The fluorescence emission of the apo-glucose oxidase from Aspergillus niger as probe to estimate glucose concentrations
The fluorescence emission of the apo-glucose oxidase from Aspergillus niger as probe to estimate glucose concentrations
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DOI：
10.1006/bbrc.1999.1330
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发表时间：
1999-09-24
影响因子：
3.1
通讯作者：
Lakowicz, JR
中科院分区：
文献类型：
--
作者：
D'Auria, S;Herman, P;Lakowicz, JR
我们开发了一种新的方法，葡萄糖传感使用一种非活性形式的葡萄糖氧化酶从尼日尔曲霉。通过去除FAD辅因子使葡萄糖氧化酶失活。所得到的猿-葡萄糖氧化酶仍然结合葡萄糖，如从其内在色氨酸荧光的减少所观察到的。发现8-苯胺基-1-萘磺酸（ANS）自发地结合到ape-葡萄糖氧化酶，如从ANS荧光的增强所看到的。结合ANS的稳态强度在结合葡萄糖后降低25%，结合ANS的平均寿命降低约40%。这些光谱变化发生在10至20 mM葡萄糖的中点，这与空穴葡萄糖氧化酶的KD相当。这些结果表明，ape-葡萄糖氧化酶可用作可逆的非消耗性葡萄糖传感器。(C)北京：科学出版社.
We developed a new method of glucose sensing using an inactive form of glucose oxidase from Aspergillus niger. Glucose oxidase was rendered inactive by removal of the FAD cofactor. The resulting ape-glucose oxidase still binds glucose as observed from a decrease in its intrinsic tryptophan fluorescence. 8-Anilino-1-naphthalenesulfonic acid (ANS) was found to bind spontaneously to ape-glucose oxidase as seen from an enhancement of the ANS fluorescence. The steady state intensity of the bound ANS decreased 25% upon binding of glucose, and the mean lifetime of the bound ANS decreased about 40%. These spectral changes occurred with a midpoint from 10 to 20 mM glucose, which is comparable to the KD of hole-glucose oxidase. These results suggest that ape-glucose oxidase can be used as a reversible nonconsuming sensor for glucose. (C) 1999 Academic Press.