作　　者： ; ; ; ; ;

机构地区： 广西大学生命科学与技术学院

出　　处： 《生物加工过程》 2010年第4期62-66,共5页

摘　　要： 分离得到1株产生淀粉酶的菌株,通过扩增和测定16S rDNA序列并进行比对,发现是Paenibacillus属的细菌。液体摇瓶发酵结束后,其产生的生淀粉酶比酶活达108.5U/mL。通过饱和(NH4)2SO4沉淀、Sephacryl S-300层析的方法对其所产的生淀粉酶进行分离纯化,得到纯化的酶蛋白比酶活为5112.04U/mg,纯化倍数为14.1,相对分子质量约为1.0×105。该酶以木薯生淀粉为底物时,最适pH5.6,最适温度50℃。金属离子Ca2+、Mg2+对该酶具有激活作用,Zn2+、Cu2+、Fe2+、Ni2+、Mn2+、Co2+和EDTA2-对该酶均具有抑制作用。 An isolated raw starch-digesting amylase producing strain was cultured in liquid media and identified as Paenibacillus sp.by amplification,sequencing and alignment analysis of 16S ribosomal DNA sequence.In shaking flask culture,the yield of the amylase from Paenibacillus sp.reached 108.5 U/mL.The amylase was purified from culture supernatant of Paenibacillus sp.by ammonium sulphate precipitation and Sephacryl S-300 chromatography.14.1-Fold increase in purity with specific activity 5 112.04 U/mg proteins was obtained.The apparent molecular weight of the purified enzyme was 1.0×105 by SDS-PAGE.Enzymatic characterization revealed that optimum activity was pH 5.6 and temperature of 50 ℃.The enzyme was activated by Ca2＋ and Mg2＋,while inhibited by Zn2＋,Cu2＋,Fe2＋,Ni2＋,Mn2＋,Co2＋,and EDTA2-,respectively.

关 键 词： 生淀粉酶 分离纯化

领　　域： [生物学]