响应面优化南极磷虾粉肽制备工艺及α-葡萄糖苷酶抑制活性分析

Optimization of preparation technology of Antarctic krill powder peptide by response surface methodology and analysis of its inhibitory activity onα-glucosidase

以南极磷虾(Euphausia superba)粉为原料,α-葡萄糖苷酶抑制率为指标,在确认最佳生物酶的基础上,通过单因素试验和响应面优化酶解工艺条件,并用超滤和G-25进行分离纯化,凝胶色谱法探究酶解液和多肽的分子量及其分布情况。结果表明:复合蛋白酶酶解效果最佳;最佳酶解条件为酶解时间5.9 h、料液比3.75∶1、酶添加量6.2%(原料);酶解液的相对分子质量集中分布在3000 u以下,占98.63%;经超滤C4(<3 ku)组分对α-葡萄糖苷酶抑制率IC_(50)为(14.89±2.15)mg/mL;G-25纯化C4-2组分对α-葡萄糖苷酶抑制率IC_(50)为(4.64±0.15)mg/mL。C4-2组分中<1000 u的多肽占97.6%,其中C4-2-1、C4-2-2、C4-2-3的分子量分别为597 u、335 u、246 u。

The best biological enzyme was selected according to the inhibition rate of α-glucosidase. Based on the single factor, optimization of enzymatic hydrolysis conditions could be obtained through the response surface. The enzymatic hydrolysate of the defatted antarctic krill powder was separated and purified by ultrafiltration and G-25. The molecular mass and distribution of enzymatic hydrolysate and peptides were studied by gel chromatography. The results showed that: The compound protease had the best enzymatic hydrolysis effect;The best enzymolysis conditions were enzymolysis time 5.9 h, material-liquid ratio 3.75∶1, enzyme addition amount 0.062 g/g(raw material);The molecular mass of the enzymatic hydrolysate was concentrated below 3 000 u, accounting for 98.63%;The C4 (<3 ku) component by ultrafiltration had the best inhibition rate of α-glucosidase [IC_(50) was (14.89±2.15) mg/mL];After purification by G-25, the C4-2 fractions (<3 ku) had higher α-glucosidase inhibitory than other fractions[IC_(50) was (4.64±0.14) mg/mL];In the C4-2 fraction, 97.6% of peptides were <1 000 u. The molecular weights of C4-2-1, C4-2-2, and C4-2-3 were 597 u, 335 u, and 246 u respectively. This research for the first time studied the method of preparing α-glucosidase inhibitory active peptide from Antarctic krill powder, which provides a basis for the comprehensive utilization of Antarctic krill resources.