以豆制品加工废水豆清液为原料,通过乳酸菌发酵制备大豆活性多肽并对其抗氧化活性进行研究。以多肽产率为评价指标,在单因素的基础上,采用响应面软件对乳酸菌发酵豆清液制备多肽的工艺建立数学模型;并对最优条件下的多肽清除羟基自由基(·OH)、2,2-连氮基-双-(3-乙基苯并二氢噻唑啉-6-磺酸)[2,2′-Azinobis-(3-ethylbenzthiazoline-6-sulphonate),ABTS]自由基、1,1-二苯基-2-苦肼基自由基(1,1-diphenyl-2-picrylhydrazyl free radical,DPPH)自由基的EC50进行测定。方差分析结果表明,影响多肽产率大小顺序为菌种添加量>发酵温度>初始pH>发酵时间;回归模型分析表明,乳酸菌最佳发酵条件为:菌种添加量为4.71%(4.71×108 CFU/100 mL)、初始pH为6.44、发酵温度为40.21 ℃、发酵时间为11.66 h,预测多肽产率为(56.97±0.27)%;结合试验具体情况调整发酵工艺参数并进行验证,实测多肽产率为(57.33±0.32)%,与预测值差异不显著(P>0.05);体外抗氧化结果表明,发酵后的豆清液多肽对·OH、ABTS、DPPH自由基的清除率的EC50分别为17.84、1.14、7.61 mg/mL,较发酵前分别提高了1.2、4.75和2.16倍。为豆清液高值化开发利用提供试验依据。
关键词：
乳酸菌; 发酵; 豆清液; 多肽; 抗氧化
Using the supernatant from the waste-water produced in soybean product processing as raw material, the soybean active peptide was prepared by lactic acid bacteria fermentation and its antioxidant activity was studied. Taking peptide yield as evaluation index, based on the results from single factor test, a response surface software was used to establish a mathematical model for the process of preparing peptide from fermented soybean supernatant by lactic acid bacteria. The EC50 for peptide scavenging of ·OH, ABTS and DPPH radical activity was determined. ANOVA results showed that the order of affecting the peptide yield was: bacteria addition amount> fermentation temperature> initial pH> fermentation time. Response surface model regression analysis showed that the optimal fermentation conditions were as follows: inoculation 4.71% (4.71×108 CFU/100 mL), initial pH 6.44, 40.21 ℃, and fermented for 11.66 h. The predicted peptide yield was (56.97±0.27)%. Adjusting the fermentation parameters in combination with the specific conditions of the experiment, then the verified peptide yield was (57.33 ± 0.32) %, which was not significantly different from the predicted value (P> 0.05). The results of in vitro antioxidant tests showed that the EC50 of the ·OH, ABTS, and DPPH free radical scavenging by the peptide was 17.84, 1.14 and 7.61 mg/mL, respectively, which were 1.2, 4.75 and 2.16 times higher than the value before fermentation. The research provided experimental basis for the high value development and utilization of soybean supernatant.
[1] 刘朝, 乔建军, 朱宏吉. 乳酸菌肽聚糖的研究进展[J]. 微生物学通报, 2016, 43(1): 188-197.
[2] 张婷婷, 翟齐啸, 金星,等. 具有拮抗空肠弯曲杆菌能力鸡源乳酸菌的筛选及特性[J]. 微生物学通报, 2017, 44(1): 118-125.
[3] 廖雪义, 郭丽琼, 邱灵燕, 等. 芒果源高降胆固醇乳酸菌的特性及其作用机制[J]. 中国食品学报, 2016,16(5):10-18.
[4] 靳志敏, 王柏辉, 罗玉龙, 等. 乳酸菌对小鼠脂质代谢及肠道结构的影响[J]. 中国食品学报, 2016, 16(6):30-36.
[5] 张庆峰, 吴祖芳, 张鑫, 等. 复合乳酸菌接种发酵对浙东腌冬瓜发酵品质的影响[J]. 核农学报, 2017, 31(3): 500-507.
[6] 李彤, 彭珍, 熊涛. 乳酸菌发酵对复合豆乳饮料营养成分、香气成分及抗氧化活性的影响[J]. 食品与发酵工业, 2018, 44(4): 111-118.
[7] 方靖, 陈中, 林伟锋, 等. 乳酸菌发酵剂对直接发酵法生产面包贮藏特性的影响[J]. 食品与发酵工业, 2014, 40(5): 101-105.
