Franco Indira, Castillo Eduardo, Pérez María-Dolores, Calvo Miguel, Sánchez Lourdes
Department of Food Technology, Veterinary Faculty, University of Zaragoza, Zaragoza, Spain.
Biosci Biotechnol Biochem. 2012;76(1):53-9. doi: 10.1271/bbb.110433. Epub 2012 Jan 7.
High pressure was applied to recombinant human lactoferrin obtained from rice (rhLF) and its effect was evaluated on the structure and activity of the protein. Treatments of 400, 500, and 650 MPa for 15 min (20 °C), were applied to rhLF at 2 mg/mL in three iron-saturation forms. The structural characteristics of the treated proteins were analyzed by differential scanning calorimetry (DSC) and by fluorometric analysis, and immunoreactivity by ELISA. Iron retention and binding properties and antibacterial activity against Escherichia coli O157:H7 were also studied. The results obtained indicate that the treatments at 400 and 500 MPa did not greatly modifiy the conformation of lactoferrin, meanwhile treatment at 650 MPa affected in different degrees the three forms of rhLF. With respect to antibacterial activity, only apo rhLF showed antibacterial activity against E. coli, activity that was maintained after treatment at 400 MPa, while holo and AsIs rhLF did not inhibit the growth of E. coli.
对从大米中获得的重组人乳铁蛋白（rhLF）施加高压，并评估其对该蛋白质结构和活性的影响。以三种铁饱和形式，对浓度为2 mg/mL的rhLF在20°C下分别施加400、500和650 MPa的压力15分钟。通过差示扫描量热法（DSC）、荧光分析对处理后的蛋白质结构特征进行分析，并通过酶联免疫吸附测定法（ELISA）分析免疫反应性。还研究了铁保留和结合特性以及对大肠杆菌O157:H7的抗菌活性。所得结果表明，400和500 MPa的处理对乳铁蛋白的构象影响不大，而650 MPa的处理对三种形式的rhLF有不同程度的影响。关于抗菌活性，只有脱铁rhLF对大肠杆菌具有抗菌活性，在400 MPa处理后该活性得以保持，而全铁和非铁全铁rhLF则不抑制大肠杆菌的生长。
