菌酶协同下的低温压榨菜籽粕抗氧化肽制备工艺研究

Study on preparation process of antioxidant peptide from the lower temperature pressed rapeseed dregs with the synergy of microorganism and enzymes

以枯草芽孢杆菌、酿酒酵母、黒曲霉、米曲霉为供试菌株,碱性蛋白酶为供试酶,菜籽粕为发酵原料,采用液态发酵法制备具有抗氧化活性的菜籽多肽。以铁氰化钾还原法测定的菜籽多肽抗氧化活性吸光值为评价指标,进行料液比、p H、发酵温度、发酵时间、菌种组合、菌酶比单因素实验,再对料液比、p H、发酵温度和发酵时间4个因素做响应面实验,得到最佳发酵工艺参数:菌种组合为枯草芽孢杆菌、黑曲霉、米曲霉为(V/V/V=1∶1∶1),菌酶比为1∶1(V/V),接种量为10%(V/V),转速150 r/min,料液比为1∶20(g/m L),p H为8,发酵温度为36℃,发酵时间为36 h,此时菜籽多肽抗氧化能力最强,吸光值为1.26。此外,本文对最优工艺下制备的菜籽多肽的抗氧化活性也进行了测试,在设定的最大浓度0.7648 mg/m L处,多肽对超氧阴离子和DPPH自由基的清除率分别达到67.4%和76.78%,抗脂质过氧化活性达到61.5%,研究表明实验所制备的菜籽多肽具有较好的抗氧化活性。

In this paper,the preparation process of antioxidant peptide from the lower temperature pressed rapeseed dregs with the synergy of microorganism and enzymes was reported.Bacillus subtilis ,Saccharomyces cerevisiae,Aspergillus nige and A.oryzae were used for the rapeseed dregs liquid fermentation with alkaline protein enzyme. According to the rapeseed peptide＇s antioxidant activity tested by potassium ferricyanide reduction method, the following optimal operations were conducted that the single factor experiment of solid-liquid ratio, pH,fermentation temperature and time, strains combination and microorganism- enzyme ratio were separately done firstly. And then, the response surface test used the factors on solid- liquid ratio, pH, fermentation temperature and time was carried out as well.The best parameters combination was B.subtilis ,A.nige and A.oryzae strains（ V： V： V = 1 ： 1 ： 1 ） , microorganism- enzyme ratio was 1 ： 1 （V/V） , inoeulums size was 10% （V/V） , rotational speed was 150 r/min,solid-liquid ratio was 1：20 （g/mL）, pH 8, fermentation temperature was 36 ℃ and fermentation time was 36 h. And the peptide antioxidant activity reached the highest ahsorbance of 1.26 at this optimization condition.In addition,the oxidative stability of rapeseed peptides prepared used the optimal process was tested and the result showed that at the given maximal concentration of 0.7648 mg/mL,it had a good scavenging activity to superoxide anion and DPPH radical, which the scavenging rate was separately 67.4% and 76.78%. And its inhibition of lipid peroxidation reached 61.5% at the same concentration.The result showed the better antioxidant activity of the prepared rapeseed peptides.