Bassan Juliana Cristina, de Souza Bezerra Thaís Milena, Peixoto Guilherme, da Cruz Clariana Zanutto Paulino, Galán Julián Paul Martínez, Vaz Aline Buda Dos Santos, Garrido Saulo Santesso, Filice Marco, Monti Rubens
Faculdade de Ciências Farmacêuticas, UNESP-Univ. Estadual Paulista, 14800-903, Departamento de Alimentos e Nutrição, Araraquara-SP, Brazil.
Instituto de Química, UNESP-Univ. Estadual Paulista, 14800-060, Departamento de Bioquímica e Tecnologia Química, Araraquara-SP, Brazil.
Materials (Basel). 2016 May 12;9(5):357. doi: 10.3390/ma9050357.
In this study, trypsin (Enzyme Comission 3.4.21.4) was immobilized in a low cost, lignocellulosic support (corn cob powder-CCP) with the goal of obtaining peptides with bioactive potential from cheese whey. The pretreated support was activated with glyoxyl groups, glutaraldehyde and IDA-glyoxyl. The immobilization yields of the derivatives were higher than 83%, and the retention of catalytic activity was higher than 74%. The trypsin-glyoxyl-CCP derivative was thermally stable at 65 °C, a value that was 1090-fold higher than that obtained with the free enzyme. The trypsin-IDA-glyoxyl-CCP and trypsin-glutaraldehyde-CCP derivatives had thermal stabilities that were 883- and five-fold higher, respectively, then those obtained with the free enzyme. In the batch experiments, trypsin-IDA-glyoxyl-CCP retained 91% of its activity and had a degree of hydrolysis of 12.49%, while the values for trypsin-glyoxyl-CCP were 87% and 15.46%, respectively. The stabilized derivative trypsin-glyoxyl-CCP was also tested in an upflow packed-bed reactor. The hydrodynamic characterization of this reactor was a plug flow pattern, and the kinetics of this system provided a relative activity of 3.04 ± 0.01 U·g and an average degree of hydrolysis of 23%, which were suitable for the production of potentially bioactive peptides.
在本研究中，将胰蛋白酶（酶委员会编号3.4.21.4）固定在低成本的木质纤维素载体（玉米芯粉-CCP）上，目的是从奶酪乳清中获得具有生物活性潜力的肽。对预处理后的载体用乙二醛基团、戊二醛和亚氨基二乙酸-乙二醛进行活化。衍生物的固定化产率高于83%，催化活性保留率高于74%。胰蛋白酶-乙二醛-CCP衍生物在65℃下具有热稳定性，该值比游离酶高1090倍。胰蛋白酶-亚氨基二乙酸-乙二醛-CCP和胰蛋白酶-戊二醛-CCP衍生物的热稳定性分别比游离酶高883倍和5倍。在分批实验中，胰蛋白酶-亚氨基二乙酸-乙二醛-CCP保留了91%的活性，水解度为12.49%，而胰蛋白酶-乙二醛-CCP的值分别为87%和15.46%。还在向上流动的填充床反应器中测试了稳定化衍生物胰蛋白酶-乙二醛-CCP。该反应器的流体动力学特征为活塞流模式，该系统的动力学提供了3.04±0.01 U·g的相对活性和23%的平均水解度，这适合于生产具有潜在生物活性的肽。
