响应面法优化鹅全骨硫酸软骨素的酶法提取工艺

Optimization of Enzymatic Extraction of Chondroitin Sulfate from Goose Bones by Response Surface Methodology

采用麻阳鹅鹅全骨为原料,以硫酸软骨素(chondroitin sulfate,CS)的提取率为指标,对运用碱提、复合酶解、醇沉法提取CS的工艺进行研究,确定使用胃蛋白酶进行酶解后,通过单因素试验和响应面优化试验,应用Box-Behnken实验设计建立二次多项式数学模型,进行响应面分析,确定胃蛋白酶酶解提取CS的最佳工艺参数。结果表明:碱提、复合酶解、醇沉法胃蛋白酶酶解提取鹅全骨CS的最佳工艺为加酶量1.55 g/L、酶解时间6.4 h、酶解温度42℃、pH值3.09,在此条件下CS的提取率为12.13%;通过高效液相色谱测定可知,提取物为CS和盐酸氨基葡萄糖。

The preparation of chondroitin sulfate（CS） from the bones of Mayang goose was investigated by alkali extraction, sequential enzymatic hydrolysis and ethanol precipitation. Pepsin was selected for the second hydrolysis step（the first hydrolysis step was carried out using trypsin） and hydrolysis conditions were optimized using one-factor-at-a-time method and response surface methodology. The extraction efficiency of CS was used as response. As a result, a polynomial quadratic regression model was developed using Box-Behnken design. The optimal hydrolysis conditions were determined as follows： enzyme concentration 1.55 g/L, hydrolysis time 6.4 h, temperature 42 ℃ and initial pH 3.09. Under these conditions, the extraction yield of CS was 12.13%. Chondroitin sulfate and glucosamine hydrochloride were identified in the extract by high performance liquid chromatography.