大蒜蒜酶提取纯化及临床应用研究

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1、人蒜蒜酶提取纯化及临床应用研究作者：曹红，陈坚单位：1•新疆医药大学药学院，新疆乌鲁木齐830054；2.石河子大学化学化工学院，新疆石河子832003【摘要】目的：研究从鲜蒜中分离纯化蒜臨的方法和临床常用剂対酶活性的影响，为今后工艺的进一步放大、优化和合理临床应用提供必要理论参数。方法：以天然蒜氨酸作底物，采用丙酮酸法测定酶浓度；以牛血清H蛋口为标准，采用考马斯亮兰G250法测定蛋口质浓度。结杲：从鲜蒜中提取纯化蒜酶，其粗酶浸提液的最佳组成为Na2HPO4KH2PO4缓冲溶液；硫酸较沉淀酶蛋白的授适饱和度范用

2、为0%〜35%；选用超滤技术脱盐；蒜酶故佳调节等电点为4.9。在临床常用溶剂中，含氯化钠的注射液(不含葡萄糖)对蒜酶活力有显著的激活作用，而含葡萄糖的注射液则较强地抑制了蒜酶活力；75%和95%乙醇具有强烈的杀酶作用。结论：本研究为今后工艺的进一步放大与优化和蒜酶合理临床应用提供了指导。【关键词】人蒜；蒜酶；提取纯化；活力；临床常川溶剂StudyofextractionandpurificationofalliinasefromgarlicanditsclinicalapplicationCAOHongl,2,

3、CHENJiaiil(l.DepaitmentofPharmacyofXinjiangMedicalUniversity,Urumqi830054,China；Z.ChemistryandChemicalengineeringCollegeofShiheziUniversity,Shihezi,832003,China)AbstractObjective：Thestudyofextractionandpurificationofalliinasefromgarlicandtheeffectofclinicals

4、olventcommonlyusedonenzymeactivitywereaccomplishedinthispape匚Itprovidesessentialparameterforfurtherpurificationprocedureandclinicalapplication.Methods：ProteinconcentrationsofallinasepreparationsweredetenninedaccordingtothemethodsofBradfordusintBSAasstandards

5、.EnzymeactivitywasassayedfromtheamountofenzymaticallyformedpyaivateusingnaturalL(+)alliinassubstrate.Results：Theprocessingconditionsfortheextractionandpurificationofalliinasefromgarlicwereoptimized.ltisinNa/Kphosphatebufferthatpeeledgarliccloveswerehomogeniz

6、edinablende匚Precipitationwithsolidammoniumsulfatetogive35%saturationwasselectedtoisolatealliinase.ltisbyultrafiltrationthatsupernatantsolutioncontainingalliinaseisdesalted.Theisoisnicpointofalliinasewasfoundtobe4.9.WheniiijectionswithNaClandwithoutglucoswasu

7、sedassolvent,therewasamarkedstimulationofenzymeactivity,butinjectionswithglucosgaveastronginhibition.Thelyasesolvedin75%or95%alcoholcompletelylostitsactivityrapidly.Conclusion：Thepresentpapergavedirectionstofurtherlyamplifyandoptimizepurificationprocedureofa

8、lliinaseanditsclinicalapplication.Keywordscarlic；alliinase；extractionandpurification；activity；clinicaldissolvantcommonlyused百合科葱属植物大^((AlliumsativumLin),是世界不同地区、不同民族公认的药食两用珍品，且历史悠久。在我国，一些著名的古医学文