基于pHBH法的岩藻聚糖硫酸酯酶酶活测定方法  

Fucoidanase Activity Determination Method on Basis of pHBH Method

作　　者：张翠玉[1] 薛长湖[1] 于龙[1] 王彦超[1] 续晓琪[1] 常耀光[1] 

机构地区：[1]中国海洋大学食品科学与工程学院,山东青岛266003

出　　处：《中国食品学报》2013年第7期200-206,共7页Journal of Chinese Institute Of Food Science and Technology

基　　金：国家自然科学基金项目(31101302);教育部博士点基金项目(20110132120015)

摘　　要：采用对羟基苯甲酰肼(pHBH)对岩藻糖进行显色,建立测定岩藻糖还原端含量的pHBH法。该方法操作简便,灵敏度高,检测限为0.26μg/mL,定量限0.85μg/mL。在此基础上,以Flavobacteriaceae sp.CZ1127产生的岩藻聚糖硫酸酯酶为试验材料,对岩藻聚糖硫酸酯酶活测定方法中的几个关键影响因素进行研究,建立一套快速、可靠的岩藻聚糖硫酸酯酶酶活测定方法。采用该方法,酶活检测限达到0.048 mU。In this study, a new method using 4-Hydroxyben zhydrazide for the content determination of fucose reducing end was establided at frist. The detection limit of this method was 0.26 μg/mL and the quantification limit was 0.85 μg/mL. It possessed several advantages, including simple-operating, high precision and sensitivity. On base of this, we study some key factors for the fueoidanase activity detection. Then we built a fast and reliable fueoidanase activity detection method as follows： a mixture of 375μL fucoidan solution with the concentration of 0.055% and 375 μL enzyme solution was frist reacted at 28℃ for 30minutes. After that 250 μL of pHBH was added in, the mixture was then reacted at 100 ℃ for 5 minutes. After cooling off, the absorbance was detemined at 415 nm. Using this method, the fueoidanase activity detection limit was as low as 0.048 mU.

关 键 词：羟基苯甲酰肼 岩藻糖 岩藻聚糖硫酸酯酶 糖苷酶 酶活测定 

分 类 号：TS207.3[轻工技术与工程—食品科学]