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本文报道运用氯化硬脂酰三甲基铵和花生酸甲酯的混合物(氯仿配制)为成膜材料,对脲酶和乙醇脱氢酶(pH7.0的 HEPES 缓冲液配制)进行吸附固定.实验在圆形 LB 膜槽上进行,先将脂溶液扩展在水面上,约10min 后,压缩单层膜至一定的初始表面压,再将单层膜转移至酶溶液表面吸附1h.吸附完毕,压缩脂-酶膜至约28 mN/m 表面压处,用石英片拉二层膜,利用分光光度计测定酶的活性.脲酶在分解尿素及乙醇脱氢酶在催化乙醇脱氢反应的过程中,辅酶烟酰胺腺嘌呤二核苷酸可被氧化或还原,从而其在340nm 处的光吸收发生变化,据此原理可测酶的活性.活性的基片上每平方厘米膜所含国际单位量表示.测活性前将基片在pH7.0的缓冲液中强烈晃动约20s 以洗去过多吸附的蛋白质.测定结果见表1.
This paper reports the use of a mixture of stearyl trimethyl ammonium chloride and methyl arachidate (chloroform preparation) as film-forming material, the urease and alcohol dehydrogenase (pH7.0 HEPES buffer preparation) adsorption fixed experiment In a circular LB membrane tank, the first lipid solution extended on the water, about 10min, the monolayer film compression to a certain initial surface pressure, and then the monolayer film was transferred to the surface of the enzyme solution adsorption 1h adsorption is completed, compressed Lipid-enzyme membrane to a surface pressure of about 28 mN / m, using a quartz film to pull the two films, using the spectrophotometer to measure the activity of urease in the decomposition of urea and alcohol dehydrogenase in the catalytic ethanol dehydrogenation reaction, The coenzyme nicotinamide adenine dinucleotide can be oxidized or reduced such that its light absorption at 340 nm changes, whereby the activity of the enzyme can be measured, expressed as the number of international units per square centimeter of film on the active substrate. Before the activity test, the substrate was shaken vigorously for about 20s in pH7.0 buffer to wash away excess adsorbed protein.