利用红曲霉与乳酸菌混合发酵藜麦制备降压肽,以期得到一种藜麦蛋白源血管紧张素转化酶(angiotensin-I converting enzyme,ACE)抑制肽。从福建古田等地采集到的红曲样品中分离得到6株紫色红曲霉(Monascus purpureus)并结合实验室保存的菌种,对蛋白酶酶活力、蛋白水解度、粗多肽得率3个指标进行考察,筛选出适合发酵的菌种,并通过混菌验证试验确定戊糖片球菌(Pediococcus pentosaceus)22227和紫色红曲霉(Monascus purpureus)4422为最佳发酵组合,ACE抑制率高达84.21%。通过单因素试验优化发酵条件,当培养温度为28 ℃,P. pentosaceus 22227和M. purpureus 4422菌种比例为为2∶1,发酵7 d,制备的降压肽ACE抑制率最高,达到83.13%。将所得发酵上清液依次经过透析、葡聚糖(Sephadex G-25)凝胶过滤色谱、反相高效液相色谱(reversed phase-HPLC,RP-HPLC)3个步骤提纯,最终使得降压肽的ACE抑制率提高到了89.75%;氨基酸成分分析表明,所得降压肽含有12种氨基酸,其中包括丙氨酸、缬氨酸和甲硫氨酸等疏水性氨基酸,且占到氨基酸总量的50%以上,这可能和降压效果有关。该降压肽在低温、弱酸性条件下和体外连续消化试验过程中也能保持良好的活性。
[1] ZHENG Y J,WANG X,ZHUANG Y L,et al.Isolation of novel ACE-inhibitory and antioxidant peptides from quinoa bran albumin assisted with an in silico approach:Characterization,in vivo antihypertension,and molecular docking[J].Molecules,2019,24(24):4 562.
[2] DASKAYA-DIKMEN C,YUCETEPE A,KARBANCIOGLU-GULER F,et al.Angiotensin-I-converting enzyme (ACE)-inhibitory peptides from plants[J].Nutrients,2017,9(4):316.
[3] GUO H M,HAO Y Q,RICHEL A,et al.Antihypertensive effect of quinoa protein under simulated gastrointestinal digestion and peptide characterization[J].Journal of the Science of Food and Agriculture,2020,100(15):5 569-5 576.
[4] ALUKO R E,MONU E.Functional and bioactive properties of quinoa seed protein hydrolysates[J].Journal of Food Science,2003,68(4):1 254-1 258.
[5] RAVISANKAR S,CAMPOS G,CHIRINOS R,et al.Quinoa (Chenopodium quinoa) peptides protect human umbilical vein endothelial cells (HUVEC) against risk markers for cardiovascular disease (CVD)[C].The FASEB Journal,2015,291.
[6] 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.
LI J,CHEN D G,MA L.Optimization of fermentation conditions for preparation of ACE inhibitory peptides from soybean meal by Bacillus subtilis using response surface methodology[J].China Oils and Fats,2018,43(6):31-35.
[7] CHEN L,ZHANG Q H,JI Z,et al.Production and fermentation characteristics of angiotensin-I-converting enzyme inhibitory peptides of goat milk fermented by a novel wild Lactobacillus plantarum 69[J].LWT,2018,91:532-540.
[8] ZAREIAN M,OSKOUEIAN E,MAJDINASAB M,et al.Production of GABA-enriched idli with ACE inhibitory and antioxidant properties using Aspergillus oryzae:The antihypertensive effects in spontaneously hypertensive rats[J].Food & Function,2020,11(5):4 304-4 313.
[9] 范吉釴, 柯义强,刘红海,等.发酵法制备生物活性肽的研究进展[J].安徽农学通报,2020,26(23):19-23.
FAN J Y,KE Y Q,LIU H H,et al.Research progress in the preparation of bioactive peptides by fermentation[J].Anhui Agricultural Science Bulletin,2020,26(23):19-23.
[10] OBAROAKPO J U,LIU L,ZHANG S W,et al.α-Glucosidase and ACE dual inhibitory protein hydrolysates and peptide fractions of sprouted quinoa yoghurt beverages inoculated with Lactobacillus casei[J].Food Chemistry,2019,299:124 985.
[11] 李善仁. 混菌发酵豆粕制备大豆肽的研究[D].福州:福建农林大学,2009.
LI S R.Study on soybean peptides from soybean meal fermented by mixed strains[D].Fuzhou:Fujian Agriculture and Forestry University,2009.
[12] 何泽贺, 肖宇,刘建朝,等.大鳞副泥鳅蛋白制备ACE抑制肽的酶解方式研究[J].食品研究与开发,2021,42(7):134-140.
