响应面法优化酶法制备猪血红蛋白抗氧化肽

以猪血红蛋白为原料,研究蛋白酶水解制备抗氧化肽的工艺。通过研究酶解时间、酶与底物比、酶种类对制备的抗氧化肽还原力和水解度影响,筛选出最佳蛋白酶及制备抗氧化肽的最佳工艺条件。结果表明:7种酶中,胃蛋白酶的水解度和还原力最佳。胰蛋白酶和胃蛋白酶复合水解比单酶水解的水解度提高了11.83%,还原力没有显著性差异。在此基础上设计单酶响应面,得到最佳还原力酶解条件为:酶解温度37.31℃、pH1.95、酶与底物比3526.74U/g。脱色条件为活性炭用量3%、pH4.0、温度70℃、脱色时间1h,粉末状活性炭的脱色率达到85.69%,蛋白质损失率20.32%。

Optimization of Enzymatic Preparation of Antioxidant Peptide from Porcine Hemoglobin

The enzymatic preparation of antioxidant peptides from porcine hemoglobin was studied. The effects of enzymatic hydrolysis time, enzyme/substrate ratio, and different proteases on the degree of hydrolysis （DH） and reducing power （RP） of antioxidant peptides were examined. Finally, the optimum enzyme and hydrolysis conditions for preparing antioxidant peptides were established. Pepsin was the best among seven proteases tested as evaluated by both DH and RP of antioxidant peptides. The combined use of pepsin with trypsin increased the DH by 11.83% compared with its single use without leading to a significant difference in the RP. Employing response surface methodology, the optimum hydrolysis conditions using pepsin alone were determined as follows： hydrolysis temperature of 37.31 ℃, initial pH 1.95 and enzyme/substrate ratio of 3526.74 U/g. The resulting hydrolysate was decolorized by adding 0.3% of activated charcoal and incubating at 70 ℃ for 1 h. Under this condition, the decolorization efficiency was 85.69% and the protein loss was about 20.32%.