1. 吉林农业大学中药材学院,吉林,长春,130118
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李慧萍, 李梁, 范宁, 等. 东北林蛙体胶原蛋白酶法制备工艺[J]. 上海中医药杂志, 2015,49(12):78-83.
LI Hui-ping, LI Liang, FAN Ning, et al. Preparation technology of collagen from Rana dybowskii using pepsin[J]. Shanghai Journal of Traditional Chinese Medicine, 2015,49(12):78-83.
目的:探索胃蛋白酶法制备东北林蛙体胶原蛋白的最佳工艺条件,并对提取得到的胶原蛋白的纯度及其氨基酸含量进行测定。 方法:采用单因素及正交试验,以提取率为指标,优化胃蛋白酶提取东北林蛙蛙体胶原蛋白的工艺条件。通过紫外光谱(UV)分析确定东北林蛙体胃蛋白酶溶性胶原蛋白(PSC)的类型,反相高效液相色谱法(RP-HPLC)测定PSC中氨基酸的含量。 结果:PSC最佳提取工艺为温度37 ℃,E/S 20%,料液比1∶20,时间12 h;在此条件下,胶原蛋白提取率达到88.6%。经紫外光谱(UV)分析,PSC在234 nm处有最强吸收峰,反相高效液相色谱法(RP-HPLC)测定发现,PSC氨基酸含量丰富,包含人体所需的8种必需氨基酸,其中甘氨酸(Gly)、脯氨酸(Pro)、亮氨酸(Leu)和丙氨酸(Ala)等氨基酸含量较高,符合Ⅰ型胶原蛋白特征。 结论:优化后的制备方法提取率高、重复性好,可为PSC的工业化生产提供参考。
Objective:To explore the optimum technological conditions for preparing collagen from brown frog(Rana Dybowskii) body by pepsin,and to determine the purity of collagen and contents of amino acids. MethodsBy determining extraction rate,single-factor trials and orthogonal experiments were used to optimize technological conditions for preparing collagen from the body of this brown frog with pepsin. The type of pepsin-solubilized collagen(PSC) was determined by ultraviolet and visible spectrophotometer(UV),and the contents of amino acids in PSC were determined by using reversed phase high performance liquid chromatography(RP-HPLC); ResultsThe optimum technological conditions of PSC were that 37 ℃ for temperature,20% for E/S,1∶20 for the ratio of material to solvent and 12h for time. Under these conditions,the extraction rate of PSC was up to 88.6%. The PSC had the strongest absorption peak at 234 nm through UV analysis. Results: of RP-HPLC analysis showed that PSC contained abundant of amino acids,especially 8 essential amino acids needed by human body,in which Gly,Pro,Leu and Ala had higher contents than the others,which matched with the features of collagen Ⅰ. Conclusion:The optimized preparation method had high extraction rate and good repeatability, thus could provide references for technological production of PSC.
东北林蛙体胃蛋白酶胶原蛋白制备工艺必需氨基酸
brown frog bodypepsincollagenpreparation technologyessential amino acid
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