(

),杭妙佳¹,李阳¹,杜鹃²,冯凤琴¹(

)

1. 浙江大学生物系统工程与食品科学学院，杭州 310058
2. 杭州康源食品科技有限公司，杭州 310003

Optimization of enzymatic hydrolysis process for preparing xanthine oxidase inhibitory peptides from skipjack tuna

Lin ZOU¹(

),Miaojia HANG¹,Yang LI¹,Juan DU²,Fengqin FENG¹(

)

1. College of Biosystems Engineering and Food Science, Zhejiang University, Hangzhou 310058, China
2. Hangzhou Kangyuan Food Science & Technology Co. , Ltd. , Hangzhou 310003, China

全文: PDF(4822 KB) HTML

摘要：

以鲣鱼为原料，利用酶法制备鲣鱼黄嘌呤氧化酶（xanthine oxidase, XOD）抑制肽。在单因素试验的基础上，采用Box-Behnken响应面分析法，以水解度、氮回收率、XOD抑制活性、肌肽及鹅肌肽含量为响应值，对酶解工艺中蛋白酶种类、加酶量、酶解pH、温度和时间等影响因素进行优化，得到酶解的最优工艺：采用中性蛋白酶，加酶量为489.86 U/g，酶解pH为7.08，酶解温度为49.5 ℃，酶解时间为5 h。在此条件下，鲣鱼酶解产物的水解度为22.38%，氮回收率为83.31%，XOD抑制活性达到62.26%，肌肽和鹅肌肽含量分别为0.05%和2.45%（以干基计），与回归模型理论预测值基本一致。由本法所得的鲣鱼XOD抑制肽是以分子质量低于1 000 Da的寡肽为主。皮尔逊相关性分析表明，XOD抑制活性与肌肽或鹅肌肽含量无相关性，推测在酶解过程中产生了其他具有XOD抑制活性的小分子肽。

关键词： 鲣鱼; 黄嘌呤氧化酶抑制肽; 酶解工艺; 肌肽; 鹅肌肽

Abstract:

Skipjack tuna was enzymatically hydrolyzed to prepare xanthine oxidase (XOD) inhibitory peptides. The enzymatic hydrolysis process was optimized by investigating the effects of protease types and enzymatic dosages, pH, temperature, time and their interactions on degree of hydrolysis, nitrogen recovery, XOD inhibition activity and the contents of carnosine and anserine using one-factor-at-a-time method and response surface methodology. The optimal process parameters were as follows: the application of neutrase with the dosage of 489.86 U/g, the pH of 7.08, the temperature of 49.5 ℃, and the time of 5 h. Under the optimal conditions, the degree of hydrolysis was 22.38%; nitrogen recovery was 83.81%; XOD inhibition activity was 62.26%; and the contents of carnosine and anserine were 0.05% and 2.45% (on a dried basis), respectively, which were all in good agreement with the predicted values. The XOD inhibitory peptides obtained were mainly composed of a fraction with molecular mass of less than 1 000 Da. The Pearson’s correlation analysis showed that the XOD inhibition activity of dorsal and ventral muscle in skipjack tuna was not associated with the contents of carnosine and anserine, and it may be due to the production of other XOD inhibitory peptides during the enzymatic hydrolysis.

Key words: skipjack tuna xanthine oxidase inhibitory peptides enzymatic hydrolysis process carnosine anserine

收稿日期: 2019-01-10 出版日期: 2019-12-05

CLC:

TS 254.1

基金资助: 浙江大学科技合作项目“水产动物源功能蛋白肽产品研究开发”(K横20182914);浙江大学实验技术研究项目(SJS201708)

通讯作者: 冯凤琴 E-mail: zoulin@zju.edu.cn;fengfq@zju.edu.cn

作者简介: 邹琳（https://orcid.org/0000-0002-4892-251X），E-mail：zoulin@zju.edu.cn

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引用本文:

邹琳,杭妙佳,李阳,杜鹃,冯凤琴. 鲣鱼黄嘌呤氧化酶抑制肽酶法制备工艺优化[J]. 浙江大学学报（农业与生命科学版）, 2019, 45(5): 550-562.

Lin ZOU,Miaojia HANG,Yang LI,Juan DU,Fengqin FENG. Optimization of enzymatic hydrolysis process for preparing xanthine oxidase inhibitory peptides from skipjack tuna. Journal of Zhejiang University (Agriculture and Life Sciences), 2019, 45(5): 550-562.

链接本文:

http://www.zjujournals.com/agr/CN/10.3785/j.issn.1008-9209.2019.01.101 或 http://www.zjujournals.com/agr/CN/Y2019/V45/I5/550

表1 各蛋白酶的最适酶解条件

表2 不同蛋白酶对XOD抑制活性、氮回收率和水解度的影响

图1 加酶量对鲣鱼背腹肉酶解效果的影响短栅上的不同小写字母表示在P＜0.05水平差异有统计学意义；n=3。

图2 酶解温度对鲣鱼背腹肉酶解效果的影响短栅上的不同小写字母表示在P＜0.05水平差异有统计学意义；n=3。

图3 酶解pH对鲣鱼背腹肉酶解效果的影响短栅上的不同小写字母表示在P＜0.05水平差异有统计学意义；n=3。

图4 酶解时间对鲣鱼背腹肉酶解效果的影响短栅上的不同小写字母表示在P＜0.05水平差异有统计学意义；n=3。

表3 酶解工艺优化的响应面试验因素与水平

表4 酶解工艺优化的响应面试验设计及结果

表5 肌肽、鹅肌肽含量与XOD抑制活性的皮尔逊相关性分析

图6 鲣鱼背腹肉酶解产物分子质量分布情况

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