猪胰脏中胰酶的制备新工艺技术研究

Studies on the New Preparation Technology of Trypsin from Porcine Pancreas

以猪胰脏为原料,采用单因素试验研究胰酶制备的不同工艺路线、不同提取溶剂、提取激活温度、时间、pH和沉淀剂种类对三酶活力和收率及其他品质的影响,并通过正交试验优化得胰酶制备的工艺条件为:提取溶剂为25%乙醇(原料量的2倍v,.m-2),保护剂为1.0%氯化钠和1.0%甘油,激活剂为0.2%CaCl2和1.0%胰酶原粉;提取激活温度为20±1℃、pH 5.5、时间6 h;沉淀剂为0.2%壳聚糖、温度4℃或常温,预冷丙酮脱脂,40℃下真空干燥后,粉碎即得产品。在该工艺条件下制备的胰酶制品,其胰蛋白酶、胰淀粉酶、胰脂肪酶的酶活力分别达3 892、45 551、27 239 u.g-1,为中国药典所规定的三酶最低酶活力的6倍以上,平均收率12.63%,同时胰酶制品的其它指标也符合药典要求。

Using the pig pancreas as raw material,single-factor test of pancreatic enzyme preparation was used to research the effects of different process routes,different extraction solvent,extraction and activation temperature,time,pH,precipitant type on the three kinds of enzyme activity.Yield and other quality of pancreatin,and pancreatin preparation conditions optimized by orthogonal test were as follows： extraction solvent was 25% ethanol（the amount of it was 2 times of raw materials,v/m）,enzyme protectants were 1.0% sodium chloride and 1.0% glycerol,enzyme activator were 0.2 % CaCl2 and 1.0% original trypsin powder;extracting or activating temperature was 20±1 ℃,pH was 5.5,time was 6 h;precipitating agent was 0.2% chitosan,precipitating temperature was 4 ℃ or room temperature,a degreasing agent was pre-cooling acetone,vacuum drying temperature was 40 ℃,and the product obtained was comminuted.The pancreatin products prepared under the conditions of process optimization,where activity of trypsin,pancreatic amylase,pancreatic lipase were up to 3 892,45 551,27 239 u·g-1,respectively.Furthermore,the average yield of pancreatin was 12.63%,it＇s enzyme activity was more than 6 times that of the minimum enzyme activity to be required by the Chinese Pharmacopoeia,while other indicators of pancreatin products were in accordance with the requirements of the Chinese Pharmacopoeia.