吉林大学学报(工学版) ›› 2012, Vol. 42 ›› Issue (增刊1): 466-469.
刘静波1, 王菲1, 张燕1, 王二雷1, 王作昭1, 姜轶群2
LIU Jing-bo1, WANG Fei1, ZHANG Yan1, WANG Er-lei1, WANG Zuo-zhao1, JIANG Yi-qun2
摘要： 为了制备安全性高、副作用小的凝血酶抑制肽,将蛋清Alcalase酶解产物经凝胶柱色谱Sephadex G-50凝胶柱色谱和半制备高效液相色谱纯化,旋转蒸发、冷冻干燥后经酶标仪法检测其抗凝血活性,筛选出高抗凝血活性的组分。结果表明:经Sephadex G-50纯化后的第4个组分即洗脱时间为35～40 min的组分抗凝血活性最好,抑制率达到84.74%。将该组分通过半制备高效液相色谱纯化后得到12个峰,峰Ⅶ 即洗脱时间为17.275～19.542 min的组分活性最高,抑制率达到71.27%。
中图分类号:
| [1] 新华社. 心脑血管病成为我国发病率死亡率最高疾病..http:[C]//www.ebiotrade.com/newsf/2009-3/200932393459.html.[2] Magalhaes A , Magalhaes H P, Richardson M,et al. Purification and properties of a coagulant thrombin-like enzyme from the venom of Bothrops leucurus[J]. Comparative Biochemistry and Physiology, 2007, 146(4):565-575.[3] Kong Yi, Huo Jian-li, Xu Wen, et al. A novel anti-platelet aggregation tripeptide from Agkistrodon acutus venom: Isolation and characterization[J]. Toxicon, 2009, 54(2):103-109.[4] Sanchez E E, Rodriguez-Acosta A, Palomar R, et al. Colombistatin: a disintegrin isolated from the venom of the South American snake (Bothrops colombiensis) that effectively inhibits platelet aggregation and SK-Mel-28 cell adhesion[J]. Archives of Toxicology, 2009, 83(3):271-279.[5] Barbouche R, Marrakchi N, Mansuelle P, et al. Novel anti-platelet aggregation polypeptides from Vipera lebetina venom: Isolation and characterization[J]. Febs Letters, 1996, 392(1):6-10.[6] Osipov A V, Filkin S Y, Makarova Y V, et al. A new type of thrombin inhibitor, noncytotoxic phospholipase A2, from the Naja haje cobra venom[J]. Toxicon, 2010, 55(2-3):186-194.[7] Giron M E, Estrella A, Sanchez E E, et al. Purification and characterization of a metalloproteinase, Porthidin-1, from the venom of Lansberg's hog-nosed pitvipers (Porthidium lansbergii hutmanni)[J]. Toxicon, 2011, 56(4):608-618.[8] Won-Kyo J, Se-Kwon K. Isolation and characterisation of an anticoagulant oligopeptide from blue mussel, Mytilus edulis[J]. Food Chemistry, 2009, 117(4):687-692.[9] Gan W Q, Deng L, Yang Chen, et al. An anticoagulant peptide from the human hookworm, Ancylostoma duodenale that inhibits coagulation factors Xa and XIa[J]. Febs Letters, 2009, 583(12):1976-1980.[10] Vija H, Samel M, Siigur E, et al. Purification, characterization, and cDNA cloning of acidic platelet aggregation inhibiting phospholipases A2 from the snake venom of Vipera lebetina (Levantine viper)[J]. Toxicon , 2009, 54(4):429-439.[11] Jo H Y, Jung W K, Kim S K, et al. Purification and characterization of a novel anticoagulant peptide from marine echiuroid worm, Urechis unicinctus[J]. Process Biochemistry, 2008, 43(2):179-184.[12] 田刚,陈代文,余冰,等. 酶解鸡蛋清小肽混合物对小鼠免疫功能的影响[J]. 中国畜牧杂志,2005,41(5):14-17. Tian Gang, Chen Dai-wen, Yu Bing, et al. Effects of mixtures of small peptides from enzymic hydrolysates of egg white on immune functions of mice[J]. Chinese Journal of Animal Science, 2005, 41(5):14-17.[13] Miguel Marta, Lopez-Fandino Rosina, Ramos Mercedes, et al. Long-term intake of egg white hydrolysate attenuates the development of hypertension in spontaneously hypertensive rats[J]. Life Science, 2006, 78(25):2960-2965.[14] Yang W G, Wang Z, Xu S Y. A new method for determination of antithrombotic activity of egg white protein hydrolysate by microplate reader[J]. Chinese Chemical Letters, 2007, 18(4):449-451. |
| [1] | 刘静波, 王菲, 王翠娜, 刘军, 王作昭, 王二雷, 张燕, 林松毅. Alcalase酶解蛋清粉制备抗凝血肽的工艺优化[J]. 吉林大学学报(工学版), 2012, 42(01): 250-255. |
|
||