响应面法优化泡叶藻专用复合酶体系构建工艺

Optimization of the construction process of a specialized composite enzyme system for Ascophyllum nodosum by response surface methodology

中文关键词:  泡叶藻  海藻酸钠  褐藻胶裂解酶  复合酶制剂  褐藻寡糖

英文关键词:Ascophyllum nodosum  sodium alginate  alginate lyase  composite enzyme preparation  alginate oligosaccharides

中文摘要:

      目的 研究褐藻胶裂解酶的最佳应用条件, 构建针对泡叶藻(Ascophyllum nodosum)的专用复合酶制剂最佳工艺。方法 通过控制变量法确定褐藻胶裂解酶的最适条件, 从褐藻胶裂解酶的最适条件出发, 以泡叶藻为研究对象, 以纤维素酶、果胶酶、木瓜蛋白酶作为复配酶, 通过泡叶藻酶解响应面实验, 获得泡叶藻复合酶制剂的最佳工艺条件, 以该条件对不同地理群体的泡叶藻(秘鲁、智利、法国)进行酶解, 以褐藻寡糖提取率为指标进行分析验证该复合酶制剂对不同来源的泡叶藻的降解作用, 比较酶解产物褐藻寡糖提取率及固体转化率, 验证该复合酶制剂的酶解效果。结果 褐藻胶裂解酶的最佳应用条件为加酶量0.50%, pH=7.5, 作用时间18 h, 酶解温度45 ℃, 泡叶藻专用复合酶制剂最佳工艺条件为纤维素酶1.82%, 果胶酶1.2%, 木瓜蛋白酶1.5%, 褐藻胶裂解酶0.6%, 料液比1:25 (g:mL), 酶解温度45 ℃, pH=7.22, 作用时间18 h, 在此条件下, 褐藻寡糖提取率为14.46%。结论 通过利用本研究复合酶制剂对不同来源的泡叶藻的降解作用分析, 发现该复合酶制剂对不同来源的泡叶藻均具有有效降解作用且该复合酶制剂酶解效果要高于现有市售商业复合酶制剂。本研究构建的复合酶可直接酶解泡叶藻制备褐藻寡糖, 实现了海藻资源的直接利用, 为低成本制备功能性褐藻寡糖及相关农用生物制品的开发提供一了种新思路与技术参考。

英文摘要:

      Objective To study the optimal application conditions for alginate lyase and construction the optimal process for a specialized composite enzyme preparation targeting Ascophyllum nodosum. Methods The optimal reaction conditions of alginate lyase were determined by the control variable method. Based on these optimal conditions, Ascophyllum nodosum was selected as the research material, and cellulase, pectinase and papain were used for composite enzyme preparation. The optimal technological parameters for compound enzymatic hydrolysis of Ascophyllum nodosum were obtained via response surface methodology. Under the optimized conditions, Ascophyllum nodosum from different geographical populations (Peru, Chile and France) were subjected to enzymatic hydrolysis. Using the extraction rate of alginate oligosaccharides as the evaluation index, the degradation capacity of the compound enzyme preparation against seaweed from different origins was analyzed. By comparing the extraction rate of alginate oligosaccharides and the solid conversion rate of enzymatic hydrolysis products, the hydrolysis efficiency of the composite enzyme preparation was verified. Results The optimal application conditions for the alginate lyase were an enzyme dosage of 0.50%, pH 7.5, reaction time 18 h and hydrolysis temperature 45 ℃. The optimal process conditions for the composite enzyme preparation of Ascophyllum nodosum enzymatic hydrolysis were cellulase 1.82%, pectinase 1.2%, papain 1.5% and alginate lyase 0.6%, solid-liquid ratio was 1:25 (g:mL), with an enzymatic hydrolysis temperature of 45 ℃, pH 7.22 and a reaction time of 18 h. Under these conditions, the extraction rates of alginate oligosaccharides was 14.46%. Conclusion By analyzing the degradation effects of the composite enzyme preparation from this study on Ascophyllum nodosum from different sources, it is found that this composite enzyme preparation effectively degrades Ascophyllum nodosum from various sources, and its enzymatic efficacy surpasses that of commercially available composite enzyme preparations on the market. The composite enzyme preparation constructed in this study can directly enzymolyze Ascophyllum nodosum to prepare alginate oligosaccharides, realizing the direct utilization of seaweed resources. It provides a new idea and technical reference for the low-cost preparation of functional alginate oligosaccharides and the development of related agricultural biological products.