食品科学 ›› 2017, Vol. 38 ›› Issue (14): 193-199.doi: 10.7506/spkx1002-6630-201714030

• 工艺技术 • 上一篇    下一篇

响应面法优化中性蛋白酶提取苦竹花多糖及多糖性质分析

葛雪筠,周德健,王斌,陈荫   

  1. (浙江海洋大学食品与医药学院,浙江?舟山 316022)
  • 出版日期:2017-07-25 发布日期:2017-09-06
  • 基金资助:
    国家自然科学基金青年科学基金项目(41406142);浙江省自然科学基金项目(21136000114)

Optimization of Neutroproteinase-Assistant Extraction of Polysaccharide from the Stroma of Shiraia bambusicola Henn by Response Surface Analysis and Characterization of the Extracted Polysaccharide

GE Xuejun, ZHOU Dejian, WANG Bin, CHEN Yin   

  1. (College of Food and Pharmacy, Zhejiang Ocean University, Zhoushan 316022, China)
  • Online:2017-07-25 Published:2017-09-06

摘要: 目的:采用响应面法优化中性蛋白酶辅助提取苦竹花多糖的最优条件,并对提取得到的粗多糖进行分离纯化、理化性质及体外抗氧化性测定。方法:在单因素试验基础上,考察提取时间、酶添加量、提取温度对苦竹花粗多糖得率的影响。利用响应面试验建立数学模型,确定最佳提取条件。苦竹花粗多糖经分离纯化,获取2?个组分,分别命名为SBH-1和SBH-2,其中以SBH-1为主要组分。通过高效凝胶渗透色谱和红外光谱对SBH-1组分进行单糖组成、分子质量及初步的结构研究。通过与VC对比,研究苦竹花粗多糖SBH-1的体外抗氧化活性,即1,1-二苯基-2-三硝基苯肼(1,1-diphenyl-2-picrylhydrazyl,DPPH)自由基清除率、超氧阴离子自由基清除率及抗脂质过氧化能力的测定。结果:在中性蛋白酶添加量0.83%,提取温度46.68?℃,提取时间115.95?min条件下,粗多糖得率为7.63%。经检测苦竹花粗多糖SBH-1组分分子质量为18.3?kD,主要由甘露糖(Man)、葡萄糖(Glc)、半乳糖(Gal)3?种单糖组成比例为1.3∶2.3∶1.2。SBH-1对DPPH自由基和超氧阴离子自由基清除EC50分别为2.03?mg/mL和1.038?mg/mL。结论:采用中性蛋白酶辅助提取得到的苦竹花粗多糖简单易行,纯化得到的中性杂多糖SBH-1具有很好的活性,为进一步研究和开发提供理论基础。

关键词: 苦竹花, 多糖, 响应面, 体外抗氧化

Abstract: Objective: This study aimed to optimize the neutroproteinase-assisted extraction of polysaccharides from the stroma of Shiraia bambusicola Henn using response surface methodology (RSM) and to investigate the separation, purification, characterization and antioxidant activity of the polysaccharides. Methods: The effects of extraction time, enzyme concentration and extraction temperature on the yield of polysaccharides were examined by using one-factor-at-a-time experiments. These extraction conditions were optimized through mathematical modeling. The crude extract was fractionated into two fractions: SBH-1 and SBH-2, with SBH-1 being among them. The monosaccharide composition, molecular mass and structure of SBH-1 were detected by high performance gel permeation chromatography (HPGPC) and infrared spectroscopy, respectively. The in vitro antioxidant activity was measured in comparison with VC by superoxide anion scavenging, 1,1-diphenyl-2-picrylhydrazyl (DPPH) scavenging, and lipid peroxidation inhibition assays. Results: The maximum yield of crude polysaccharides of 7.63% was obtained at 115.95 min of extraction at 46.68 ℃ with 0.83% neutral proteinase. The molecular mass of SBH-1 was determined to be 18.3 kD, and its monosaccharide composition consisted of glucose, mannose, and galactose with a ratio of 1.3:2.3:1.2. The EC50 values for scavenging of DPPH and superoxide anion radials were 2.03 and 1.038 mg/mL respectively. Conclusions: The neutroproteinase-assistant extraction procedure was simple and easy to operate. SBH-1 was a neutral heteropolysaccharide with high activity. This study can provides a theoretical basis for further research and development of polysaccharides from S. bambusicola Henn.

Key words: Shiraia bambusicola Henn, polysaccharide, response surface methodology, antioxidant activity

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