小体系高通量比色测定糖化酶活力新方法.pdfVIP

2012 Vol.31 No.7 .14Qo Serial No.244 China Brewing Innovation and Knowledge Transfer 小体系高通量比色测定糖化酶活力新方法 刘 洋1 ，唐芳荣1 ，*目宏宇1 ，朱洪亮2 ，谢秋宏1* (1.吉林大学生命科学学院，吉林长春 130012; 2. 淄博国澳生物酶技术有限公司，山东淄博 255ω0) 摘 要2 本研究建立了一种小体系结合96孔板比色测定糖化酶活力的新方法。经实验证明，在96孔板内，波长5咽nm处，葡萄糖浓度 与光密度(OD)值有很好的线性关系，其线性相关系数r值达到0.9996: 通过加样回收实验发现，其回收率为98.5~♀-103.2%，相对标准 差为1.8%; 通过精密度分析实验发现该方法测量样品的精密度优于国标法及普通DNS法，其测定的结果与现行国家标准测定糖化酶 方法所得到的结果差异率小于0.5%。本研究建立的方法不仅准确、可靠，而且实现了小体系、高通量快速比色测定，为糖化酶工业检 测及糖化酶生产菌株的高通量筛选奠定了重要方法基础。 关键词z 糖化酶:小体系:高通量:活力测定:比色法 中图分类号:Q5562 文献标识码:A 文章编号，0254-5071(2012)07-014ι03 Anewme也odofmi臼0-町蜘mandhigh..:也roughput determination of glucoamylase activity by Colorim甜y l LIUY皿gl， TANG Fangrong ,XIANG Hongyul , ZHU Hongliang2, XIE Qiuhon.俨 (J.Col/ege ofLife SIαence， Jilin Universii伊; Changchun 13ω12， C抽8; 2. Zibo Guoao Biological Enzyme Technology Limi衍d Company, Zi加2551α刃~ China) Abs1ract:In也is 阳dy， we repo由d a new assay method 伽.t a micro-system detennined the activity of g1ucoamylase by dinitrosalicylic acid (DNS) reaction. In 96-well plates, glucose concentration and optical density (OD) values showed linear relationship at 540nm, and the linear correlation coe伍­ cient (r) values was 0.9996. The 臼mplere∞，very experiment described that 也erecov笛y range and the rela咀明白且也rd deviation 但SD)w部98.50/ 103.2% and 1.8%, respectively. The precision analysis to confum the method is accurate and reliable. To compared with national sta且也rd method, the di能rences in rates ofglucoamylase activityωsay was less than 0.5%.白le method is rapíd, accurate and high也roughput on determination ofg1ucoamy- lωe activity, and will become an important method on quality control of commodity glucoamylase and screening of glucoamylase producing s町ain. K町W惯也: glucoa