HE Z H,XIAO Y,LIU J C,et al.Study on enzymatic hydrolysis of ACE inhibitory peptide from loach (Paramisgurnus dablianus) protein[J].Food Research and Development,2021,42(7):134-140.
[13] 王銮, 包怡红,康宁.混菌固态发酵榛仁粕制备降血压肽工艺优化研究[J].中国粮油学报,2018,33(12):35-41.
WANG L,BAO H Y,KANG N.Optimization of preparation of antihypertensive peptide from hazelnut meal by solid-state fermentation with mixed bacteria[J].Journal of The Chinese Cereals and Oils Association,2018,33(12):35-41.
[14] 刘子毅. 玉米黄粉ACE抑制肽的制备及其活性研究[D].北京:中国农业科学院,2019.
LIU Z Y.Study on preparation and activity of ACE inhibitory peptide from corn gluten meal[D].Beijing:Chinese Academy of Agricultural Sciences,2019.
[15] 周亚丽. 魔芋ACE抑制肽分离纯化及体外降血压活性研究[D].西安:陕西科技大学,2017.
ZHOU Y L.Study on separation and purification of ACE inhibitory peptides from konjac and its antihypertensive activity in vitro[D].Xi’an:Shaanxi University of Science & Technology,2017.
[16] CHEN Y Y,GAO X,WEI Y X,et al.Isolation,purification and the anti-hypertensive effect of a novel angiotensin I-converting enzyme (ACE) inhibitory peptide from Ruditapes philippinarum fermented with Bacillus natto[J].Food & Function,2018,9(10):5 230-5 237.
[17] 罗鹏. 葵花籽ACE抑制肽的分离纯化、结构分析与稳态化研究[D].武汉:华中农业大学,2018.
LUO P.Separation and purification,structural analysis and stabilization of ACE inhibitory peptides derived from sunflower seeds[D].Wuhan:Huazhong Agricultural University,2018.
[18] 陈秋銮, 陈雪芹,马倩,等.酶解法制备牡丹籽ACE抑制肽及其稳定性[J].食品工业科技,2020,41(19):149-156.
CHEN Q L,CHEN X Q,MA Q,et al.Preparation and stability of ACE inhibitory peptides from peony seed meal by enzymatic hydrolysis[J].Science and Technology of Food Industry,2020,41(19):149-156.
[19] 杨高继, 刘欢欢,马艳,等.糙米中蜡样芽孢杆菌的分离鉴定及其毒力基因与药敏性检测[J].现代食品科技,2020,36(10):290-299.
YANG G J,LIU H H,MA Y,et al.Isolation,identification of Bacillus cereus from brown rice and its virulence genes detection and antibiotic susceptibility[J].Modern Food Science and Technology,2020,36(10):290-299.
[20] WANG L J.,SAITO M,TATSUMI E,et al.Antioxidative and angiotensin I-converting enzyme inhibitory activities of sufu (fermented tofu) extracts[J].Japan Agricultural Research Quarterly,2003,37(2):129-132.
[21] 李响, 刘畅,吴非.微生物发酵法制备大豆ACE抑制肽菌种的筛选[J].食品科学,2013,34(1):185-188.
LI X,LIU C,WU F.Screening of optimal strain for microbial preparation of ACE inhibitory peptides from soybean protein isolate[J].Food Science,2013,34(1):185-188.
[22] 谯飞. 大米蛋白ACE抑制肽制备及其膜高效分离技术的研究[D].哈尔滨:哈尔滨工业大学,2016.
QIAO F.The preparation of rice protein ACE inhibitory peptides and the research on its effective separation by membrane technology[D].Harbin:Harbin Institute of Technology,2016.
[23] 李勇. 蚕蛹ACE抑制肽的制备及其ACE抑制活性的研究[D].重庆:西南大学,2012.
LI Y.Study on pupa ACE inhibitory peptides preparation and its ACE inhibitory activity[D].Chongqing:Southwest University,2012.
[24] MIGUEL M,ALONSO M J,SALAICES M,et al.Antihypertensive,ACE-inhibitory and vasodilator properties of an egg white hydrolysate:Effect of a simulated intestinal digestion[J].Food Chemistry,2007,104(1):163-168.
[25] DOU B X,LIU Y,LIU Y M,et al.Isolation and characterization of angiotensin I converting enzyme (ACE) inhibitory peptides from rice bran proteins and evaluation of activity and stability[J].Pakistan Journal of Zoology,2020,52(4):1 397-1 405.
