中国农业科学 ›› 2014, Vol. 47 ›› Issue (15): 3044-3057.doi: 10.3864/j.issn.0578-1752.2014.15.014

• 贮藏·保鲜·加工 • 上一篇    下一篇

甘薯淀粉加工废渣制备复合寡糖的条件优化及其活性评价

 董向艳1, 李静梅2, 石波1, 彭晴1, 乔宇1, OjokohEromosele1, 张迷敏1   

  1. 1、中国农业科学院饲料研究所,北京 100081;
    2、中国农业科学院作物科学研究所,北京 100081
  • 收稿日期:2014-03-05 出版日期:2014-08-01 发布日期:2014-06-04
  • 通讯作者: 石波,Tel:010-82106075;E-mail:shibo@caas.cn
  • 作者简介:董向艳,E-mail:13810057864@163.com
  • 基金资助:

    “十二五”农村领域国家科技计划(2011BAD26B02-3)、现代农业产业技术体系建设专项基金(CARS-11-B-18)

Optimization and Functional Assessment of Oligosaccharides Compound Prepared by Sweet Potato Residue

 DONG  Xiang-Yan-1, LI  Jing-Mei-2, SHI  Bo-1, PENG  Qing-1, QIAO  Yu-1, OjokohEromosele1 , ZHANG  Mi-Min-1   

  1. 1、Feed Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081;
    2、Institute of Crop Sciences, Chinese Academy of Agricultural Sciences, Beijing 100081
  • Received:2014-03-05 Online:2014-08-01 Published:2014-06-04

摘要: 【目的】探索商品化β-葡聚糖酶和多聚半乳糖醛酸酶共同水解甘薯淀粉加工废渣(简称甘薯渣)制备复合寡糖的最佳条件,并利用复合寡糖诱导大豆生成大豆抗毒素,为复合寡糖的工业化生产及应用提供科学依据。【方法】分别用商品化β-葡聚糖酶、多聚半乳糖醛酸酶水解甘薯渣,以温度、pH、底物浓度、酶添加量和反应时间为条件开展单因素试验,利用TLC和HPLC对酶解产物进行测定,分别以纤维二糖得率、果胶二糖和果胶三糖总得率为指标得到单因素试验的最佳条件,再通过复合酶共同水解甘薯渣制备复合寡糖,并对纤维寡糖、果胶寡糖以及复合寡糖这3种寡糖产物进行诱导大豆抗毒素活性评价。【结果】纤维寡糖制备的单因素试验结果表明,当温度40℃、pH3.5、底物浓度1%、β-葡聚糖酶添加量6.9×103 U•g-1甘薯渣膳食纤维、反应时间7 h时酶解效果最好,寡糖产物以纤维二糖为主,纤维二糖得率为100.6 mg•g-1(纤维二糖质量/甘薯渣膳食纤维质量),纤维二糖转化率为22.37%(纤维二糖质量/甘薯渣膳食纤维中纤维素质量)。果胶寡糖制备的单因素试验结果表明,当温度40℃、pH2.5、底物浓度1%、多聚半乳糖醛酸酶添加量1.42×104 U•g-1甘薯渣膳食纤维、反应时间4 h时酶解效果最好,寡糖产物以果胶二糖和果胶三糖为主,果胶二糖和果胶三糖总得率为17.43 mg•g-1(果胶二糖与果胶三糖的总质量/甘薯渣膳食纤维质量),果胶二糖和果胶三糖总转化率为29.9%(果胶二糖与果胶三糖的总质量/甘薯渣膳食纤维中果胶质量)。根据上述单因素试验结果优化复合寡糖制备条件,在温度40℃、pH2.5、底物浓度1%、β-葡聚糖酶添加量6.9×103 U•g-1甘薯渣膳食纤维、多聚半乳糖醛酸酶添加量1.42×104 U•g-1甘薯渣膳食纤维、反应7 h时,复合寡糖产物中以纤维二糖、果胶二糖和果胶三糖为主,纤维二糖得率为136.97 mg•g-1,纤维二糖转化率为33.57%;果胶二糖和果胶三糖的总得率为25.96 mg•g-1,果胶二糖和果胶三糖总转化率为44.53%,与单一寡糖制备结果相比均有明显提高。利用甘薯复合寡糖作为外源诱导剂诱导大豆生成大豆抗毒素,当复合寡糖浓度为1%,大豆在无菌水中浸泡5 h,诱导温度25℃、湿度50%、黑暗中培养4 d时,大豆抗毒素生成量达到最高,为1.21 mg•g-1干豆重。而在相同条件下纤维寡糖和果胶寡糖诱导得到的大豆抗毒素生成量分别为0.80和0.46 mg•g-1干豆重。结果表明,甘薯复合寡糖对大豆抗毒素的诱导效果优于单一寡糖。【结论】甘薯渣成本低廉,可作为制备复合寡糖的优良原料,试验得到制备复合寡糖的最佳工艺条件,以其制备的复合寡糖对大豆抗毒素的生成与积累具有极佳的效果。

