Isolation,Purification And Structure Identification Of Pea Antitumor Peptides

Bioactive peptides,molecular weights less than 6000 Da,are beneficial to the living organism.Because of its biological activities and abundant structural types,peptides has become one of the most popular research objects and promising functional factors in the international food industry.In this study,the alkaline protease and trypsin were used to digest pea protein(PP),and the best conditions for APPH and TPPH were obtained through single factor analysis and orthogonal optimization.For APPH,the substrate concentration was 2%,the ratio of enzyme to substrate 18000 U/g,pH value was 8,the temperature was 55℃,the hydrolysis time was 4 hours,and under this condition,the degree of hydrolysis and the polypeptide content are respectively 36.73%and 60.46%.For TPPH,the substrate concentration was 6%,the ratio of enzyme to substrate 14000 U/g,pH value was 8.5,the temperature was 55℃,the hydrolysis time was 4 hours,and under this condition,the degree of hydrolysis and the polypeptide content are respectively 20.08%and 48.65%.To compared with the antiproliferative effect of PP,APPH,TPPH and 5-FU on human hepatocellular carcinoma cell line HepG2,human breast cancer cell line MCF-7,human gastric cancer cell line MGC803,human.lung cancer cell line A549 and human normal liver cell line L-02,MTT method was used.The results showed that neither the pea protein nor the pea peptides had no toxicity on human normal liver cells,and the inhibition of APPH on cancer cell lines,higher than TPPH,represented a time-dependent and dose-dependent dependent manner in the effective concentration of APPH.APPH-Ⅳ-2-1 and APPH-Ⅳ-2-3 with the high antitumor activity were obtained from APPH through the ultrafiltration,Sephadex G-15 and RP-HPLC.Then the LC-ESI-MS/MS were used.to analysis the peptide sequences.Finally,four peptides IPVNRPGQLQ,YGPTPVRDGFK,VTPGATDDQIMDGVRK and GQTPLFPR were considered to possess antitumor activity.