SENSITIVE ENZYMATIC ASSAY FOR GLUCOSE DETERMINATION IN NATURAL-WATERS
SENSITIVE ENZYMATIC ASSAY FOR GLUCOSE DETERMINATION IN NATURAL-WATERS
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DOI：
10.1128/aem.33.6.1237-1243.1977
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发表时间：
1977-01-01
影响因子：
4.4
通讯作者：
PHELPS, G
中科院分区：
文献类型：
--
作者：
CAVARI, BZ;PHELPS, G
本文介绍了一种新的酶法测定葡萄糖的方法。它允许测量低至10-7 M的葡萄糖浓度，以了解微生物对营养物质的吸收。这种灵敏度使得该方法特别适合于估计天然水体中的葡萄糖，通常无需事先浓缩或提取。该方法基于葡萄糖和ATP之间的反应，由己糖激酶催化形成G-6-P。在该反应中消耗的ATP的量与样品中存在的葡萄糖的量成正比，通过胰蛋白酶-荧光素酶测定来测量。用该方法测定葡萄糖的最佳条件定义如下：在30 ℃下孵育20分钟。C、10-3 M的Mg浓度和7.5-10.5范围内的pH。还研究了该测定法对不同碳水化合物的专属性。添加到Kinneret湖[以色列]水中的已知量葡萄糖的回收率在80- 114%的范围内。该方法的应用证明在8个月的配置文件中的葡萄糖含量在Kinneret湖。
A new enzymatic method for glucose determination is described. It allows measurement of glucose concentration as low as 10-7 M in order to understand the uptake of nutrients by microorganisms. Such sensitivity makes this method particularly appropriate for estimation of glucose in natural-water bodies, generally without prior concentration or extraction. The method is based on the reaction between glucose and ATP, catalyzed by hexokinase to form G-6-P. The amount of ATP consumed in this reaction, which is directly proportional to the amount of glucose present in the sample, is measured by the luciferin-luciferase assay. The optimal conditions for glucose determination by this method were defined as follows: 20 min of incubation at 30.degree. C, Mg concentration of 10-3 M, and pH in the range of 7.5-10.5. The specificity of the assay to different carbohydrates was also studied. Recovery of known amounts of glucose added to Lake Kinneret [Israel] water was in the range of 80-114%. Application of this method is demonstrated in 8 monthly profiles of the glucose content in Lake Kinneret.