牦牛血抗氧化低聚肽制备工艺的优化及抗缺氧活性的初探
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摘要
为简化牦牛血抗氧化低聚肽的制备工艺并对其抗缺氧活性进行初探,实验采用菌酶联合发酵、超滤分级制备牦牛血抗氧化低聚肽,常压耐缺氧实验以及亚硝酸钠中毒存活实验评价其抗缺氧活性。结果表明:1 k Da超滤分级得到的牦牛血抗氧化低聚肽具有良好的体外抗氧化活性,·OH清除能力、DPPH·清除能力、总抗氧化力以及脂质过氧化抑制能力均极显著优于商品化大豆低聚肽(P <0. 01);不同剂量的牦牛血抗氧化低聚肽对小鼠常压缺氧存活时间以及亚硝酸钠中毒存活时间均有不同程度的延长,并呈现剂量依赖性。综上,牦牛血抗氧化低聚肽可作为一种天然抗氧化剂,且具有提高小鼠抗缺氧应激的能力。
In order to simplify the preparation process of yak blood antioxidant oligopeptides and study their antihypoxic activity,yak blood antioxidant oligopeptides were prepared by bacteria enzyme combined fermentation,5 kDa and 1 kDa ultrafiltration classification,the anti-hypoxia activities were evaluated by atmospheric hypoxia tolerance test and sodium nitrite poisoning survival test. The results showed that yak blood antioxidant oligopeptides had good antioxidant activity in vitro,·OH scavenging ability,DPPH· scavenging activity,total antioxidant ability,total reducing power and lipid peroxidation inhibition were significantly better than commercial soybean oligopeptides in vitro( P < 0. 01); different doses of yak blood antioxidant oligopeptides prolonged the survival time of mice under normal pressure hypoxia and sodium nitrite poisoning to varying degrees,and dose-dependent. In conclusion,yak blood antioxidant oligopeptides could be used as a natural antioxidant,and could improve the ability of mice to resist hypoxic stress,and the mechanism of anti-hypoxia could be further studied.
In order to simplify the preparation process of yak blood antioxidant oligopeptides and study their antihypoxic activity,yak blood antioxidant oligopeptides were prepared by bacteria enzyme combined fermentation,5 kDa and 1 kDa ultrafiltration classification,the anti-hypoxia activities were evaluated by atmospheric hypoxia tolerance test and sodium nitrite poisoning survival test. The results showed that yak blood antioxidant oligopeptides had good antioxidant activity in vitro,·OH scavenging ability,DPPH· scavenging activity,total antioxidant ability,total reducing power and lipid peroxidation inhibition were significantly better than commercial soybean oligopeptides in vitro( P < 0. 01); different doses of yak blood antioxidant oligopeptides prolonged the survival time of mice under normal pressure hypoxia and sodium nitrite poisoning to varying degrees,and dose-dependent. In conclusion,yak blood antioxidant oligopeptides could be used as a natural antioxidant,and could improve the ability of mice to resist hypoxic stress,and the mechanism of anti-hypoxia could be further studied.
引文
[1] SEGURA C M,CHEL G L,BETANCUR A D. Angiotensin-I converting enzyme inhibitory and antioxidant activities of peptide fractions extracted by ultrafiltration of cowpea Vigna unguiculata hydrolysates[J]. J Sci Food Agric,2010,90(14):2 512-2 518.
[2] CHAY P T B,MINE Y,JUNEJA L R,et al. Comparative composition and antioxidant activity of Peptide fractions obtained by ultrafiltration of egg yolk protein enzymatic hydrolysates[J]. Membranes(Basel),2011,1(3):149-161.
[3] WANG B,LI Z R,CHI C F,et al. Preparation and evaluation of antioxidant peptides from ethanol-soluble proteins hydrolysate of Sphyrna lewini muscle[J]. Peptides,2012,36(2):240-250.
[4] NAZEER R A,KUMAR N S,JAI G R. In vitro and in vivo studies on the antioxidant activity of fish peptide isolated from the croaker(Otolithes ruber)muscle protein hydrolysate[J]. Peptides,2012,35(2):261-268.
[5] IRVINE G B. High-performance size-exclusion chromatography of peptides[J]. J Biochem Biophys Methods,2003,56(1-3):233-242.
[6] CHEN H M,MURAMOTO K,YAMAUCHI F,et al. Antioxidative properties of histidine-containing peptides designed from peptide fragments found in the digests of a soybean protein[J]. J Agric Food Chem,1998,46(1):49-53.
[7] ZHANG Y,DUAN X,ZHUANG Y. Purification and characterization of novel antioxidant peptides from enzymatic hydrolysates of tilapia(Oreochromis niloticus)skin gelatin[J]. Peptides,2012,38(1):13-21.
