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1. 发明授权

US5449615A KDN-cleaving sialidase isolated from hepatopancreas of mollusks 失效
标题翻译：从软体动物肝胰腺分离的KDN切割唾液酸酶
公开(公告)号：US5449615A
公开(公告)日：1995-09-12
申请号：US311957
申请日：1994-09-26
申请人： Yuh-Teh Li , Su-Chen Li
发明人： Yuh-Teh Li , Su-Chen Li
IPC分类号： C12N9/24 , C12N9/36
CPC分类号： C12Y302/01129 , C12N9/2402 , C12Y302/01018
摘要： A sialidase capable of cleaving a glycoconjugate to yield 3-deoxy-D-glycero-D-galacto-2-nonulosonic acid (KDN) is isolated from hepatopancreas of mollusks. Preferably, the sialidase is isolated by processing crude oyster hepatopancreas to form a crude extract of the sialidase enzyme. The crude enzyme is further concentrated and refined utilizing sequential column chromatography fractogel EMD DEAE-650 (M), sephacryl S-200, and fractogel EMD SP 650 (M) chromatography, respectively. Using this procedure, the sialidase is purified over 155 fold with a 17% recovery. Ferric ion exerts a three fold stimulation in the activity of the sialidase, at a concentration of 3 mM. The sialidase cleaves 4-methylumbelliferyl-KDN, KDN.alpha.2.fwdarw.3Gal.beta.1.fwdarw.4GlcCer, KDN.alpha.2.fwdarw.6Gal.beta.1.fwdarw.4GlcCer, or KDN.alpha.2.fwdarw.6GalNAc-o1 to yield KDN.
摘要翻译：能够切割糖缀合物以产生3-脱氧-D-甘油-D-半乳-2-异壬酸（KDN）的唾液酸酶从软体动物的肝胰腺中分离。优选地，通过加工粗牡蛎肝胰腺来分离唾液酸酶以形成唾液酸酶的粗提取物。使用顺序柱色谱分离凝胶EMD DEAE-650（M），sephacryl S-200和分子凝胶EMD SP 650（M）色谱法分离粗酶。使用该方法，将唾液酸酶以17％的回收率纯化155倍。铁离子以3mM的浓度对唾液酸酶的活性施加三倍的刺激。唾液酸酶切割4-甲基伞形酮基KDN，KDNα2->3Galβ1-> 4GlcCer，KDNα2->6Galβ1-> 4GlcCer或KDNα2-> 6GalNAc-o1以产生KDN。

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