DETECTION OF HEAVY-METAL SALTS WITH BIOSENSORS BUILT WITH AN OXYGEN-ELECTRODE COUPLED TO VARIOUS IMMOBILIZED OXIDASES AND DEHYDROGENASES
DETECTION OF HEAVY-METAL SALTS WITH BIOSENSORS BUILT WITH AN OXYGEN-ELECTRODE COUPLED TO VARIOUS IMMOBILIZED OXIDASES AND DEHYDROGENASES
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DOI：
10.1016/0956-5663(93)85030-r
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发表时间：
1993-01-01
影响因子：
12.6
通讯作者：
BURSTEIN, C
中科院分区：
文献类型：
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作者：
GAYET, JC;HAOUZ, A;BURSTEIN, C
固定化氧化酶结合在亲和膜表面，并安装在氧电极上。这些生物传感器用于重金属盐的测量。固定化酶后，观察了一级失活动力学。与明胶基质中酶的网状结构相比，表面固定化增加了10倍的灵敏度。固定后，用20 μ m HgCl2对l -甘油磷酸氧化酶和50 nm丙酮酸氧化酶进行50%失活。HgCl2治疗后活性恢复是可行的，但既不完全也不可重复。为了重复利用该生物传感器，将兔肌肉溶液中的l -乳酸脱氢酶（LDH）与固定化的l -乳酸氧化酶（对重金属盐不敏感）偶联。每次测量后更换LDH（特别便宜）。磷酸盐缓冲液中HgCl2的I50为1 muM， AgNO3的I50为0.1 muM；其他重金属盐在500 muM以下无抑制作用。在Tris缓冲液中，CdCl2和ZnCl2的I50为10 muM， Pb-acetate为50 muM， CuSO4为250 muM。使用不同的酶和缓冲液可以测量特定的重金属盐。
Immobilized oxidases were bound on the surface of an affinity membrane and mounted on an oxygen electrode. These biosensors were used for heavy metal salt measurements. After immobilization of the enzymes, first order kinetics of inactivation were observed. Surface immobilization increases the sensitivity by a factor of 10, compared to reticulation of the enzyme in a gelatin matrix.After immobilization, 50% inactivation was observed with 20 muM HgCl2 for L-glycerophosphate oxidase and 50 nm for pyruvate oxidase. Restoration of activity after HgCl2 treatment is feasible, but neither complete nor reproducible. To reuse the biosensor, L-lactate dehydrogenase (LDH) from rabbit muscle in solution was coupled to immobilized L-lactate oxidase (insensitive to heavy metal salts). LDH (particularly inexpensive) was replaced after each measurement. The I50 in phosphate buffer was 1 muM for HgCl2 and 0.1 muM for AgNO3; with other heavy metal salts, no inhibition was observed below 500 muM. In Tris buffer, the I50 was 10 muM for CdCl2 and ZnCl2, 50 mum for Pb-acetate and 250 muM for CuSO4.The use of different enzymes and buffers may allow measurement of specific heavy metal salts.