胃蛋白酶法提取猪皮胶原蛋白的条件优化

Optimizing processing of collagen extraction from pigskin by pepsin

目的采用胃蛋白酶法从新鲜猪皮中提取胶原蛋白,以胶原蛋白提取收率及机械强度为考察指标,确定最佳酶解条件。方法胃蛋白酶解提取猪皮中胶原蛋白,扫描电镜观察胶原蛋白的表面形貌,通过傅里叶红外光谱(FTIR)法及拉力试验机分析胶原蛋白的化学结构及机械强度,通过测定胶原中羟脯氨酸的含量测定胶原提取收率,设计正交试验,探索最佳酶解条件。结果成功从新鲜猪皮中提取胶原蛋白,红外光谱测试显示提取产物具有三股螺旋结构,在酶解温度20℃,酶浓度2%,酶溶液pH 2.0,酶解时间24 h时提取胶原蛋白质量最优,测得胶原得率(83.7±2.8)%,断裂强度为(0.034±0.003)N.mm-2。结论胃蛋白酶可以在较温和的条件下高效率地将胶原蛋白从猪皮组织中提取出来,提取工艺高效简洁。

Objective To extract and purify collagen from the fresh pigskin, and make sure the optimum preparation conditions according to yield and tensile strength for collagen by enzyme hydrolysis extraction. Methods Collagen was extract from the pigskin with pepsin. The surface morphous of collagen was observed by scanning electron microscope （SEM） , the chemcial constitution and tensile strength of collagen protein were analyzed by Fourier transform infrared （FTIR） and tensile testing machine. The yield of the collagen were determine by testing the hydroxyproline content in the collagen. Enzymatic con- dition was optimized by the orthogonal design with collagen yield and tensile strength as indexes. Results Collagen was ex- tracted from the pig skin successful, it forms a triple helix conformation by FTIR. The optimum extraction condition was ob- tained as follows ： enzyme content 2% , pH 2.0, enzymatic hydrolyzed for 24 hours at 20 ℃. The yield of collagen was （ 83.7 ±2. 8 ） %, breaking strength was （ 0. 034 ± 0. 003 ） N·mm - 2. Conclusion This method that extract and purify collagen from the pigskin by pepsin is high efficiency and terse.