坎皮纳斯类芽孢杆菌木聚糖酶的纯化以及酶学性质  

Purification and enzymatic properties of xylanase from Paenibacillus campinasensis

作　　者：陈磊[1] 赵祥颖[2] 刘建军[1,2] 

机构地区：[1]齐鲁工业大学生物工程学院,山东济南250013 [2]山东省食品发酵工程重点实验室,山东济南250013

出　　处：《中国酿造》2016年第7期64-68,共5页China Brewing

基　　金：山东省自主创新专项(201422cx02602)

摘　　要：采用硫酸铵盐析、Sephadex G-50凝胶过滤、DEAE Sepharose Fast Flow离子交换层析对坎皮纳斯类芽孢杆菌(Paenibacillus campinasensis)xy-7发酵液进行分离纯化,获得纯化的木聚糖酶,纯化倍数为26.36,酶活回收率为5.13%。十二烷基硫酸钠聚丙烯酰胺凝胶电泳(SDS-PAGE)结果为单一条带,分子质量为24.5 ku。酶学性质研究结果表明,该酶的最适反应温度为60℃,最适p H值为8.0。K+、Fe2+对酶有激活作用,Zn2+、Cu2+对酶有强烈的抑制作用。以榉木木聚糖为底物时,米氏常数Km=4.733 mg/m L,最大酶反应速率Vm=315.85μmol/(min·mg)。The fermentation liquid of Paenibacillus campinasensis xy-7 was separated and purified by ammonium sulfate salting-out, Sephadex G-50 gel filtration and DEAE Sepharose Fast Flow ion-exchange column chromatography. The purified xylanase was obtained, the purification multi- ple was 26.36 and recovery rate of the enzyme was 5.13%. The electrophoresis result of SDS-PAGE was single band and mass of molecule was 24.5 ku. The results of enzymatic property showed that the optimal reaction temperature of xylanase was 60℃ and the optimal pH was 8.0. The enzyme could be activated by Fe2＋ and K＋, but inhibited by Zn2＋ and Cu2＋. With beech xylan as substrate, the Km was 4.733 rag/m1 and the Vm was 315.85 μmol/（min·mg）.

关 键 词：坎皮纳斯类芽孢杆菌 木聚糖酶 分离纯化 酶学性质 

分 类 号：Q814.1[生物学—生物工程]