[8] KUNJI E R S, MIERAU I, HAGTING A, et al. The proteotytic systems of lactic acid bacteria [J]. Antonie Van Leeuwenhoek, 1996, 70(2-4): 187-221.
[9] 李景, 陈定刚, 马龙. 响应面优化枯草芽孢杆菌发酵豆粕制备ACE抑制肽条件[J]. 中国油脂, 2018, 43(6): 31-35.
[10] GIOVANNA B, GRAZIANA-MARIA S, DONATELL R, et al. ACE-inhibitory activity of enzymatic protein hydrolysates from lupin and other legumes[J]. Food Chemistry, 2014, 145(7): 34-40.
[11] SINGH B P, VIJ S, HATJ S. Functional significance of bioactive peptides derived from soybean[J]. Peptides, 2014, 54(2): 171-179.
[12] PUCHALSKA P, MARINA M L, GARCÍA M C. Isolation and identification of antioxidant peptides from commercial soybean-based infant formulas[J]. Food Chemistry, 2014, 148(2): 147-154.
[13] 马福建, 高长城, 解迪, 等. 大豆11S球蛋白水解肽的制备及抗氧化研究[J]. 食品研究与开发, 2019, 40(11):89-93.
[14] ROBLES-RAMÍREZ M D C, EVA R G, REYES-DUARTE F J, et al. Effect of germinated soy protein on the growth of hela cervical cancer cells in female athymic mice [J]. Nutrition and Cancer, 2012, 64(8): 1 261-1 268.
[15] 孔彦卓, 尹乐斌, 雷志明, 等. 豆清液综合利用研究进展[J]. 现代农业科技, 2017(1):247-249.
[16] JACKSON C J C, DINI J P, LAVANDIER C, et al. Effects of processing on the content and composition of isoflavones during manufacturing of soy beverage and tofu[J]. Process Biochemistry, 2002, 37(10):1 117-1 123.
[17] XU L, LAMB K, LAYTON L, et al. A membrane-based process for recovering isoflavones from a waste stream of soy processing[J]. Food Research International, 2004, 37(9):867-874.
[18] 郭珺, 武爱莲, 闫敏, 等. 利用豆制品废水发酵枯萎病拮抗菌Pb-4条件研究[J]. 生物技术通报, 2017, 33(8):167-173.
[19] 谢灵来, 赵良忠, 尹乐斌, 等. 豆清发酵液点浆工艺研究[J]. 食品与机械, 2017, 33(1): 184-189;194.
[20] 李雪, 赵晨晨, 钱方, 等. 大豆乳清多肽饮品的开发[J]. 大连工业大学学报, 2017, 36(1): 10-13.
[21] 李明, 赵良忠, 王容, 等. 果汁型豆酸汤饮料稳定性动力学模型初探[J]. 食品安全质量检测学报, 2019,10(9):2 555-2 561.
[22] 鲁伟, 任国谱, 宋俊梅. 蛋白水解液中多肽含量的测定方法[J]. 食品科学, 2005, 26(7): 169-171.
[23] DOU J, MENG Y, LIU L, et al. Purification, characterization and antioxidant activities of polysaccharides from thinned-young apple[J]. International Journal of Biological Macromolecules, 2015, 72:31-40.
[24] YILMAZ-ERSAN L, QZCAN T, AKPINAR-BAYIZIT A, et al. Comparison of antioxidant capacity of cow and ewe milk kefirs[J]. Journal of Dairy Science, 2018, 101(5):3 788-3 798.
[25] MOYO B, OYEDEMI S, MASIKA P J, et al. Polyphenolic content and antioxidant properties of Moringa oleifera leaf extracts and enzymatic activity of liver from goats supplemented with Moringa oleifera leaves/sunflower seed cake [J]. Meat Science, 2012, 91(4): 441-447.
[26] 陈燕飞. pH对微生物的影响[J]. 太原师范学院学报(自然科学版), 2009, 8(3): 121-124.
[27] 程旺开. 产蛋白酶乳酸菌的筛选及发酵豆奶产酶条件优化[J]. 芜湖职业技术学院学报, 2018, 77(1): 1-5.
[28] FIELDS P A. Temperature | proteins and temperature [J]. Encyclopedia of Fish Physiology, 2011: 1 703-1 708.
[29] 范宇. 乳酸菌对乳蛋白的降解作用及其应用研究[D]. 大连: 大连工业大学, 2009.