[8] JIANG H,TONG T,SUN J,et al. Purification and characterization of antioxidative peptides from round scad(Decapterus maruadsi)muscle protein hydrolysate[J]. Food Chem,2014,154:158-163.
[9]杜昕.菌酶联合制备牦牛血抗氧化肽及其分离纯化的研究[D].雅安:四川农业大学,2016.
[10] CENTENARO G S,SALAS-MELLADO M,PIRES C,et al. Fractionation of protein hydrolysates of fish and chicken using membrane ultrafiltration:investigation of antioxidant activity[J]. Appl Biochem Biotechnol,2014,172(6):2 877-2 893.
[11] ROBLET C,DOYEN A,AMIOT J,et al. Enhancement of glucose uptake in muscular cell by soybean charged peptides isolated by electrodialysis with ultrafiltration membranes(EDUF):activation of the AMPK pathway[J]. Food Chem,2014,147:124-130.
[12] HE R,GIRGIH A T,ROZOY E,et al. Selective separation and concentration of antihypertensive peptides from rapeseed protein hydrolysate by electrodialysis with ultrafiltration membranes[J]. Food Chem,2016,197(Pt A):1 008-1 014.
[13]中华人民共和国卫生部.保健食品检验与评价技术规范(2003版)[S].北京:中国标准出版社,2003.
[14]魏稚萍,张才骏,杨荣珍,等.大通家牦牛及含野血牦牛血清亲血色蛋白多态性的比较研究[J].草食家畜,1996,1:22-23.
[15]张才骏,马凤莲,李黎,等.互助白牦牛血液蛋白质多态性的研究[J].青海畜牧兽医杂志,1996,26(3):9-11.
[16]魏稚萍.大通家牦牛及含野血牦牛血清运铁蛋白多态性的比较研究[J].甘肃畜牧兽医,1997,5:18-20.
[17]姚星辰,许博林,铁军,等.牦牛活性蛋白对小鼠抗缺氧抗疲劳能力及血糖的影响[J].时珍国医国药,2014,25(11):2 637-2 639.
[18]李冬利,于洪儒,杨菁,等.牛血活性肽对缺氧损伤心肌细胞的保护作用[J].中国新药杂志,2006,14:1 169-1 172.
[19]杜昕,李诚,肖岚,等.酶解牦牛血发酵液制备抗氧化肽工艺优化[J].核农学报,2017,31(2):325-333.
[20]杜昕,肖岚,李诚,等.枯草芽孢杆菌发酵牦牛血制备抗氧化肽工艺优化[J].食品与机械,2016,32(3):165-168; 205.
[21] TIAN F,LI B,JI B P,et al. Antioxidant and antimicrobial activities of consecutive extracts from Galla chinensis:The polarity affects the bioactivities[J]. Food Chemistry,2009,113(1):173-179.
[22] CHEN Naifu. Study on the extraction of flavonoid compound in pteridium aquilinum and its antioxidant property[J]. Food and Fermentation Industries,2003,29(11):63-67.
[23]刘昭明,黄翠姬,孟陆丽,等.核桃蛋白酶解物的抗氧化活性研究[J].食品与发酵工业,2009,35(1):58-61
[24]潘瑶,郑时莲,邹兴平,等.葡萄、芒果、草莓乙醇提取物抗氧化活性组分分析及其抗氧化相互作用[J].食品科学,2017,04:133-140.
[25]朱夕波.鲨鱼皮和猪皮胶原蛋白及其抗氧化活性酶解物的特性研究[D].上海:上海海洋大学,2008.
[26] YANG M,SHEN Q,LI L Q,et al. Phytochemical profiles,antioxidant activities of functional herb Abrus cantoniensis and Abrus mollis[J]. Food Chemistry,2015,177:304-312.
[27]张晖,唐文婷,王立,等.抗氧化肽的构效关系研究进展[J].食品与生物技术学报,2013,32(7):673-679.
[28]丁顺杰,罗金凤,丁晓雯,等.酶解缫丝蚕蛹蛋白抗氧化肽的分离与稳定性研究[J].食品科学,2015,36(3):35-40.
[29] CHI C F,CAO Z H,WANG B,et al. Antioxidant and functional properties of collagen hydrolysates from Spanish mackerel skin as influenced by average molecular weight[J]. Molecules,2014,19(8):11 211-11 230.
[30] SUETSUNA K,CHEN J R. Isolation and characterization of peptides with antioxidant activity derived from wheat gluten[J]. Food Science and Technology Research,2002,8(3):227-230.
[31]徐力,李相鲁,吴晓霞,等.一种新的玉米抗氧化肽的制备与结构表征[J].高等学校化学学报,2004,25(3):466-469.
