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1、314食品與藥品FoodandDrug2013年第15卷第5期產(chǎn)劍麻皂苷水解酶菌株的篩選與鑒定鄧天發(fā),王京博,謝慶武(廣西大學(xué)生命科學(xué)與技術(shù)學(xué)院,廣西南寧530004)摘要:目的篩選能酶解劍麻皂苷并釋放游離劍麻皂素的菌株。方法用薄層色譜法(TLC)初篩能酶解劍麻皂苷游離出劍麻皂素的菌株,再用酶活性比色測(cè)定和高效液相色譜法(HPLC)復(fù)篩目的菌株。通過形態(tài)學(xué)觀察和ITS序列測(cè)定,初步鑒定菌種。結(jié)果篩選~lJ31株目的菌株,其中絲狀真菌13株,細(xì)菌18株。絲狀真菌的酶活性高于細(xì)菌。鑒定其中酶活性較高的4株真菌:菌種T1為黑曲霉,T2、T
2、3為鏈孢霉屬,T4為青霉屬。結(jié)論TLC可快速篩選酶解劍麻皂苷并釋放游離劍麻皂素的菌種。黑曲霉、鏈孢霉和青霉均有較強(qiáng)的酶解劍麻皂苷并釋放游離劍麻皂素的能力。關(guān)鍵詞:劍麻皂素;皂苷水解酶;菌種;篩選中圖分類號(hào):Q93.331文獻(xiàn)標(biāo)識(shí)碼:A文章編號(hào):1672—979X(2013)05—0314-03ScreeningandIdentificationofSisalanaSaponinHydrolyticEnzymeProducingStrainsDENGTian—fa,WANGJing—bo,XIEQing—WH(CollegeofLif
3、eScienceandTechnology,GuangxiUniversity,Nanning530004,China)Abstract:ObjectiveToscreenthestrainswhichcanhydrolyzetheagavesisalanasaponinandreleasethetigogenin.MethodsUsingtheTLCassay,thestrainsthatcanhydrolyzetheagavesisalanasaponinandreleasethetigogeninwerefirstscreen
4、edout,thenthedestinationstrainswerescreenedoutagain,usingtheenzymaticcolorimetricdeterminationandHPLCdetermination.BymorphologicalobservationandITSsequencing,thestrainswerepreliminaryidentified.Results31destinationstrainswerescreenedout.a(chǎn)mongwhich13strainswerefilamento
5、usfungiand18werebacteria.Theactivityofthehydrolyticenzymeofthefungiwashigherthanthatthebacteria.The4fungistrainsT1,T2,T3andT4withhigherenzymaticactivitywereidentifiedasAspergillusniger,Neurosporasitophila,andPenicillium,respectively.ConclusionUsingtheTLCassay,thescreen
6、ingofthestrainsthatcanhydrolyzetheagavesisalanasaponinandreleasethetigogeninwasaccelerated.Aspergillusniger,Neurosporasitophila,andPenicilliumallhavethecapacitytohydrolyzetheagavesisalanasaponinandreleasethetigogenin.KeyWords:tigogenin;saponinhydrolyticenzyme;strain;sc
7、reening劍麻皂素(tigogenin)具有抗炎、解熱、鎮(zhèn)痛、增強(qiáng)學(xué)農(nóng)場(chǎng)等地采集。免疫、降低血糖、抗腫瘤、抗菌、降血脂、降血壓等作1.2培養(yǎng)基用,應(yīng)用廣泛,可用于合成120多種激素類藥物,還可應(yīng)用平板篩選培養(yǎng)基:劍麻渣粉末30g,NaNO,2g,于食品、化妝品及畜牧業(yè)等行業(yè)。]。劍麻皂素在劍麻中以K2HPO41g,KC10.5g,MgSO4’7H2O0.5g,F(xiàn)eSO4‘7H2O甾體皂苷配基的形式存在,在其C3位~NC26位等通過糖苷0.01g,瓊脂12g,水1000mL,pH自然。鍵與糖鏈(由葡萄糖、木糖、鼠李糖、半乳糖、阿拉
8、伯糖產(chǎn)酶培養(yǎng)基:劍麻渣粉末1g,葡萄糖O.25g,蛋白等組成)相連]。制備劍麻皂素,必須先水解與其直接相胨0.125g,K2HPO4O.025g,KC1O.0125g,MgSO4’7H2O連的糖苷鍵(以下簡(jiǎn)稱皂苷鍵)。目前,國內(nèi)提取劍