响应面法优化鹿血血红蛋白酶解工艺的研究.docVIP

响应面法优化鹿血血红蛋白酶解工艺的研究 徐 馨1,2*韩欢胜2,3章许冉 （1. 黑龙江省兽医科学研究所， 黑龙江齐齐哈尔161005； 2. 东北农业大学哈尔滨150030； 3. 黑龙江省农垦科学院哈尔滨特产研究所， 黑龙江哈尔滨150038） 关键词： 摘要：鹿血血红蛋白酶解工艺影响蛋白酶解效果，包括酶解温度、酶解时间、酶添加量、pH值，二次回归正交旋转组合设计鹿血血红蛋白酶解的最佳工艺为酶解温度56 ℃、酶解时间3.78 h、酶添加量4.2 %、pH值=9.56。在此条件下，自由基清除率为（81.95 ± 1.63）%，酶解效果最优。响应面法用于鹿血血红蛋白酶解工艺的优化可行可靠，酶解条件适宜于本物系。Optimization of Enzymolysis Conditions of Hemoglobin from Deer Blood by Reverse Evaporation Methodology XU Xin1,2, HAN HuanSheng2,3, ZHANG Xuran3* （1. Heilongjiang Institute of Veterinary Science, Qiqihar Heilongjiang 161005, China；2. Northeast Agricultural University, Harbin Heilongjiang 150001, China；3. Heilongjiang Academy of Land Reclamation Sciences, Harbin Heilongjiang 150038, China） Keywords：deer blood；hemoglobin；polypeptide；enzymolysis；response surface methodology；clearance Abstract：grope the enzymolysis conditions of hemoglobin from deer blood, the response surface methodology was used to optimize their enzymolysis technology. On the basis of single factor tests, enzyme dosage, reaction time, reaction temperature and pH affecting DPPH clearance were firstly evaluated ,and then the enzymolysis conditions were optimized by response surface methodology.The optimal hydrolysis conditions were determined as hydrolyzing temperature was 56 ℃, reaction time was 3.78 h, enzyme dosage were 4.2 %, hydrolysis pH was 9.56. Under these conditions, the predictive value of DPPH clearance was up to （81.95 ± 1.63）%. It shows that the response surface analytical method is reliable and can be used in the process of enzymolysis optimization. 基金项目哈尔滨市青年后备人才项目（2015RQQYJ033）。 作者简介徐馨（1983-），助理研究员，博士，新药研发与临床诊疗。25xx99@163.com 通讯作者章许冉（197-），，，。zhangxuran2012@ 鹿血为鹿科动物梅花鹿(Cervus nippon Temminck.)或马鹿(Cervus elapH值us Linnaeus.)的膛血或茸血[1-2]。现代药理研究表明，鹿血具有抗衰老、增强和调节免疫、补气补血、补肾益精增强性功能、抗缺氧、抗疲劳、抗辐射、促进代谢、促进创伤愈合、对中枢神经抑制和降血压等多方面药理作用[3-5]。起到上述功能的主要活性物质为蛋白质和肽类化合物（又称生物活性肽，功能肽）。生物活性肽在微量或低浓度的情况下即可发挥生物调节作用，对维持人体正常的生理活动具有重要意义[6]。 为了实现对鹿血资源功效成分的高效富集，使其有益生物活性成分得以充分利用，由此获取高提取率、高纯度的生物活性肽，在精细分离纯化出鹿血中的生物活性肽成