响应面优化纤维素酶法提取茶花多糖工艺及其抗氧化活性研究
Study on the optimization of enzymatic extraction technology of polysaccharide from Camellia japonica L.by response surface methodology and investigation on its antioxidant activity
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摘要: 目的:优化纤维素酶提取茶花多糖的工艺,并评价其抗氧化活性。方法:以茶花多糖得率为响应值,在单因素实验基础上,以液料比、酶解温度、酶解时间、酶添加量为实验因素,采用响应面法建立数学模型,筛选最佳提取工艺条件;采用自由基清除能力体系评价茶花多糖的抗氧化活性。结果:通过二次回归模型响应面分析,影响茶花多糖得率的因素按主次顺序排列为:酶解时间>酶解温度>液料比>酶添加量;确定纤维素酶酶解茶花多糖最佳工艺条件为纤维素酶添加量5.0 mg/m L、液料比9∶1 m L/g、酶解温度48℃、酶解时间71 min,在此条件下茶花多糖得率为15.28%,模型方程理论预测值为15.91%,两者相对误差小于5%。茶花多糖具有较强的抗氧化活性,对DPPH和O-2·自由基的半数抑制浓度分别为0.974、1.342 mg/m L,但与维生素C比较,抗氧化活性较弱。结论:采用响应面法优化得到了茶花多糖的最佳提取工艺,该工艺方便可行,得到的多糖具有较强的抗氧化活性。Abstract: Objective: This study aimed to optimize the enzymatic extraction technology of polysaccharide from Camellia japonica L. and investigate its antioxidant activity. Methods: The response surface method( RSM) was adopted to establish the mathematical model. The polysaccharides extraction rate was used as response value,and the ratio of water to material,cellulase concentration,extraction temperature and extraction time were used as experimental factors to optimize the polysaccharide extraction conditions from Camellia japonica L.. Antioxidant activity of polysaccharides was measured by DPPH and O-2·free radical elimination method.Results: The optimum conditions obtained by RSM were as follows,polysaccharide yield was most significantly affected by extraction time,followed by extraction temperature,water to material ratio and cellulase concentration. The concentration of cellulase was 5.0 mg / m L,extraction time was 71 min,the ratio of water to feedstock was 9 ∶ 1 m L / g,extraction temperature was 48 ℃. Under the optimal conditions,the predicted extraction rate by mathematical model was15.91%,while the experimental extraction rate was 15.28%,with a difference of less than 5%.IC50 of DPPH and O-2·were 0.974 mg / m L and 1.342 mg / m L,respectively.Antioxidant activity of sample polysaccharides was weaker than those of vitamin C.Conclusion: The optimum enzymatic extraction technology of the polysaccharides from Camellia japonica L.by RSM was convenient and feasible,and the extracted polysaccharides had good antioxidant activity.