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	克鲁维酵母Y-85菊粉酶的纯化和性质; PURIFICATION AND PROPERTIES of INULINASE FROM KLUYVEROMYCES SP. Y-85
	魏文铃 ; 余娴文 ; 戴亚 ; 郑晶 ; 谢忠
	1997
关键词	克鲁维酵母 菊粉酶 纯化和性质 Kluyveromyces Inulinase PuriFication and properties
英文摘要	克鲁维酵母(kluyVErOMyCESSP.)y-85产生的胞内菊粉酶(EndOCEllulArInulInASE)和胞外菊粉酶(EXOCEllulArInulInASE)粗酶液分别经PEg6000-磷酸盐缓冲液双水相抽提得部分纯化酶液。前者进一步用硫酸铵分级沉淀、PrOTEIn-PAkdEAE离子交换、PrOTEIn-PAk200SW凝胶过滤后得到两个菊粉酶组分EⅠ和EⅡ;后者采用dEAE-SEPHACEl离子交换、SEPHAdEXg150凝胶过滤后得到菊粉酶EEXO。经WATErS650E蛋白纯化系统鉴定,三者均呈单一的对称峰;EⅠ和EⅡ达聚丙烯酰胺盘状凝胶电泳纯。EⅠ、EⅡ和EEXO的分子量分别为42kd、65kd和57kd;三者均为糖蛋白,多糖含量分别为30%、35%和25%;I/S(InulInASEACTIVITy/SuCrASEACTIVITy)比值分别为0.086、0.078和0.072;三者均属外切菊粉酶。EⅠ、EⅡ和EEXO酶反应最适PH分别为4.6、4.5和4.6,最适温度分别为52℃、52℃和55℃;Ag--+、Hg--(2+)和PCMb对酶活性有强烈的抑制作用;三者水解菊芋粉糖液的产物均为果糖(86.5%)和葡萄糖(13.5%)。; The crude endocellular inulinase From Kluyveromyces sp.Y-85 was puriFied to two components, designated as EⅠ and EⅡ, using PEG6000-phosphate buFFer extraction, (NH4)2SO4 Fractionation, DEAE chromatography and gel Filtration (Protein-PAK); The crude exocellular inulinase From this strain was puriFied to Eexo by means of PEG6000-phosphate buFFer extraction, double DEAE-Sephace chromatography, Sephadex G-150 gel Filtration.EⅠ, EⅡ and Eexo were demonstrated to be homogeneous by Waters 650E protein puriFication system.Thier molecular weights are 42kD, 65kD and 57kD, respectively.All the inulinases were glycoproteins containing a saccharide (From 25% to 35%) and belonged to the endo-inulinase.In addition, E Ⅰ, EⅡ, Eexo were optimally reactive at pH4.6,4.5,4.6 and at 52℃ , 52℃ , 55℃,respectively.Ag+, Hg2 + and PCMB inhibited these enzymes' activity strongly.The products of raw inulin extracted From Helianthus tuberosus hydrolyzed by these three enzymes were Fructose (86.5%) and glycose (13.5%); 国家“八·五”攻关项目资助课题
语种	zh_CN
内容类型	期刊论文
源URL	[http://dspace.xmu.edu.cn/handle/2288/119388]
专题	生命科学－已发表论文
推荐引用方式 GB/T 7714	魏文铃,余娴文,戴亚,等. 克鲁维酵母Y-85菊粉酶的纯化和性质, PURIFICATION AND PROPERTIES of INULINASE FROM KLUYVEROMYCES SP. Y-85[J],1997.
APA	魏文铃,余娴文,戴亚,郑晶,&谢忠.(1997).克鲁维酵母Y-85菊粉酶的纯化和性质..
MLA	魏文铃,et al."克鲁维酵母Y-85菊粉酶的纯化和性质".(1997).

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