Study on enzymatic hydrolysis of soybean β-conglycinin using alkaline protease from Bacillus subtilis ACCC 01746 and antigenicity of its hydrolysates

Q2 Agricultural and Biological Sciences Grain Oil Science and Technology Pub Date : 2021-03-01 DOI:10.1016/j.gaost.2020.12.001
Haicheng Yin, Xinrui Zhang, Jin Huang
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引用次数: 8

Abstract

Due to its beneficial health effects, the use of soybean protein has shown a continuous increase, but concerns regarding the allergenicity of soybean antigenic protein have also increased. This study aimed to evaluate the hydrolytic effects of a non-commercial alkaline protease isolated from the Bacillus subtilis ACCC 01746 on soybean β-conglycinin and the allergenicity of its hydrolysates. Alkaline protease of the strain was separated by precipitation method of organic solvents, and the β-conglycinin was separated by alkali-solution and acid-isolation and purified by use of gel column. Using the degree of hydrolysis (DH) and inhibition rate as evaluation indexes, the enzymatic hydrolysis parameters of β-conglycinin was optimized by single factor and L9(34) orthogonal tests, so as to explore the effect of the protease on the hydrolysis degree and the antigenicity of β-conglycinin hydrolysates. The results showed that the native enzyme existed as an 18.3 kDa monomer with a 430 U/g maximum activity. The purity of β-conglycinin was 84.8%. The single-factor test results showed that DH showed the oppostie trendency with the inhibition rate, and the increase of protein concentration caused monotone increasing and monotone decreasing of the inhibition rate and the DH, and the optimal protein concentration was 30 mg/mL. The optimization results showed that pH had the largest impacts on both DH and the inhibition rate, followed by enzyme dosage, hydrolysis temperature and hydrolysis time. Under the optimum hydrolysis conditions of protein concentration 30 mg/mL, enzyme dosage 0.7%, hydrolysis time 40 min, temperature 55 °C and pH 8.5, the DH reached the highest of 76.28%, and the inhibition rate was the lowest of 27.03%, which was reduced greatly compared with that before optimization. These results suggested that alkaline protease appeared to show a relatively high effeciency in lowering soybean allergenicity, making it possible to produce low-allergenicity soybean protein.

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枯草芽孢杆菌ACCC 01746碱性蛋白酶水解大豆β-甘氨酸及其水解产物抗原性的研究
由于其有益健康的作用,大豆蛋白的使用不断增加,但对大豆抗原蛋白致敏性的关注也在增加。本研究旨在评价从枯草芽孢杆菌ACCC 01746中分离的一种非商业碱性蛋白酶对大豆β-甘氨酸的水解作用及其水解产物的致敏性。采用有机溶剂沉淀法分离菌株的碱性蛋白酶,采用碱液和酸分离法分离菌株的β-甘氨酸,并用凝胶柱纯化菌株。以水解度(DH)和抑制率为评价指标,通过单因素试验和L9(34)正交试验对β-甘氨酸的酶解参数进行优化,探讨蛋白酶对β-甘氨酸水解物水解度和抗原性的影响。结果表明,该天然酶以18.3 kDa单体存在,最大活性为430 U/g。β-甘氨酸纯度为84.8%。单因素试验结果表明,DH与抑制率呈相反趋势,蛋白浓度的增加导致抑制率和DH呈单调上升和单调下降趋势,最佳蛋白浓度为30 mg/mL。优化结果表明,pH对DH和抑制率的影响最大,其次是酶用量、水解温度和水解时间。在蛋白浓度为30 mg/mL、酶用量为0.7%、水解时间为40 min、温度为55℃、pH为8.5的最佳水解条件下,DH最高为76.28%,抑制率最低为27.03%,较优化前显著降低。由此可见,碱性蛋白酶具有较高的降低大豆致敏性的作用,为制备低致敏性大豆蛋白提供了可能。
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来源期刊
CiteScore
7.30
自引率
0.00%
发文量
69
审稿时长
12 weeks
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