羊骨抗氧化肽酶解法制备和响应面工艺优化

doi:10.3969/j.issn.1673-1689.2021.03.003

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羊骨抗氧化肽酶解法制备和响应面工艺优化
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                        10.3969/j.issn.1673-1689.2021.03.003
                    
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Optimization of Enzymatic Preparation of Antioxidant Peptides from Sheep Bone by Response Surface Methodology

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研究了羊骨抗氧化肽酶法提取工艺及其氨基酸组成。在单因素实验基础上，通过响应面分析法优化羊骨抗氧化肽的提取工艺，利用超滤分离获取不同截留相对分子质量的肽段，采用水杨酸法、DPPH法和铁离子还原法研究梯级肽的抗氧化活性并分析其氨基酸组成。结果表明，碱性蛋白酶为最优酶解蛋白酶，最佳酶解条件为pH 8.9、酶添加量6 400 U/g、酶解时间3.25 h、底物（骨粉）质量分数8.8%、酶解温度50 ℃。在此条件下，酶解液的·OH清除率、DPPH·清除率和Fe³⁺还原能力分别为（91.31±0.48）%、（32.00±0.12）%和0.085±0.003。通过超滤分级制备得到的组分Ⅰ（M_r≤3 000）、组分Ⅱ（3 000<M_r≤10 000）、组分Ⅲ（M_r>10 000）均具有抗氧化能力。组分Ⅰ抗氧化活性最强，必需氨基酸和疏水氨基酸质量分别占总氨基酸质量的30.41%和43.13%。羊骨抗氧化肽可作为潜在的天然抗氧化剂来源。

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Antioxidant peptides from sheep bone were prepared by enzymatic method, and the amino acids compositions of peptides were analyzed. On the basis of single factor test, the extraction conditions of antioxidant peptides from sheep bone were optimized using response surface methodology. At the same time, peptides with different retention molecular weights were isolated by ultrafiltration, and their antioxidant activity and amino acids compositions were detected. Salicylic acid method, 2,2-diphenyl-1-picrylhydrazyl (DPPH) method and deoxidization Fe³⁺ method were applied to investigate the antioxidant activity of peptides with gradient molecular weights and their amino acid compositions were also analyzed. The results showed that the optimum enzyme for enzymatic hydrolysis of sheep bone was alkaline protease, and the optimum process for polypeptide preparation was as follows: pH was 8.9, the amount of enzyme added was 6 400 U/g, hydrolysis duration was 3.25 h, substance mass fraction was 8.8% and the hydrolysis temperature was 50 ℃. The ·OH scavenging activity, DPPH· scavenging activity and the reducing power of the optimal hydrolysis products were (91.31±0.48)%, (32.00±0.12)% and (0.085±0.003), respectively. Components ⅠM_r≤3 000), Ⅱ(3 000<M_r≤10 000) and Ⅲ(M_r>10 000) obtained through ultrafiltration had antioxidant capacity. In addition, component Ⅰ(M_r≤3 000) had the highest antioxidant activity, in which the essential and hydrophobic amino acids accounted for 30.41% and 43.13% of the total amino acids, respectively. The antioxidant peptides from sheep bone can be explored as a potential natural antioxidant.

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张靖,苏琳,陈晓雨,要铎,赵丽华,王英丽,张忠海,靳烨.羊骨抗氧化肽酶解法制备和响应面工艺优化[J].食品与生物技术学报,2021,40(3):18-27.

ZHANG Jing, SU Lin, CHEN Xiaoyu, YAO Duo, ZHAO Lihua, WANG Yingli, ZHANG Zhonghai, JIN Ye. Optimization of Enzymatic Preparation of Antioxidant Peptides from Sheep Bone by Response Surface Methodology[J]. Journal of Food Science and Biotechnology,2021,40(3):18-27.

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在线发布日期: 2021-07-01
出版日期: 2021-03-25

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