混合非淀粉多糖水解酶酶活测定方法的改进.pdfVIP

第 12 期 　　　　　　　2001 年 　　　　　 　　　　粮食与饲料工业 　　 　　　　　　　　　　　　　　　　　　　 45 检测分析 混合非淀粉多糖水解酶酶活测定方法的改进 史 　锋 ,王 　璋 (江南大学食品学院 , 江苏 无锡 　214036) β 摘 　要 :用分光光度法分别准确地测定了混合非淀粉多糖水解酶中木聚糖酶 、果胶酶 、- 葡聚糖酶和纤维素酶的酶 活 。但将酶添加到物料中后 ,酶活测定变得非常困难 ,于是对酶活测定方法进行了改进 。利用凝胶过滤色谱法对酶 液进行预处理 ,排除了体系中还原糖等小分子量杂质的干扰 ;采用琼脂放射扩散法测定纤维素酶或 β- 葡聚糖酶酶 活 ,提高了测定的特异性和灵敏度 ;利用专一性的酸性醋酸铜法测定果胶酶酶活 ,提高了特异性 ;采用硫酸铵沉淀法 处理木聚糖酶 ,可排除别的酶对木聚糖酶酶活测定的干扰 。 关键词 :非淀粉多糖水解酶 ;酶活测定 + ( ) 中图分类号 :TS 201. 2 5 ; TS 207 . 4 　　文献标识码 :A 　　文章编号 :1003 - 6202 2001 12 - 0045 - 03 Improvement of the Activity Determination of Complex Nonstarch Polysaccharides Hydrolases β ABSTRACT 　The activities of the nonstarch polysaccharide hydrolases , including xylanase , pectase , - glucanase and cellulase , in a premixed enzyme preparation were determined accurately by spectrophtometry. But when the premixed preparation was added into edibles , the enzyme activities could not be determined individually with accuracy. Some improvements of the determining meth ods were made as follows. Firstly , the enzyme solution was pretreated by gel filtration in order to remove fractions with low molecular β weight , such as reducing sugar . Secondly , through the agar radial diffusion , the sensitivity and specificity of - glucanase and cel lulase activity determination were improved greatly. In addition , the determining method of pectase activity was changed to more sensitive and