邻苯二酚双加氧酶传感器的研究

Studies on the Catechol Dioxygenase sensor

洋葱伯克霍尔德氏菌L68的细胞裂解液经硫酸铵分级沉淀后,依次经DEAE-Sepharose Fast Flow柱层析,Hydroxyapatite柱层析和Sephadex G-150 凝胶过滤分离,提取到邻苯二酚双加氧酶.经SDS-PAGE检测,达到电泳纯.将酶反应与氧电极偶联,研究了检测水体中邻苯二酚的邻苯二酚双加氧酶传感器.以100mmol/L磷酸钠标准缓冲液(pH7.24)作为该酶传感器的的工作介质,在25℃条件下测得传感器对邻苯二酚标准液的工作曲线线性范围为0.2～3mmol/L,检测限为0.05mmol/L,响应时间为1min,平均回收率达到1.03%.

Catechol dioxygenase was extracted after the ammonium sulfate grade precipitating of Burkholderia cepacia L68 cell splitting liquor through in order of DEAE-Sepharose Fast Flow, Hydroxyapatite column chromatography and Sephadex G-150 gel filtration separation. Through SDS-PAGE detection reached electrophoretic purity. The catechol dioxygenase sensor for detecting the catachol in waters was studied with combination of enzyme reaction and oxygenic electrode. Using 100mmol/L sodium phosphate standard buffer liquor （pH7.24） as the working medium of the sensor, the work linearity range of the sensor on the catechols standard liquor was detected to be 0.2-3mmol/L, under the conditon of 25℃, the limit of detection was 0.05mmol/L, the response time was lmin and the overage recovery rate was 1.03%.