羊栖菜寡糖的酶法制备及其抗氧化性与抑菌活性研究

Study on preparation of Sargassum fusiforme oligosaccharides by enzyme and its antioxidant and antibacterial properties

  • 摘要: 羊栖菜多糖 (Sargassum fusiforme polysaccharides) 是羊栖菜中含量最丰富且极具特色的有效成分。海洋寡糖是由海洋多糖降解而来的产物。为从羊栖菜中提取活性较高的寡糖,并探究其抗氧化性及抑菌活性,以水提法所得的羊栖菜多糖为原料,利用褐藻胶裂解酶对其进行酶解,制备羊栖菜寡糖。通过单因素和正交试验,对酶解工艺进行优化,考察底物质量分数、加酶量和酶解时间3个因素对羊栖菜寡糖还原糖含量及平均聚合度的影响,确定酶解羊栖菜寡糖的最佳工艺参数为:酶解时间48 h、底物质量分数1.5%、加酶量20 U·mL−1,测得还原糖的质量浓度为 (26.53±0.92) mg·mL−1。经薄层层析法分析所制备寡糖的主要成分为单糖、双糖和三糖。抗氧化性和抑菌活性的测定结果表明:当质量浓度为3.0 mg·mL−1时,寡糖对DPPH (1,1-二苯基-2-三硝基苯肼)、ABTS 2,2'-联氮-双 (3-乙基苯噻唑啉-6-磺酸) 二铵盐、羟自由基和超氧阴离子自由基的清除率分别达到63.1%、80.7%、71.3%和57.2%,当质量浓度为3.5 mg·mL−1时,寡糖的还原力为0.923。抑菌活性结果表明,当羊栖菜寡糖质量浓度在10和100 mg·mL−1时,对白色念珠菌 (Candida albicans)、金黄色葡萄球菌 (Staphylococcus aureus)、大肠埃希菌 (Escherichia coli)、铜绿假单胞菌 (Pseudomonas aeruginosa)均具有较好的抑菌作用。相对于羊栖菜多糖,羊栖菜寡糖在抗氧化性和抑菌活性上均有更好的效果。

     

    Abstract: Sargassum fusiforme polysaccharides are the most abundant and extremely distinctive ingredients in S. fusiforme. Marine oligosaccharides are products derived from the degradation of marine polysaccharides. To extract more active oligosaccharides from S. fusiforme, we investigated the effects of oligosaccharides on antioxidant and antibacterial properties. We took S. fusiforme polysaccharide through water extraction as the raw material and prepared it by using enzymatic hydrolysis with fucoidan lyase. Then, we conducted single factor and orthogonal trials to optimize the enzymatic hydrolysis process. We examined the average degree of polymerization and reduced sugar content of S. fusiforme oligosaccharides in relation to substrate concentration, enzyme dosage, and enzymatic hydrolysis time. The optimal process parameters for enzymatic hydrolysis of S. fusiforme oligosaccharides were as follows: enzymatic hydrolysis time of 48 h, substrate concentration of 1.5%, enzyme dosage of 20 U·mL−1, and a measured reducing sugar content of (26.53±0.92) mg·mL−1. Thin layer chromatography was used to produce the monosaccharides, disaccharides and trisaccharides. The antioxidant and antibacterial properties of oligosaccharides were determined: when the mass volume concentration was 3.0 mg·mL−1, the scavenging rates of DPPH, ABTS, hydroxyl radical and superoxide anion radical were 63.1%, 80.7%, 71.3% and 57.2%, respectively. When the mass volume concentration was 3.5 mg·mL−1, the reducing power of oligosaccharides was 0.923. The bacteriostatic results show that S. fusiforme oligosaccharides have good bacteriostatic impact on Candida albicans, Staphylococcus aureus, Escherichia coli and Pseudomonas aeruginosa. Compared with S. fusiforme polysaccharide, it shows enhanced antioxidant and antibacterial effects.

     

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