基于固定化解朊假丝酵母脲酶的电位型生物传感器测定牛奶中的尿素。
Potentiometric biosensor for determination of urea in milk using immobilized Arthrobacter creatinolyticus urease.
机构信息
Department of Biotechnology, Central Leather Research Institute, Adyar, Chennai, Tamil Nadu 600020, India.
Nuclear Agriculture and Biotechnology Division, Bhabha Atomic Research Centre, Trombay, Mumbai 400 085, India.
出版信息
Mater Sci Eng C Mater Biol Appl. 2015 Apr;49:786-792. doi: 10.1016/j.msec.2015.01.048. Epub 2015 Jan 12.
The extracellular urease from Arthrobacter creatinolyticus was partially purified by ammonium sulfate precipitation and immobilized on PAN [poly(acrylonitrile-methylmethacrylate-sodium vinylsulfonate)] membrane. The urease immobilized PAN membrane exhibited an activity of 97.92U/cm(2) under the optimum conditions of 1.0% enzyme concentration, 15% glutaraldehyde, 24h immobilization time and temperature of 4°C. The changes in surface morphology of the membrane after immobilization were studied by SEM and ATR-FTIR analysis. Immobilized membrane was associated with potentiometric electrode for calibration of biosensor and the results showed a linear response for wide range of urea concentration from 1 to 100mM. The immobilized urease had good storage stability for a period of 70days at 4°C and could be effectively reused for 13cycles. Urease immobilized membrane was also employed in analysis of urea spiked milk samples.
解朊弧菌胞外脲酶经硫酸铵沉淀部分纯化后，固定在 PAN [聚（丙烯腈-甲基丙烯酸甲酯-磺酸钠乙烯基）]膜上。在 1.0%酶浓度、15%戊二醛、24 小时固定时间和 4°C温度的最佳条件下，固定在 PAN 膜上的脲酶的活性为 97.92U/cm(2)。通过 SEM 和 ATR-FTIR 分析研究了固定化前后膜表面形貌的变化。将固定化膜与电位电极偶联，用于生物传感器的校准，结果表明，该生物传感器对 1 至 100mM 宽浓度范围内的尿素具有线性响应。固定化脲酶在 4°C 下储存 70 天具有良好的储存稳定性，可有效重复使用 13 次。固定化脲酶膜还用于分析加标牛奶样品中的尿素。
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