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Journal of Zhejiang University SCIENCE B
ISSN 1673-1581(Print), 1862-1783(Online), Monthly
2017 Vol.18 No.8 P.674-684
Ultrasound extraction optimization, structural features, and antioxidant activity of polysaccharides from Tricholoma matsutake
Abstract: An ultrasonic-assisted technique was employed to extract crude polysaccharide from Tricholoma matsutake fruiting bodies. Single-factor tests and orthogonal experimental design (L9(33)) were used to obtain the optimal extraction conditions. Results showed that the optimal parameters were as follows: ultrasonic temperature, 40 °C; ultrasonic time, 50 min; water to raw material ratio, 25 ml/g; ultrasonic frequency, 45 kHz; and ultrasonic power, 100 W. Three novel T. matsutake polysaccharide (TMP) fractions (TMP30, TMP60, and TMP80) were isolated and purified from TMP by stepwise alcohol precipitation. Their preliminary structural features were determined by high-performance anion-exchange chromatography with pulsed-amperometric detection (HPAEC-PAD) and Fourier transform infrared spectrophotometer (FT-IR) analyses. Furthermore, their in vitro antioxidant activity was investigated in terms of a reducing power assay and the scavenging rates of 2,2-diphenyl-1-picrylhydrazyl (DPPH) and hydroxyl radicals. The order of the various fractions based on their antioxidant activity was TMP80>TMP>TMP60>TMP30. These findings suggested that novel polysaccharide fractions from T. matsutake, especially TMP80, could be promising active macromolecules for biomedical use.
Key words: Tricholoma matsutake polysaccharide; Orthogonal test design; Ultrasound-assisted extraction; Monosaccharide composition; Antioxidant activity
创新点：首次对超声辅助制备的TMP进行了乙醇分级，对其进行了结构和抗氧化活性评估，并筛选出了抗氧化活性最强的部位（TMP80）。
方法：采用单因素试验和正交试验（L9(33)）对超声提取TMP的工艺条件进行了优化；采用不同的乙醇终浓度对TMP分离纯化为3种多糖TMP30、TMP60和TMP80；采用高效阴离子色谱（HPAEC-PAD）和傅里叶红外（FT-IR）等技术对3种多糖的理化性质进行了结构表征；利用还原力测定、二苯三硝基苯肼（DPPH）和羟自由基的清除作用对它们的抗氧化活性进行了评估。
结论：正交试验结果表明，超声提取TMP的最优工艺条件为：超声温度40 °C，超声时间50 min，水原料比例25 ml/g，超声波频率45 kHz和超声波功率100 W（表1和表2）。在此条件下的TMP得率为8.06%。采用乙醇分级法对TMP进一步分离纯化，制备得到3种多糖TMP30、TMP60和TMP80。HPAEC-PAD和FT-IR的结果表明，TMP30、TMP60和TMP80均主要由岩藻糖（L-Fuc）、半乳糖（D-Gal）、葡萄糖（D-Glc）、木糖（D-Xyl）和甘露糖（D-Man）组成，它们具有相同的单糖组成，但含有不同的摩尔比，且均有β-构型（图2、图3和表3）。抗氧化活性结果表明，它们的能力高低顺序为：TMP80>TMP>TMP60>TMP30（图4和表4），TMP80抗氧化活性最高，为松茸多糖的进一步开发利用提供重要的科学依据。
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DOI:
10.1631/jzus.B1600239
CLC number:
R284.2
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On-line Access:
2017-08-08
Received:
2016-05-24
Revision Accepted:
2016-09-12
Crosschecked:
2017-07-19