食品科学

• 生物工程 • 上一篇    下一篇

干酪乳清酶解产物抗氧化肽的分离和纯化

霍建新1,原慧艳1,王 燕1,王 演1,白彩艳1,赵文博2,*   

  1. 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
  • 出版日期:2015-07-15 发布日期:2015-07-08

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

HUO Jianxin1, YUAN Huiyan1, WANG Yan1, WANG Yan1, BAI Caiyan1, ZHAO Wenbo2,*   

  1. 1. Jinzhong University, Jinzhong 030600, China; 2. School of Food Engineering and Biological Technology,
    Tianjin University of Science and Technology, Tianjin 300457, China
  • Online:2015-07-15 Published:2015-07-08

摘要:

干酪乳清Alcalase 2.4L酶解的最优条件,即酶解时间为2 h、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-sodiumdodecyl 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4 000~6 000 D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。

关键词: 干酪乳清, 超滤, 碱性蛋白酶, 抗氧化肽

Abstract:

Cheese whey hydrolysates were obtained by enzymatic hydrolysis with an alkaline protease “alcalase” for 2 h
at 50 ℃ and an initial pH 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 pH 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.

Key words: cheese whey, ultrafiltration, Alaclase 2.4L, antioxidant peptide

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