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用SDS-PAGE比较三种商业酶对两种底物(米糠浓缩蛋白和大豆蛋白)进行酶解的效果。

Comparison the effect of three commercial enzymes for enzymatic hydrolysis of two substrates (rice bran protein concentrate and soy-been protein) with SDS-PAGE.

作者信息

Ahmadifard Nasrollah, Murueta Julio Humberto Cordova, Abedian-Kenari Abdolmohammad, Motamedzadegan Ali, Jamali Hadi

机构信息

Fisheries Department, Faculty of Natural Resources, Urmia University, P.O. Box: 57153-165, Urmia, Iran.

Centro de Investigaciones Biológicas del Noroeste (CIBNOR), B.C.S. 23090 La Paz, Mexico.

出版信息

J Food Sci Technol. 2016 Feb;53(2):1279-84. doi: 10.1007/s13197-015-2087-6. Epub 2015 Nov 9.

Abstract

In this research enzymatic hydrolysis of rice bran protein concentrate (RBPC) and soybean Protein (SBP) as control were studied with 3 commercial enzymes (Alcalase®, Papain and acommercial 3-enzyme cocktail containing of 1.6 mg ml(-1) Trypsin, 3.1 mg ml(-1) Chymotrypsin, 1.3 mg ml(-1)Aminopeptidase (SIGMA P7500) and 7.95 mg ml(-1)pronase type XIV (SIGMA P5147) by the pH stat method. The hydrolysis was carried out at temperature of 28 C, 60 min and pH 8.00. Results were showed that RBPC, and SBP had higher Degree hydrolysis (DH %) with Alcalase® enzyme. Alcalase®had stronger capability for hydrolysis compared to the other tested enzymes. After 60 minute of hydrolysis time, the DH% of Alcalase® for RBPC and SBP was 12.69 and 12.50 %, respectively. In contrast, papain enzyme was showed lowest DH% in three substrates that 1.56 and 1.24 % were for SBP and RBPC, respectively.The hydrolysis of the protein fraction performed the three enzymes on the two substrates was followed in SDS-PAGE. RBPC and SBP showed almost complete digestion with Alcalase® enzyme after 60 minutes. 3-enzyme cocktail enzyme hydrolyzed better the RBPC than the SBP. Papain enzyme had less effect on the two substrates than other 2 enzymes. It was found that Alcalase® has highest capability for hydrolysis compared to other enzyme preparations. The high value protein hydrolysates prepared by Alcalase® can be used as value added ingredients in many food formulations. They are also suitable for a broad range of industrial food applications and also for cosmetic and personal care products.

摘要

在本研究中,采用pH计法,用3种商业酶(碱性蛋白酶、木瓜蛋白酶和一种含有1.6mg/ml胰蛋白酶、3.1mg/ml胰凝乳蛋白酶、1.3mg/ml氨肽酶(SIGMA P7500)和7.95mg/ml十四型链霉蛋白酶(SIGMA P5147)的商用3酶混合物)对米糠浓缩蛋白(RBPC)和作为对照的大豆蛋白(SBP)进行酶解。酶解在28℃、60分钟和pH8.00的条件下进行。结果表明,RBPC和SBP在碱性蛋白酶作用下具有较高的水解度(DH%)。与其他测试酶相比,碱性蛋白酶具有更强的水解能力。水解60分钟后,碱性蛋白酶对RBPC和SBP的DH%分别为12.69%和12.50%。相比之下,木瓜蛋白酶在三种底物中显示出最低的DH%,SBP和RBPC的DH%分别为1.56%和1.24%。通过SDS-PAGE跟踪了三种酶对两种底物的蛋白质部分的水解情况。60分钟后,RBPC和SBP在碱性蛋白酶作用下几乎完全消化。3酶混合物对RBPC的水解效果优于SBP。木瓜蛋白酶对两种底物的作用小于其他两种酶。结果发现,与其他酶制剂相比,碱性蛋白酶具有最高的水解能力。由碱性蛋白酶制备的高价值蛋白质水解产物可作为增值成分用于许多食品配方中。它们也适用于广泛的工业食品应用以及化妆品和个人护理产品。

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