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Domestic production of trypsin mainly pigs of trypsin, sheep trypsin or blank; sheep the trypsin case halal products, dedicated for the majority of Muslim consumers. In this study, the sheep pancreas to test material using organic solvent extraction sheep trypsin, preferably isopropanol as extraction solvent and precipitating agent; through single factor test and orthogonal design, the main influencing factors and process parameters studies to determine the trypsin extraction process and parameters, preferably a protective agent, activator and degreasing agent; reference to the People's Republic of China Pharmacopoeia determination of sheep trypsin vigor and physical and chemical indicators; sheep trypsin hydrolyzed casein dynamics. The results are as follows: 1. Process: the organic solvent method, frozen sheep pancreas as raw material, after the crushing of the pancreas, the activation, extraction, precipitation, degreasing, vacuum drying process such as obtained after the sheep pancreatin. 2. Extraction parameters: the preferred extractant concentration, activation time, activation temperature, activation of the pH optimum extraction conditions and parameter extraction temperature and extraction time. Selected main factors affect sheep trypsin extraction process and its parameters through single factor test, and further through L 18 (3 7 ) orthogonal test to trypsin vitality as a basis for screening out the best process parameters: 10% isopropyl alcohol concentration of the extractant, extraction temperature 15 ° C, extraction time 5h, activate pH6 activation temperature of 15 ° C, activation time of 5 h. 3. The preferred protective agent and activator: Screening maltose, starch, and sodium chloride as a protective agent Ca 2 - pancreatin powder activator, the sheep high dynamic trypsin. 4. The sedimentation process parameters: preferred precipitant concentration and precipitation temperature, precipitation precipitation conditions and parameters of the optimal pH and standing time. Elected by single factor experiment of the main factors affecting the precipitation of the trypsin and its parameter range, further through L 9 (3 4 ) orthogonal experiment, trypsin activity basis, filter out the optimal parameters: 60% of the precipitant concentration, a precipitation temperature 5 ° C, precipitation pH of 6, standing time 1h. 5 preferably acetone best degreasing agent, extracted at the above-optimum process sheep pancreatin, the determination of the vitality of the trypsin by trypsin vigor to 2654U / g, pancreatic amylase 13800U / g, pancreatic lipase enzyme activity 10400U / g, 11.2% of the sheep trypsin closing rate, fat residue to 17.1 mg of / g dry weight loss of 3.2%, are to meet the requirements of the Pharmacopoeia. 6 sheep tryptic casein kinetic characteristics: optimum temperature of 40 ° C, the optimum pH value of 8; by the thermal stability of the enzyme activity decreased gradually with time, the higher the temperature, the faster the trypsin inactivation , the worse the thermal stability; through the study of the pH stability, the enzyme activity decreased gradually with time, the optimum pH, trypsin inactivation fastest worse the pH stability; Ca 2 activation sheep trypsin, and improve the stability of trypsin, Na is no effect on the activity of trypsin, EDTA can inhibit trypsin activity. In summary, the use of isopropanol extract of sheep trypsin, the process is simple, high extraction efficiency, product activity, low cost, short production cycle. Improve the efficiency of production and raw material utilization, conserve resources and reduce pollution. Advanced bio-separation engineering Production of sheep trypsin, improve the comprehensive utilization of animal by-products recovery and preservation rate, biochemical pharmaceutical industry and the comprehensive utilization of animal by-products has a positive meaning.
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