Carrillo Wilman, Lucio Araceli, Gaibor Juan, Morales Dayana, Vásquez Grace
1 Research Department, Faculty of Health and Human Sciences, Bolivar State University , Guaranda, Ecuador .
2 Laboratory of Functional Foods, Faculty of Foods Science and Engineering, Technical University of Ambato , Ambato, Ecuador .
J Med Food. 2018 Aug;21(8):808-818. doi: 10.1089/jmf.2017.0134. Epub 2018 Apr 26.
Native and heated hen egg white lysozyme (HEWL) hydrolysates were isolated by hydrolysis with pepsin at pH 2.0 in situ in a cation exchange membrane to isolate and identify antibacterial peptides of the HEWL hydrolysates. Native and heated HEWL was partially hydrolyzed with pepsin at pH 2.0. The fractions were eluted with 5 M ammonia to identify 23 antibacterial peptides using a tandem mass spectrometry. Then, these fractions were eluted with a solution of NaCl 1 M, and seven positively charged peptides f(23-28) YSLGNW, f(122-129) AWIRGCRL, f(123-129) WIRGCRL, f(124-129) IRGCRL, f(82-96) ALLSSDITASVNCAK, f(103-129) VAWRNRCKGTDVQAWIRGCRL, and f(97-123) KIVSDGNGMNAWVAWRNRCKGT were identified using tandem mass spectrometry. Native HEWL hydrolysate presented an enzymatic activity of 23.0%, heated HEWL hydrolysate at pH 6.0 presented a residual enzymatic activity of 22.0%, and heated HEWL hydrolysate at pH 7.0 presented an enzymatic activity of 21.33%. Native and heated HEWL hydrolysate presented antibacterial activity against Escherichia coli and Staphylococcus carnosus. Native HEWL hydrolysate presented a higher enzymatic activity than heated HEWL hydrolysates.
通过在阳离子交换膜中于pH 2.0条件下用胃蛋白酶原位水解，分离天然和加热的鸡蛋清溶菌酶（HEWL）水解产物，以分离和鉴定HEWL水解产物中的抗菌肽。天然和加热的HEWL在pH 2.0条件下用胃蛋白酶进行部分水解。用5 M氨水洗脱各组分，使用串联质谱鉴定出23种抗菌肽。然后，用1 M NaCl溶液洗脱这些组分，使用串联质谱鉴定出7种带正电荷的肽，分别为f(23 - 28) YSLGNW、f(122 - 129) AWIRGCRL、f(123 - 129) WIRGCRL、f(124 - 129) IRGCRL、f(82 - 96) ALLSSDITASVNCAK、f(103 - 129) VAWRNRCKGTDVQAWIRGCRL和f(97 - 123) KIVSDGNGMNAWVAWRNRCKGT。天然HEWL水解产物的酶活性为23.0%，pH 6.0的加热HEWL水解产物的残余酶活性为2@0%，pH 7.0的加热HEWL水解产物的酶活性为21.33%。天然和加热的HEWL水解产物对大肠杆菌和肉葡萄球菌均具有抗菌活性。天然HEWL水解产物的酶活性高于加热的HEWL水解产物。
