扇贝蛋白质提取工艺的优化

Optimization of protein extraction process from Pectinidae

中文关键词:  扇贝  蛋白质  分段酶解  提取率

英文关键词:Pectinidae  protein  segmented enzymatic hydrolysis  extraction rate

中文摘要:

      目的 优化扇贝蛋白质提取工艺。方法 利用不同蛋白酶的酶切作用方式及酶切位点的不同, 采用复合酶与分段酶解相结合。利用单因素结合Plackett-Burman筛选出显著因素和Box-Behnken模型优化一段蛋白质提取工艺, 并通过正交实验优化二段酶解工艺参数。结果 复合酶分段酶解提取蛋白质的最佳条件为: 一段最佳提取工艺参数为: 固液比1:3.9 (m:V)、酶解时间80.0 min、复合酶加酶量1.1%、复合酶配比(胰蛋白酶:碱性蛋白酶)1:2 (m:m)、酶解pH 8.2、酶解温度52.5 ℃; 二段最佳提取工艺参数为: 酶解pH 7.0、酶解温度50.0 ℃、中性蛋白酶添加量0.8%、酶解时间40 min。在此条件下, 蛋白质提取率为76.3%。结论 由于复合酶同步和分段酶解形成协同效应和互补作用, 扇贝蛋白质提取率得到有效提高, 为后续扇贝蛋白质的开发和利用提供了一定的理论依据。

英文摘要:

      Objective To optimize the protein extraction process of Pectinidae. Methods A combination of composite enzymes and segmented enzymatic hydrolysis was employed, utilizing different protease enzymes with mode of enzyme digestion and cleavage sites. Significant factors were screened by single factor combined with Plackett-Burman and Box-Behnken model was used to optimize the one-stage protein extraction process, and the parameters of the two-stage enzymatic digestion process were optimized by orthogonal tests. Results The optimal extraction conditions for complex enzyme-assisted stepwise hydrolysis in protein extraction were as follows: The optimal extraction process parameters of the one-stage were as follows, solid-liquid ratio was 1:3.9 (m:V), enzyme digestion time was 80.0 min, compound enzyme addition was 1.1%, complex enzyme ratio (trypsin:alkaline protease) was 1:2 (m:m), enzyme digestion pH was 8.2, enzyme digestion temperature was 52.5 ℃; the optimal extraction parameters of the two-stage were as follows: Enzyme digestion pH was 7.0, enzyme digestion temperature was 50.0 ℃, neutral protease addition was 0.8%, and enzyme digestion time was 40 min. Under these conditions, the protein extraction rate was 76.3%. Conclusion Due to synergistic and complementary effects between the complex enzymes and segmented enzymatic hydrolysis, the protein extraction rate of Pectinidae is effectively improved, which provide some theoretical basis for the subsequent development and utilization of proteins from Pectinidae.