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1、第38卷第3期貴陽醫(yī)學(xué)院學(xué)報(bào)VO1．38No．32013年6月JOURNALOFGUIYANGMEDICALCOLLEGE2013．6貴k,l·I豆豉纖溶酶發(fā)酵液中活性酶的鹽析分離純化張敏，宋智魁，王鵬嬌，王清忱，彭安堂。，舒陽，高秀麗(1．貴陽醫(yī)學(xué)院藥學(xué)院，貴州貴陽550004；2．貴州省人民醫(yī)院，貴州貴陽550004；3．石家莊市中醫(yī)院，河北石家莊050051)【摘要J目的：對(duì)貴州豆豉發(fā)酵液中纖溶酶分離純化。方法：貴州特色藥食兩用的豆豉中提取得到的纖溶酶菌株(DCK—B)，用液體發(fā)酵法將其發(fā)酵，采
2、用硫酸銨鹽析法進(jìn)行粗提純，并對(duì)0～80％濃度的一級(jí)硫酸銨梯度鹽析與0～30％、30％～60％兩段硫酸銨梯度鹽析的結(jié)果進(jìn)行比較。結(jié)果：硫酸銨梯度鹽析在30％以前出現(xiàn)的沉淀為雜蛋白，且較復(fù)雜；30％梯度以后的目標(biāo)蛋白大量沉淀出來，到50％時(shí)活性蛋白幾乎沉淀完全；經(jīng)兩段鹽析，當(dāng)0—30％鹽析時(shí)有少量活性蛋白鹽析出來，絕大部分蛋白在30％～60％這個(gè)鹽析梯度沉降下來。結(jié)論：確定了貴州豆豉纖溶酶發(fā)酵液中活性酶的兩段硫酸銨鹽析分離純化方法。[關(guān)鍵詞]貴州豆豉；纖溶酶；硫酸銨鹽析；提純[中圖分類號(hào)]Q556．9[文
3、獻(xiàn)標(biāo)識(shí)碼]A[文章編號(hào)]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世界衛(wèi)生組織公布的數(shù)據(jù)顯示，心腦血管病例日益增多，已超過腫瘤和糖尿
