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利用来自纤细杆菌X的海藻糖稳定冻干甲醇脱氢酶检测甲醇。

The use of trehalose-stabilized lyophilized methanol dehydrogenase from Hyphomicrobium X for the detection of methanol.

作者信息

Argall M E, Smith G D

机构信息

Division of Biochemistry and Molecular Biology, School of Life Sciences, Australian National University, Canberra.

出版信息

Biochem Mol Biol Int. 1993 Jul;30(3):491-7.

PMID:8401307
Abstract

The enzyme methanol dehydrogenase (EC 1.1.99.8) from Hyphomicrobium X was used in an attempt to develop a rapid colorimetric test for methanol. The enzyme was stabilized for storage by lyophilization in the presence of the disaccharide trehalose. It was found that the enzyme retained significantly greater activity in the dried state with trehalose than without. The enzyme was partially purified by ammonium sulphate fractionation, after which it was found to be more stable in solution at pH 9 than at pH 7. A procedure is given which involves mixing a defined amount of enzyme with the methanol-containing water together with phenazine methosulphate (PMS), 2-6-dichlorophenol-indophenol (DCPIP) and cyanide, and observing the resultant colour change from blue to yellow if methanol is present. The sensitivity of the procedure is such that 9 mg L-1 of methanol can be readily detected.

摘要

来自X型生丝微菌的甲醇脱氢酶(EC 1.1.99.8)被用于尝试开发一种快速比色法检测甲醇。通过在二糖海藻糖存在下冻干来稳定该酶以便储存。发现该酶在有海藻糖的干燥状态下比没有时保留显著更高的活性。通过硫酸铵分级分离对该酶进行部分纯化,之后发现它在pH 9的溶液中比在pH 7时更稳定。给出了一个程序,该程序包括将规定量的酶与含甲醇的水以及硫酸吩嗪甲酯(PMS)、2,6-二氯酚靛酚(DCPIP)和氰化物混合,如果存在甲醇则观察由此产生的从蓝色到黄色的颜色变化。该程序的灵敏度使得9 mg L-1的甲醇能够很容易被检测到。

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