干酪乳清酶解产物抗氧化肽的分离和纯化  

Isolation and Purification of Antioxidant Peptide from Cheese Whey Hydrolysates Produced with Alkaline Protease

作　　者：霍建新[1] 原慧艳[1] 王燕[1] 王演[1] 白彩艳[1] 赵文博[2] 

机构地区：[1]晋中学院,山西晋中030600 [2]天津科技大学食品工程与生物技术学院,天津300457

出　　处：《食品科学》2015年第13期172-177,共6页Food Science

基　　金：山西省高等学校科技创新项目(2010127)

摘　　要：干酪乳清Alcalase 2.4L酶解的最优条件,即酶解时间为2h、pH 9.5、酶与底物比为4%、酶解温度为50℃。利用超滤、葡聚糖凝胶层析和三羟甲基氨基甘氨酸-十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（tricine-sodium dodecyl sulfate-polyacrylamide gelelectrophoresis,tricine SDS-PAGE）等方法从干酪乳清中提取抗氧化性蛋白肽。分别考察操作压力、料液温度、料液pH值、操作时间对超滤膜膜通量的影响。乳清酶解物超滤的最佳条件为：压力0.25 MPa、温度30℃、时间120 min、初始pH 9.0。分子质量4 000~6 000D肽的水解液脂质过氧化抑制率最高,达到47.28%。利用Sephadex G-50型葡聚糖凝胶进行纯化,将分离的组分进行Tricine-SDS-PAGE分析和脂质过氧化抑制率的测定。第34管洗脱液的脂质过氧化抑制率最高,分子质量范围为4 000~4 100 D。Cheese whey hydrolysates were obtained by enzymatic hydrolysis with an alkaline protease ＂alcalase＂ for 2 h at 50 ℃ and an initial p H of 9.5 with an [E]/[S] ratio of 4%.The antioxidant peptide was purified from cheese whey hydrolysates by ultrafiltration,polydextran gel chromatography and tricine-sodium dodecyl sulfate-polyacrylamide gelelectrophoresis（tricine SDS-PAGE）.The optimal operating pressure,hydrolysate temperature and p H and ultrafiltration time for the improved ultrafiltration membrane flux were found to be 0.25 MPa,30 ℃,9.0 and 120 min,respectively.The peptide with molecular weights ranging from 4 000 to 6 000 D had the highest inhibitory activity on lipid peroxidation,with an inhibitory rate of approximately 47.28%.After purification with Sephadex G-50,the separated fractions were determined by tricine SDS-PAGE and lipid peroxidation inhibition,respectively.Fraction 34 with molecular weight distribution ranging from 4 000 to 4 100 D had the highest inhibitory activity on lipid peroxidation.

关 键 词：干酪乳清 超滤 碱性蛋白酶 抗氧化肽 

分 类 号：TS252.9[轻工技术与工程—农产品加工及贮藏工程]