[使用哈维贝内克氏菌发光细菌的固定化提取物测定皮摩尔量烟酰胺腺嘌呤二核苷酸的生物发光方法]

[Bioluminescent method of determining picomolar amounts of nicotinamide-adenine dinucleotide using an immobilized extract of the luminescent bacterium Beneckea harveyi].

作者信息

Lebedeva O V, Ugarova N N, Deĭko T V, Raĭnina E I, Makhlis T A

出版信息

Prikl Biokhim Mikrobiol. 1985 Jan-Feb;21(1):114-21.

Abstract

The luminous bacteria Beneckea Harveyi were immobilized on BrCN-sepharose and cellulose films activated with cyanuric chloride. Preparations with high luciferase and FMN-reductase activities were obtained, which showed no background luminescence without NADH being added. The storage conditions for the preparations obtained were optimized, and their kinetic parameters and thermostability were studied. Standard curves for NADH determining within the concentration range 1 nM-1 microM were plotted with the detection level of 1 picomol NADH. The preparations are very promising for bioluminescent assay due to their high activity, simple production, high stability during storage and a possibility for the repeated use.

摘要

哈维氏贝内克氏发光菌被固定在溴化氰活化的琼脂糖凝胶和纤维素膜上。获得了具有高荧光素酶和FMN还原酶活性的制剂，在不添加NADH的情况下无背景发光。对所得制剂的储存条件进行了优化，并研究了其动力学参数和热稳定性。绘制了浓度范围为1 nM至1 microM的NADH测定标准曲线，NADH的检测水平为1皮摩尔。这些制剂因其高活性、生产简单、储存期间稳定性高以及可重复使用的可能性，在生物发光测定方面非常有前景。