高压酶解法提取硫酸皮肤素工艺优化及其抗氧化活性分析

以猪肠粘膜提取肝素过程中透过液为原料，优化硫酸皮肤素提取工艺，探讨硫酸皮肤素的抗氧化能力。以硫酸皮肤素浓度为检测指标，筛选酶解最佳用酶，在单因素基础上通过Box-Behnken响应曲面试验优化高压酶解工艺，筛选最优大孔树脂，优化动态吸附洗脱条件纯化硫酸皮肤素，探究体外抗氧化活性。结果显示:在最佳酶解条件温度56 ℃，pH9.0，时间8.4 h，压力135 MPa下实测酶解硫酸皮肤素浓度为13.05 ng/mL，接近模型值；D312树脂以1.5 BV/h流速上样，4% NaOH 1.5 BV/h流速洗脱纯化效果最好，高效凝胶渗透色谱测定纯度达73.32%；硫酸皮肤素抗氧化能力与浓度呈正相关，DPPH自由基在2.4 mg/mL时清除率为36.29%，表现出一定清除能力，对于·OH 2.4 mg/mL时清除能力较强为68.2%，总还原力于2.4 mg/mL时为0.22。该研究为分离纯化硫酸皮肤素及其在抗氧化功能方面开发利用提供理论依据。

Optimization of High-pressure Enzymatic Hydrolysis for Extraction of Dermatan Sulfate and Analysis of Its Antioxidant Activity

Taking permeate liquid in the process of extracting heparin from porcine intestinal mucosa as raw material, the extraction process of dermatan sulfate was optimized to explore the antioxidant capacity of dermatan sulfate. Taking the concentration of dermatan sulfate as the detection indicator, screening the best enzyme for enzymatic hydrolysis, on the basis of single factors, the high-pressure enzymatic hydrolysis process was optimized by Box-Behnken response surface test, the best macroporous resins were screened, the dynamic adsorption and elution conditions were optimized to purify dermatan sulfate, and investigating antioxidant activity in vitro. The results showed that: Under the optimal enzymatic hydrolysis conditions, temperature 56 ℃, pH9.0, time 8.4 h, and pressure 135 MPa, the measured concentration of dermatan sulfate was 13.05 ng/mL, which was close to the model value. D312 resin at 1.5 BV/h flow rate, 4% NaOH 1.5 BV/h flow rate elution and purification effect was the best, and high-performance gel permeation chromatography determination purity could reach 73.32%. The antioxidant capacity of dermatan sulfate was positively correlated with the concentration, the scavenging rate of DPPH free radical was 36.29% at 2.4 mg/mL, showing a certain scavenging ability. For ·OH, the scavenging ability was stronger at 2.4 mg/mL, which was 68.2%, and the total reducing power was 0.22 at 2.4 mg/mL. This study would provide a theoretical basis for the separation and purification of dermatan sulfate and its development and utilization in terms of antioxidant function.