贵州豆豉纤溶酶发酵液中活性酶的盐析分离纯化-论文.pdf

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1、第38卷第3期贵阳医学院学报VO1．38No．32013年6月JOURNALOFGUIYANGMEDICALCOLLEGE2013．6贵k,l·I豆豉纤溶酶发酵液中活性酶的盐析分离纯化张敏，宋智魁，王鹏娇，王清忱，彭安堂。，舒阳，高秀丽(1．贵阳医学院药学院，贵州贵阳550004；2．贵州省人民医院，贵州贵阳550004；3．石家庄市中医院，河北石家庄050051)【摘要J目的：对贵州豆豉发酵液中纤溶酶分离纯化。方法：贵州特色药食两用的豆豉中提取得到的纤溶酶菌株(DCK—B)，用液体发酵法将其发酵，采

2、用硫酸铵盐析法进行粗提纯，并对0～80％浓度的一级硫酸铵梯度盐析与0～30％、30％～60％两段硫酸铵梯度盐析的结果进行比较。结果：硫酸铵梯度盐析在30％以前出现的沉淀为杂蛋白，且较复杂；30％梯度以后的目标蛋白大量沉淀出来，到50％时活性蛋白几乎沉淀完全；经两段盐析，当0—30％盐析时有少量活性蛋白盐析出来，绝大部分蛋白在30％～60％这个盐析梯度沉降下来。结论：确定了贵州豆豉纤溶酶发酵液中活性酶的两段硫酸铵盐析分离纯化方法。[关键词]贵州豆豉；纤溶酶；硫酸铵盐析；提纯[中图分类号]Q556．9[文

3、献标识码]A[文章编号]1000．2707(2013)03-0235-04SeparationandPurificationofActiveEnzymefromFibrinolyticEnzymeFermentationBrothofGuizhouLobsterSaucewithSaltingFractionationMethodZHANGMin，SONGZhikui，WANGPengjiao，WANGQingchen。，PENGAntang，SHUrang，GAOXiuli(．SchoolofPha

4、rmacology，GuiyangMedicalCollege，Guiyang550004，Guizhou，China；2．PeopleSHospitalofGu&houProvince，Guiyang550002，Gu~hou，China；3．TraditionalChineseMedicineHospitalofShijiazhuangCity，Shijiazhuang050051，Hebei，China)[Abstract]objective：Toculturefibrinolyticenzym

5、estrains(DCK—B)extractedfromGuizhoucharacteristiclobstersaucewithliquidfermentmethod，andtodevelopamethodforprimarypufifica—tionofactivefibrinolyticenzymefromthefermentationproducts．Methods：Ammoniumsulphate(AS)saltingoutmethodwasusedforprimarypurificatio

6、n，andtheeffectsofonestepsaltingoutmethodwith0％一80％ASandtwostepsaltingoutmethodwithASof0％～30％and30％一60％werecom-pared．Results：TheprecipitatewasimpureproteinwhenASconcentrationwaslowerthan30％．AlotoftargetproteinprecipitatedwhenASconcentrationwashigherthan3

7、0％．Whentheconcentrationwasupto50％，almostalltheactiveproteinprecipitated．ItindicatedthatafewactiveproteinwassaltedoutwhenASconcentrationwasbetween0—-30％andmostofactiveproteinsaltedoutwhenAScon—eentrationwasbetween30％～60％．Conclusions：ThetwostepASsaltingou

8、tmethodforseparationandpurificationofactiveenzymefromfibrinolyticenzymefermentationbrothofGuizhoulobstersauceisestablished．[Keywords]GuizhouDouchi；plasmin；ammoniumsulfateprecipitation；purification世界卫生组织公布的数据显示，心脑血管病例日益增多，已超过肿瘤和糖尿