[32] QIAN Z J,JUNG W K,KIM S K. Free radical scavenging activity of a novel antioxidative peptide purified from hydrolysate of bullfrog skin,Rana catesbeiana Shaw[J].Bioresource Technology,2008,99(6):1 690-1 698.
[33] SAMONINA G,ASHMARIN I,LYAPINA L. Glyproline peptide family:review on bioactivity and possible origins[J]. Pathophysiology,2002,8(4):229-234.
[34] BAILEY D M,BARTSCH P,KNAUTH M,et al. Emerging concepts in acute mountain sickness and high-altitude cerebral edema:from the molecular to the morphological[J]. Cell Mol Life Sci,2009,66:3 583-3 594.
[35] SCHREIBER M,TROJAN S. Protective effect of flavonoids and tocopherol in high altitude hypoxia in the rat:comparison with ascorbic acid[J]. Cesk Fysiol,1998,47:51-52.
[36] AL-HASHEM F H. Potential roles for vitamins E and C in combination in modulating exhaustive swimming and high altitude-associated lung injury in rats[J]. Saudi Med J,2012,33:367-374.
[37] FAN Pengcheng,MA Huiping,JING Linlin,et al. The antioxidative effect of a novel free radical scavenger 4'-hydroxyl-2-substituted phenylnitronyl nitroxide in acute highaltitude hypoxia mice. Biological&Pharmaceutical Bulletin,2013,36(6):917-924.
[38] MASTER S,GOTTSTEIN J,BLEI A T. Cerebral blood flow and the development of ammonia-induced brain edema in rats after portacaval anastomosis[J]. Hepatology,1999,30:876-880.
[2] CHAY P T B,MINE Y,JUNEJA L R,et al. Comparative composition and antioxidant activity of Peptide fractions obtained by ultrafiltration of egg yolk protein enzymatic hydrolysates[J]. Membranes(Basel),2011,1(3):149-161.
[3] WANG B,LI Z R,CHI C F,et al. Preparation and evaluation of antioxidant peptides from ethanol-soluble proteins hydrolysate of Sphyrna lewini muscle[J]. Peptides,2012,36(2):240-250.
[4] NAZEER R A,KUMAR N S,JAI G R. In vitro and in vivo studies on the antioxidant activity of fish peptide isolated from the croaker(Otolithes ruber)muscle protein hydrolysate[J]. Peptides,2012,35(2):261-268.
[5] IRVINE G B. High-performance size-exclusion chromatography of peptides[J]. J Biochem Biophys Methods,2003,56(1-3):233-242.
[6] CHEN H M,MURAMOTO K,YAMAUCHI F,et al. Antioxidative properties of histidine-containing peptides designed from peptide fragments found in the digests of a soybean protein[J]. J Agric Food Chem,1998,46(1):49-53.
[7] ZHANG Y,DUAN X,ZHUANG Y. Purification and characterization of novel antioxidant peptides from enzymatic hydrolysates of tilapia(Oreochromis niloticus)skin gelatin[J]. Peptides,2012,38(1):13-21.
[8] JIANG H,TONG T,SUN J,et al. Purification and characterization of antioxidative peptides from round scad(Decapterus maruadsi)muscle protein hydrolysate[J]. Food Chem,2014,154:158-163.
[9]杜昕.菌酶联合制备牦牛血抗氧化肽及其分离纯化的研究[D].雅安:四川农业大学,2016.
[10] CENTENARO G S,SALAS-MELLADO M,PIRES C,et al. Fractionation of protein hydrolysates of fish and chicken using membrane ultrafiltration:investigation of antioxidant activity[J]. Appl Biochem Biotechnol,2014,172(6):2 877-2 893.
[11] ROBLET C,DOYEN A,AMIOT J,et al. Enhancement of glucose uptake in muscular cell by soybean charged peptides isolated by electrodialysis with ultrafiltration membranes(EDUF):activation of the AMPK pathway[J]. Food Chem,2014,147:124-130.
[12] HE R,GIRGIH A T,ROZOY E,et al. Selective separation and concentration of antihypertensive peptides from rapeseed protein hydrolysate by electrodialysis with ultrafiltration membranes[J]. Food Chem,2016,197(Pt A):1 008-1 014.
[13]中华人民共和国卫生部.保健食品检验与评价技术规范(2003版)[S].北京:中国标准出版社,2003.
[14]魏稚萍,张才骏,杨荣珍,等.大通家牦牛及含野血牦牛血清亲血色蛋白多态性的比较研究[J].草食家畜,1996,1:22-23.
[15]张才骏,马凤莲,李黎,等.互助白牦牛血液蛋白质多态性的研究[J].青海畜牧兽医杂志,1996,26(3):9-11.
