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Study On Preparation Of Immobilized Protease And Pine Nut Polypeptide Processing

Posted on:2010-09-21Degree:MasterType:Thesis
Country:ChinaCandidate:Y LiFull Text:PDF
GTID:2120360275967157Subject:Forest of plant resources
Abstract/Summary:PDF Full Text Request
Pinus koraiensis Sieb. et Zuee is a rare tree species of Xiao xing an ling in China. The pine nut is a high nutritional value of fruits, rich in protein, amino acids, fatty acids, vitamins, minerals and other substances physical activity has a wide application development potential.In this research, pine nuts were used as material, the use of modern analytical tools from the pine nut protein extraction technology, hydrolysis condition optimization; also research the best immobilized protease. The main results are as follows:1. Use ultrasound-assisted alkali extraction method from pine nut protein, optimum conditions were: Solid to liquid ratio (W/V) 1:24.61,pH10.0,extraction temperature48.3℃,ultrasonic time 100min. Pine nut protein extraction rate was 233.23 mg/g. Ultrasound-assisted extraction method can be effective in raising the pine nut protein extraction rate, shorten response cycle.2. Select Domestic the alkaline protease,neutral proteinase,the Alcalase2.4L alkalin proteinase and the Flavourzyme proteinase as a pine nut protein enzymolysis preparation enzyme, the enzyme effect pine nut protein research, in accordance with its hydrolysis of Alcalase2.4L alkalin proteinase identified as the best use of protease enzyme. Through the Response Surface Method to optimize the best hydrolysis conditions were: hydrolysis temperature 53℃, pH value 8.0, enzyme concentration 8800U/g pro, substrate concentration of 1.2%, hydrolysis time 50min. Under this best condition, the pine nut protein hydrolysis degree reached 43.15%. Its ratio of protease to protein has significant impact, followed by the pH value.3. To immobilize Alcalase2.4L alkalin proteinase with the cross-linking method, studied the process of immobilized Alcalase and the best conditions of immobilization, such as chitosan concentration 2.5%, glutaraldehyde concentration0.4% carrier cross-linking time4h, the weight of enzyme915U/g carrier, pH value 9.4, immobilization time8h, immobilization enzyme activity recovery rate was54.35%. Some characters of immobilized and free Alcalase2.4L, such as Michaelis constant, optimum temperature, pH, and thermal stability such as the nature were studied and compared. Repeated 6 times hydrolysis processes by immobilized Alcalase2.4L protease, the relative activity was still more than 60%. Illuminate immobilized Alcalase protease with the use of properties, can be used as industrial production.4. Studied immobilized Alcalase2.4L protease hydrolysis pine nut protein process. The optimal hydrolysis conditions were pH value9.0, temperature50℃, reaction time2h enzyme dosage0.2g/gsubstracte, substrate concentration 1.2%, hydrolysis degree is 25%. The use of immobilized Alcalase2.4L protease hydrolysis pine nut protein peptide preparation method is feasible.
Keywords/Search Tags:Pine nut protein, Enzymatic hydrolysis, Immobilize enzyme, Polypeptide
PDF Full Text Request
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