关键词: 甘薯渣, 纤维寡糖, 果胶寡糖, 复合寡糖, 酶解, 大豆抗毒素

Abstract: 【Objective】The aim of this study was to explore the optimum conditions of sweet potato residue degraded into oligosaccharides by commercialized β-glucanase and polygalacturonse and to investigate glyceollins accumulation in soybean elicited by oligosaccharides, thus providing a scientific basis for the industrial production and application of oligosaccharides.【Method】 Sweet potato residue was hydrolyzed by commercialized β-glucanase and polygalacturonse separately in single factor experiments for optimal conditions such as temperature, pH, substrate concentration, additive amount of enzyme and the reaction time of hydrolysis. Product obtained was determined by thin-layer-chromatography (TLC) and high performance liquid chromatography (HPLC). The optimal conditions for hydrolysis of sweet potato residue were obtained through analyzing the yield of cellobiose and the yield of digalacturonic acid and trigalacturonic acid. Then using the combi-enzyme hydrolyze the sweet potato residue to prepare oligosaccharides compound which could elicit glyceollins accumulation in soybean.【Result】The enzyme reaction conditions for the solubilization were optimized to be addition of 6.9×103 U β-glucanase by weight of sweet potato residue dietary fiber, 7 h treatment at pH3.5, temperature at 40℃, and substrate concentration at 1%. Under these conditions, cello-oligosaccharides were mainly cellobiose, the yield of cellobiose was 100.6 mg•g-1 sweet potato residue dietary fiber, and the conversion rate of cellobiose was 22.37%. The enzyme reaction conditions for the solubilization were optimized to be addition of 1.42×104 U polygalacturonse by weight of sweet potato residue dietary fiber, 4 h treatment at pH2.5, and temperature at 40℃. Using the optimum conditions, the yield of pectic oligosaccharides mainly with a degree of polymerization (DP) of 2 and 3, the yield of digalacturonic acid and trigalacturonic acid was 17.43 mg•g-1, the conversion rate of digalacturonic acid and trigalacturonic acid was 29.9%. The optimum conditions for producing oligosaccharides compound were as follows: temperature at 40℃, pH2.5, concentration of sweet potato residue at 1%, the additive amount of β-glucanase was 6.9×103 U•g-1 and the additive amount of polygalacturonse was 1.42×104 U•g-1, and the reaction time was 7 h. Using the optimum conditions, the yield of oligosaccharides compound mainly with a DP of 2 and 3, and the yield of cellobiose was 136.97 mg•g-1, the conversion rate of cellobiose was 33.57%, the yield of digalacturonic acid and trigalacturonic acid was 25.95 mg•g-1, the conversion rate of digalacturonic acid and trigalacturonic acid was 44.53%. Oligosaccharides compound was used to induce glyceollins synthesis in soybean seeds. The optimal conditions of elicitation were as follows: concentration of oligosaccharides compound 1%, presoaked in the sterile water for 5 h, stored at a controlled temperature of 25℃ and humidity of 50% in dark for 4 d, the yield of glyceollins elicited by oligosaccharides compound was 1.21 mg•g-1 dry weight soybeans. The yield of glyceollins elicited by cello-oligosaccharides with the same concentration of cellobiose was 0.80 mg•g-1 dry weight soybean. The yield of glyceollins elicited by pectic oligosaccharides under the same condition was 0.46 mg•g-1 dry weight soybeans.【Conclusion】 Sweet potato residue, as a cheap reaction substrate, could be used as the substrate for the production of oligosaccharides compound. The optimum conditions for the production of oligosaccharides compound were obtained, and the result showed that oligosaccharides compound has good effect on the generation and accumulation of glyceollins.

Key words: sweet potato residue , cello-oligosaccharides , pectic oligosaccharides , oligosaccharides compound , enzyme hydrolysis , glyceollins