聚丙烯酰胺凝胶固定化酶法制备麦芽四糖的研究  

Study on Preparation of Maltotetraose by Using Polyacrylamide Gel Immobilized Enzyme Method

作　　者：唐玉[1] 李东霄[1] 梁新红[1] 李元召[1] 孙俊良[1] 

机构地区：[1]河南科技学院食品学院,河南新乡453003

出　　处：《江西农业学报》2013年第7期88-90,共3页Acta Agriculturae Jiangxi

基　　金：国家自然科学基金项目(C200101)

摘　　要：采用不连续聚丙烯酰胺凝胶电泳(PAEG)对麦芽四糖淀粉酶粗酶液进行分离纯化,经质谱分析确定了麦芽四糖酶条带,然后切胶回收获得高纯度的麦芽四糖酶。将获得的含有麦芽四糖酶的凝胶制成颗粒通过甲醛链接固定,获得固定化麦芽四糖酶。经薄层色谱和液相色谱分析发现:使用固定化酶生产的麦芽四糖纯度高于粗酶液,尤其是单糖杂质含量显著减少。此外,固定化麦芽四糖酶的比活力升高。A discontinuous polyacrylamide gel electrophoresis (PAEG) was performed to isolate and purify the maltotetraose -producing amylase from their crude preparations .The bands containing amylase in the gel were detected by mass spectral analysis , and the bands containing high -purity maltotetraose -producing amylase were cut down from the gel .The gel containing maltote-traose-producing amylase was squeezed into small pellets , and then immersed in 10% formaldehyde for the linkage between gel and enzyme as well as the immobilization of the amylase , finally the immobilized maltotetraose -producing amylase was obtained .By the analysis of thin layer chromatography and high performance liquid chromatography , the maltotetraose produced by immobilized enzyme had higher purity than that did by the crude enzyme solution , especially it had a significant decrease in monosaccharide con-tent.In addition, our results also indicated that there was an increase in the specific activity of the immobilized maltotetraose -pro-ducing amylase.

关 键 词：麦芽四糖酶 固定化酶 聚丙烯酰胺凝胶电泳 麦芽四糖 

分 类 号：Q814.2[生物学—生物工程]