Purification and Identification of Antioxidant Peptides from Enzymatic Hydrolysates of Tilapia (Oreochromis niloticus) Frame Protein 英文参考文献.docVIP

Molecules 2012, 17, 12836-12850; doi:10.3390/molecules171112836 OPEN ACCESS molecules ISSN 1420-3049 /journal/molecules Article Purification and Identification of Antioxidant Peptides from Enzymatic Hydrolysates of Tilapia (Oreochromis niloticus) Frame Protein Jian Fan, Jintang He, Yongliang Zhuang and Liping Sun * College of Chemistry and Engineering, Kunming University of Science and Technology, Kunming 650500, China; E-Mails: fj_3332000@ (J.F.); kmustslp@ (J.H.); kmylzhuang@ (Y.Z.) * Author to whom correspondence should be addressed; E-Mail: kmlpsun@; Tel.: +86-871-5920-302; Fax: +86-871-3801-114. Received: 29 September 2012; in revised form: 26 October 2012 / Accepted: 29 October 2012 / Published: 1 November 2012 Abstract: Tilapia frame protein was hydrolyzed by different proteases, including properase E, pepsin, trypsin, flavourzyme, neutrase, gc106 and papain, to obtain antioxidant peptides. The tilapia frame protein hydrolysate (TFPH) obtained by trypsin exhibited the highest degree of hydrolysis and antioxidant activity. Three series of peptides (TFPH1, TFPH 2 and TFPH 3) were obtained by ultrafiltration of TFPH through molecular weight cut-off membranes of 5, 3 and 1 kDa, respectively, and their IC50 values on scavenging 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical, superoxide anion radical (?O2), hydrogen peroxides (H2O2) and hydroxyl radical (?OH) activities were determined and compared with glutathione (GSH). The results showed that TFPH1 had the highest antioxidant activity. TFPH1 was further purified using ion exchange chromatography, gel filtration chromatography, and reversed phase high performance liquid chromatography (RP-HPLC). Finally, two antioxidant peptides were identified and the amino acid sequences were identified as Asp-Cys-Gly-Tyr (456.12 Da) and Asn-Tyr-Asp-Glu-Tyr (702.26 Da), respectively. The IC50 values of two peptides on hydro