[16]魏稚萍.大通家牦牛及含野血牦牛血清运铁蛋白多态性的比较研究[J].甘肃畜牧兽医,1997,5:18-20.
[17]姚星辰,许博林,铁军,等.牦牛活性蛋白对小鼠抗缺氧抗疲劳能力及血糖的影响[J].时珍国医国药,2014,25(11):2 637-2 639.
[18]李冬利,于洪儒,杨菁,等.牛血活性肽对缺氧损伤心肌细胞的保护作用[J].中国新药杂志,2006,14:1 169-1 172.
[19]杜昕,李诚,肖岚,等.酶解牦牛血发酵液制备抗氧化肽工艺优化[J].核农学报,2017,31(2):325-333.
[20]杜昕,肖岚,李诚,等.枯草芽孢杆菌发酵牦牛血制备抗氧化肽工艺优化[J].食品与机械,2016,32(3):165-168; 205.
[21] TIAN F,LI B,JI B P,et al. Antioxidant and antimicrobial activities of consecutive extracts from Galla chinensis:The polarity affects the bioactivities[J]. Food Chemistry,2009,113(1):173-179.
[22] CHEN Naifu. Study on the extraction of flavonoid compound in pteridium aquilinum and its antioxidant property[J]. Food and Fermentation Industries,2003,29(11):63-67.
[23]刘昭明,黄翠姬,孟陆丽,等.核桃蛋白酶解物的抗氧化活性研究[J].食品与发酵工业,2009,35(1):58-61
[24]潘瑶,郑时莲,邹兴平,等.葡萄、芒果、草莓乙醇提取物抗氧化活性组分分析及其抗氧化相互作用[J].食品科学,2017,04:133-140.
[25]朱夕波.鲨鱼皮和猪皮胶原蛋白及其抗氧化活性酶解物的特性研究[D].上海:上海海洋大学,2008.
[26] YANG M,SHEN Q,LI L Q,et al. Phytochemical profiles,antioxidant activities of functional herb Abrus cantoniensis and Abrus mollis[J]. Food Chemistry,2015,177:304-312.
[27]张晖,唐文婷,王立,等.抗氧化肽的构效关系研究进展[J].食品与生物技术学报,2013,32(7):673-679.
[28]丁顺杰,罗金凤,丁晓雯,等.酶解缫丝蚕蛹蛋白抗氧化肽的分离与稳定性研究[J].食品科学,2015,36(3):35-40.
[29] CHI C F,CAO Z H,WANG B,et al. Antioxidant and functional properties of collagen hydrolysates from Spanish mackerel skin as influenced by average molecular weight[J]. Molecules,2014,19(8):11 211-11 230.
[30] SUETSUNA K,CHEN J R. Isolation and characterization of peptides with antioxidant activity derived from wheat gluten[J]. Food Science and Technology Research,2002,8(3):227-230.
[31]徐力,李相鲁,吴晓霞,等.一种新的玉米抗氧化肽的制备与结构表征[J].高等学校化学学报,2004,25(3):466-469.
[32] QIAN Z J,JUNG W K,KIM S K. Free radical scavenging activity of a novel antioxidative peptide purified from hydrolysate of bullfrog skin,Rana catesbeiana Shaw[J].Bioresource Technology,2008,99(6):1 690-1 698.
[33] SAMONINA G,ASHMARIN I,LYAPINA L. Glyproline peptide family:review on bioactivity and possible origins[J]. Pathophysiology,2002,8(4):229-234.
[34] BAILEY D M,BARTSCH P,KNAUTH M,et al. Emerging concepts in acute mountain sickness and high-altitude cerebral edema:from the molecular to the morphological[J]. Cell Mol Life Sci,2009,66:3 583-3 594.
[35] SCHREIBER M,TROJAN S. Protective effect of flavonoids and tocopherol in high altitude hypoxia in the rat:comparison with ascorbic acid[J]. Cesk Fysiol,1998,47:51-52.
[36] AL-HASHEM F H. Potential roles for vitamins E and C in combination in modulating exhaustive swimming and high altitude-associated lung injury in rats[J]. Saudi Med J,2012,33:367-374.
[37] FAN Pengcheng,MA Huiping,JING Linlin,et al. The antioxidative effect of a novel free radical scavenger 4'-hydroxyl-2-substituted phenylnitronyl nitroxide in acute highaltitude hypoxia mice. Biological&Pharmaceutical Bulletin,2013,36(6):917-924.
[38] MASTER S,GOTTSTEIN J,BLEI A T. Cerebral blood flow and the development of ammonia-induced brain edema in rats after portacaval anastomosis[J]. Hepatology,1999,30:876-880.